{"paper_id":"33ebba1c-e85e-48d7-b927-406ba4f8dbe4","body_text":"Dexmedetomidine promotes NREM sleep by depressing the neuronal activity of OXT neurons in the PVN | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Dexmedetomidine promotes NREM sleep by depressing the neuronal activity of OXT neurons in the PVN Ying Zhang, Jiaxin Li, Wei Wang, Daming Wang, Junli Ding, Liecheng Wang, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4207174/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 30 Jul, 2024 Read the published version in Neurochemical Research → Version 1 posted 7 You are reading this latest preprint version Abstract This study aimed to investigate the mechanism of Dexmedetomidine (DEX) on sleep homeostasis. We will focus on the effect of DEX on the activity and the signal transmission of oxytocin-expressing (Oxytocin, OXT) neurons in the paraventricular nucleus (PVN) of the hypothalamus (PVN OXT ) in the regulation of sleep-wakefulness cycle. In this study, the guide cannula was implanted to microinject DEX into PVN. PVN OXT neurons were specifically activated by the chemogenetic method. Differences in sleep-wakefulness states were monitored by electroencephalogram (EEG)/ electromyogram (EMG) recording. In addition, the effect of DEX on the electrophysiological activity of PVN OXT neurons was examined by whole-cell patch-clamp technique. EEG/EMG results showed that microinjection of DEX in PVN significantly increased the duration of nonrapid eye movement (NREM) sleep in mice. Chemogenetic activation of PVN OXT neurons after internal cannula injection of DEX to PVN increased the amount of wake. Electrophysiological results show that DEX could inhibit the frequency of action potential (AP) and the spontaneous excitatory postsynaptic current (sEPSC) in PVN OXT neurons through α 2 -adrenoceptors. DEX maintained sleep homeostasis by inhibiting excitatory synaptic signaling in PVN OXT neurons through α 2 -adrenoceptors. DEX OXT neurons PVN EEG/EMG recording patch clamp Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 1 Introduction Dexmedetomidine (DEX) is a highly specific α 2 -adrenoreceptor agonist, with a 1620 times affinity of binding to α 1 adrenoreceptors [ 1 ]. α 2 -adrenoceptors are a class of heterotrimeric guanine nucleotide-binding protein receptors (GPCR) [ 2 ], which play a wide role in regulating physiological functions, including heart rate, blood pressure, blood glucose, insulin homeostasis, and analgesia. They are involved in presynaptic feedback inhibition mediating the release of neurotransmitters from the terminal of norepinephrine synapses [ 3 – 5 ]. As an anesthetic drug, DEX has the characteristics of sedation and analgesia with few side effects and a wide application range[ 6 – 12 ]. The specific role of DEX in postoperative sleep and its related mechanisms are poorly understood. As a sedative-hypnotic drug, DEX has a certain impact on the sleep-wake “switches” [ 13 ]. Intraperitoneal injection of DEX can increase the c-Fos expression in the overlapping areas of the preoptic area (POA) and diaphragm of the hypothalamus [ 14 ], ventral tegmental area (VTA) [ 15 ], and area postrema (AP*) [ 16 ] and decrease the c-Fos expression in the supraoptic nucleus (SON) [ 17 ], primary motor cortex (M1), and secondary motor cortex (M2) [ 18 ], and these brain regions are associated with the modulation of sleep-wakefulness [ 19 ]. Studies have shown that DEX can increase the calcium signal intensity of dopamine (DA) neurons in the VTA (VTA DA ). DEX can increase the excitability of VTA DA neurons through α adrenoreceptors. It also increases the concentration of DA neurotransmitters in the medial prefrontal cortex (MPFC) and nucleus accumbens (NAc), an effect that may be related to the sedative arousal properties of DEX [ 15 ]. The activation of VTA DA neurons can induce the state change from an unconscious anesthetic to wakefulness, triggering a strong awakening-promoting effect [ 20 ], which may be the mechanism of DEX-induced rapid awakening after sedation. Recently, it was found that vesicular glutamate transporter 2 (Vglut2), oxytocin (OXT), prodynorphin (PDYN), corticotropin-releasing hormone (CRH), and vasopressin (AVP) neurons in the paraventricular nucleus of the hypothalamus (PVN) could induce different levels of arousal through chemogenetic activation [ 21 ]. PVN is composed of three types of neurons: the large cells (Magno), the small cells (Parvocellular), and the long projection neurons, which express OXT, PDYN, CRH, et.al. Almost one-third of neurons in PVN are expressing OXT (PVN OXT ) mainly distributed in the middle part of PVN. [ 22 ]. AVP and OXT neurons are involved in the regulation of physiology and behavior, including stress response and social behavior. Sleep-wakefulness has an essential attribution to these adaptive behaviors. Our previous study showed that DEX could depress the c-Fos expression of AVP neurons in PVN and reduce the concentration of AVP in serum [ 23 ]. It has been found that photogenetic and chemogenetic activation of PVN AVP neurons significantly increases the amount of wake in mice [ 21 , 24 ]. PVN OXT neurons are an important component in the regulation of sleep and awakening [ 25 ]. Studies have shown that chemogenetic activation of PVN OXT neurons could increase the amount of wake and decrease the amount of NREM sleep and rapid eye movement (REM) sleep [ 21 ]. Here, we aim to investigate the roles of PVN OXT neurons in the regulation of sleep homeostasis by DEX. We used the chemogenetic methods to activate PVN OXT neurons after canula administration of DEX to PVN, we found that activation of PVN OXT neurons could counteract the increase of NREM sleep time induced by DEX. In addition, we found that DEX inhibited the frequency of action potential (AP) and spontaneous excitatory postsynaptic current (sEPSC) of PVN OXT neurons through α 2 -adrenoceptors. It is suggested that DEX might maintain sleep homeostasis by inhibiting excitatory synaptic signaling in PVN OXT neurons. 2 Materials and Methods 2.1 Mice The animal experimental procedures underwent rigorous scrutiny and approval by the Laboratory Animal Management Office of Laboratory Animals at Anhui Medical University, adhering strictly to the pertinent ethical directives established by the Institutional Animal Care Unit Committee of Anhui Medical University, under project identifier LLSC20190763. OXT-IRES-Cre mice (stock No. 024234; Jackson Laboratory, Bar Harbor, ME) and Ai9 mice (stock No. 007909; Jackson Laboratory, Bar Harbor, ME) were generously provided by Professor Tian Xue from the University of Science and Technology of China. Housing conditions for all mice were maintained under a 12-hour light-dark cycle (lights on at 08:00, illuminance ranging from 150–200 lux), with ad libitum access to food and water, and a room temperature (RT) set at 25°C. 2.2 Surgery and virus injection Mice were deeply anesthetized with 50 mg/kg sodium pentobarbital and positioned in a stereotaxic frame (RWD Life Science, Shenzhen, China) with sterilized instruments. Utilizing coordinates from the mouse brain atlas[ 26 ], a microinjection needle delivered 200 nL of rAAV-EF1α-DIO-hM3D(Gq)-mCherry-WPREs (≥ 5.00E + 12 viral particles/mL, Brainvta, Wuhan, China) bilaterally into the PVN at a rate of 40 nL/min (M/L: ± 0.30 mm, A/P: -0.6 mm, D/V: -4.6). The guide cannula (RWD Life Science, Shenzhen, China) was implanted in the PVN with sagittal plane tilting 5°(M/L: 1.0 mm, A/P: -0.6 mm, D/V: -4.6) and secured with light-curable resin (BJBCJB, Sino-Dentex, Jilin, China). The length of internal cannulas (RWD Life Science, Shenzhen, China), is equal to guide cannulas. EEG electrodes were inserted into the skull (+ 1.5 mm Bregma, -1.5 mm midline – first recording electrode; +1.5 mm Lambda, + 1.5 mm midline – second recording electrode), while EMG electrodes were placed in neck musculature. All electrodes were affixed with light-cured resin and quick self-curing acrylic resin (SC01, Yamahahi Dental, Japan). Surgical sites were sutured and disinfected. Post-surgery, mice were housed individually. 2.3 EEG/EMG recordings and analysis The animals were housed individually in an EEG/EMG recording cage under a 12-hour light:12-hour dark cycle (with light onset at 08:00 and offset at 20:00) for 3 days of acclimation. Electrical signals were recorded at 1 kHz, amplified with the MP150 system (BIOPAC, California, USA), and digitized using Acknowledge 4.2 software (BIOPAC, California, USA). Automated analysis categorized EEG/EMG signals into Wake, NREM sleep, and REM sleep states every 4 seconds using the Lunion Stage AI engine (LunionData, Shanghai, China), with manual verification and correction. EEG signals were filtered between 0.3–35 Hz and EMG signals between 10–200 Hz. 2.4 Electrophysiology 2.4.1 Slice preparation OXT::Ai9 mice (8–12 weeks old) were anesthetized with 1% isoflurane (RWD Life Science, Shenzhen, China) and decapitated. The brain was swiftly excised and immersed in a freezing oxygenated (95% O 2 and 5% CO 2 ) cutting solution containing N-methyl-D-glucamine, 1-deoxy-1-(methylamine)-D-sorbito, glucamine (NMDG) cutting solution (in mM: 92 NMDG, 1.2 potassium chloride (KCl), 1.2 potassium dihydrogen phosphate (KH 2 PO 4 ), 30 sodium bicarbonate (NaHCO 3 ), 25 D-glucose, 20 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES), 5 L-ascorbic acid, 3 sodium pyruvate, 2 thiourea, 10 magnesium sulfate (MgSO 4 ) and 0.5 calcium chloride (CaCl 2 ); the pH adjusted to 7.2 ± 0.1 with hydrogen chloride (HCl), the osmolarity adjusted to 310 ± 5 mOsmol/L with NMDG). Horizontal brain slices (300 µm) were then obtained using a vibrating microtome (VT1200S, Leica, Germany). These slices were transferred to oxygenated (95% O 2 and 5% CO 2 ) aCSF (mM: 125 NaCl, 1.25 KCl, 2 CaCl 2 , 1 MgCl 2 , 1.25 KH 2 PO 4 , 25 NaHCO 3 , 0.4 Na-pyruvate, 0.08 L-ascorbic acid, 25 D-glucose), and allowed incubating for 40–45 minutes at 36.6 ℃ before recording. 2.4.2 Whole-cell recordings Whole-cell recordings were performed at RT in oxygenated (95% O 2 and 5% CO 2 ) aCSF solution with constant perfusion at a rate of 1–2 mL/min. For action potential (AP) recording, the electrodes were filled with a potassium-based internal solution (in mM: 125 K-gluconate, 10 KCl, 10 HEPES, 0.1EGTA, 5 ATP-Mg, 0.5GTP-Na; the pH adjusted to 7.2 ± 0.1 with potassium hydroxide (KOH); the osmolarity adjusted to 310 ± 5 mOsmol/kg with K-gluconate). For sEPSCs recording, the electrodes were filled with a CsMethane sulfonate-based internal solution containing (in mM: 135 CsMethane sulfonate, 10 cesium chloride (CsCl), 10 HEPES, 1EGTA, 5 ATP-Mg, 0.5 GTP-Na, the pH was adjusted to 7.2 ± 0.1 using cesium hydroxide (CsOH), and the osmolarity was adjusted to 310 ± 5 mOsmol/L using CsMethane sulfonate). The aCSF with 50 µM picrotoxin (PTX, HY-101391, MedChemExpress, USA) was recorded sEPSCs. For sIPSCs recording, the electrodes were filled with a cesium-based internal solution containing (in mM: 140 CsCl, 10 HEPES, 1EGTA, 5 ATP-Mg, 0.5 GTP-Na; the pH was adjusted to 7.2 ± 0.1 using CsOH, and the osmolarity was adjusted to 310 ± 5 mOsmol/L using CsCl). The aCSF with 10 µM 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, ab120017, Abcam, UK) and 20 µM 5-phospho-D-norvaline (D-AP5, ab120003, Abcam, UK) were recorded sIPSCs. sEPSCs were recorded in the presence of the GABA A receptor antagonist Picrotoxin (PTX) and sIPSCs were recorded in the presence of AMPA and NMDA receptor antagonists CNQX and D-AP5 in the aCSF. The neurons were held at -70 mV. The resistance of the patch pipette was 5–7 MΩ. A 40× water microscope (TH4-200 and U-RFL-T microscope, Olympus, Japan) was used to visualize the cells. Data were collected by using an Axopatch 700B patch clamp amplifier, Digidatal 1550B data acquisition system, and pClamp10.7 software (Molecular Devices, USA). Data were sampled at 20 kHz, and low-pass filtered with a cutoff frequency of 1 kHz in the current recording. Clampfit 10.7 (Molecular Devices, USA) was used to analyze the electrophysiological properties of the cells. If the hold current of the patch neuron exceeds 30 pA during whole-cell recording or the change of Access resistance (Ra) exceeds 20% during recording, statistics will not be included in the statistical analysis. Igor Pro (WaveMetrics, USA) was used to make statistical charts and representative charts. 2.5 Drug administration In the EEG/EMG recording, OXT-hM3D(Gq)-mCherry mice were recorded from Zeitgeber time (ZT) 14 to ZT0, after the cannula administration of saline or 2 µM DEX to investigate the DEX-induced effect of sleep-wakefulness states (Fig. 1 b). OXT-hM3D(Gq)-mCherry mice were injected 1µL 2 µM DEX into internal cannulas at ZT14(22:00) and i.p injected saline or CNO at ZT15.5(23:30) to investigate the effect of sleep-wakefulness states in PVN OXT neurons (Fig. 2 c) [ 27 , 28 ]. Electrophysiological characteristics of PVN OXT neurons of APs were captured using the whole-cell patch-clamp method in brain slices immersed in the artificial cerebrospinal fluid (aCSF) with or without 2 µM DEX. RS79948, an α 2 -adrenergic receptor antagonist, was utilized to block adrenergic signaling [ 29 ]. Following a 2-minute stabilization, PVN OXT neurons were microinjected with 1 mM RS79948 for 30 seconds, followed by a 3-minute recording period. Subsequently, neurons received a combined microinjection of 1 mM RS79948 and 2 µM DEX for 30 seconds, with a further 3-minute recording. (Fig. 4 a). To assess the impact of DEX on neuronal signal transmission, sEPSCs and spontaneous inhibitory postsynaptic currents (sIPSCs) were recorded. Initially, neurons were stabilized for 2 minutes, then microinjected with aCSF for 30 seconds, and data were recorded for 2 minutes. This was followed by 30-second microinjection of 2 µM DEX, with a 2-minute recording period (Fig. 5 a). To determine the factors of the observed sEPSCs reduction, neurons were stabilized for 2 minutes, microinjected with 1 mM RS79948 for 30 seconds, and recorded for 1 minute, followed by a co-injection of 1 mM RS79948 and 2 µM DEX for 30 seconds, with a final 1-minute recording period (Fig. 5 b). 2.6 Statistical Analysis Data are presented as mean ± standard error of the mean (SEM). The normality of data was assessed using Shapiro-Wilk analysis prior to t-tests. Statistical comparisons between two groups were conducted using paired or unpaired Student's t-tests, Wilcoxon matched-pairs signed rank test, or Mann-Whitney test as appropriate. Multiple parameter comparisons were performed using two-way repeated-measures ANOVA, followed by Bonferroni's multiple comparison test. Prism 10.0 software (GraphPad Software, San Diego, CA, USA) was utilized for all statistical analyses. P < 0.05 was considered statistically significant. 3 Result 3.1 DEX increased NREM sleep time and decreased the average duration of wakefulness. For further exploration of the mechanism of DEX in the sleep-wakefulness cycle, EEG/EMG was divided into three states: NREM sleep, REM sleep, and wake. A guide cannula for drug administration was implanted in the PVN. The mice were cannula injected 1µL saline or 2 µM DEX into PVN with a speed of 1µL/min (Fig. 1 a). We used the EEG/EMG recording to analyze the sleep-wakefulness phases in mice between the saline group and the DEX group (Fig. 1 c-d). The results showed that the amount of wake during ZT14-18 was decreased (Fig. 1 e), the amount of NREM sleep during ZT14-18 was increased (Fig. 1 f), and the amount of REM sleep during ZT14-18 had no differences (Fig. 1 g) in the DEX group, compared with the saline group., EEG Power spectrum density (PSD) in the DEX group decreased significantly from 1.0-3.9Hz within 4 hours, compared with the saline group (Fig. 1 h). DEX resulted in a decrease in the mean duration of wake (Fig. 1 i), but the number of sleep transitions had no differences (Fig. 1 j). 3.2 Chemogenetic activation of PVN OXT neurons counteracted DEX-induced NREM sleep and increased arousal. We injected rAAV-EF1α-DIO-hM3D(Gq)-mCherry-WPREs into PVN to infect OXT neurons (Fig. 2 a). Patch clamp recording was used to certify the effectiveness of the virus. The OXT-hM3D(Gq)-expressing neurons were puff with the aCSF and 5µm clozapine-N-oxide (CNO, Brainvta, Wuhan, China) for 5 minutes respectively, indicating that neurons could be successfully activated by CNO (Fig. 2 b). The results of i.p. injection of saline or 2 mg/kg CNO into ZT2 showed that activation of PVN OXT neurons in the CNO group could increase the amount of wake and decrease NREM sleep in 1 hour, while REM sleep time had no differences (Fig. 2 d-f). EEG/EMG was recorded for 10 hours from ZT14 (Fig. 2 g, h). After 30 minutes of CNO injection, the time of three sleep states of 0.5, 1, and 1.5 hours were counted. Compared with the saline group, after chemogenetic inhibition of PVN OXT neurons, the amount of wake during ZT16-17 in the CNO group was increased, while the amount of NREM time was decreased and the amount of REM had no differences (Fig. 2 i-m). In conclusion, the DEX-induced increase in NREM sleep was counteracted by the activation of PVN OXT neurons. 3.3 DEX inhibited the AP frequency of PVN OXT neurons. To further investigate the mechanism of the effect of DEX on PVN OXT neurons on sleep homeostasis, we prepared brain slices containing PVN from OXT::Ai9 mice during ZT13 and ZT14. During electrophysiological recording, fluorescent images of recorded PVN OXT neurons were collected (Fig. 3 a). We examined the firing properties of PVN OXT neurons before and after drug administration, such as the AP duration were measured at 50% repolarization (APD 50 ), AP amplitude, spike max, afterhyperpolarization (AHP) amplitudes, thresholds, resting cell membrane potential (RMP), membrane input resistance (R in ) (Fig. 3 b). The results showed that the AP frequency of OXT neurons in the DEX group was significantly reduced, compared to the aCSF group (Fig. 3 c). However, there were no significant differences in APD 50 , AP amplitude, spike max, AHP, threshold, RMP, and R in between the aCSF group and the DEX group (Fig. 3 d-j). When the neurons received stimulation from − 40 pA to 0 pA, the DEX group showed a tendency to decrease the potential difference, compared with the aCSF group (Fig. 3 k). DEX might reduce the rate of ion influx. Further analysis of the changes in membrane potential and resistance will help to reveal the finer regulatory mechanism of DEX on the function of OXT neurons. 3.4 DEX decreased the AP and sEPSC frequencies of PVN OXT neurons through α 2 -adrenoceptors. As mentioned above, DEX is a highly potent α 2 -adrenoceptors agonist. We recorded the effect of blocking the receptor-mediated signaling pathway on the electrophysiological characteristics of PVN OXT neurons by patch-clamp experiments. The results showed that the AP frequency, APD 50 , AP amplitude, spike max, AHP amplitude, threshold, RMP, and R in have not changed in the RS79948 + DEX group, compared with the RS79948 group (Fig. 4 b-j). This suggests that DEX might inhibited the AP frequency in PVN OXT neurons through α 2 -adrenoceptors. Subsequently, sEPSCs and sIPSCs were recorded to explore whether DEX could influence the neuronal signal transmission of PVN OXT neurons. We observed a significant decrease in the frequency of sEPSC in the PVN OXT neurons between the aCSF group and the DEX group (Fig. 5 c), and no significant difference between the mean amplitude of sEPSC (Fig. 5 d). DEX did not show significant effects on both the frequency and amplitude of sIPSC (Fig. 5 e, f). Last, we puffed RS79948 to both the aCSF group and the DEX group and recorded sEPSCs and sIPSCs in PVN OXT neurons. We found that RS79948 blocked the decreased frequency of sEPSC induced by DEX, with no difference in the amplitude of sEPSC (Fig. 5 g, h). Combined with the above results, our study shows that DEX has a significant effect on modulating AP and excitatory synaptic transmission in PVN OXT neurons through α 2 -adrenoceptors. 4 Discussion Clinical studies show that severe sleep disorders and delirium often occur after surgery [ 30 ]. Patients exhibit sleep deprivation, sleep fragmentation, and a reduction in the duration of slow-wave sleep and REM sleep [ 31 ]. The most significant features of postoperative delirium include cognitive deficits, dysfunction of circadian rhythm, and emotional disorders. Studies have shown that the DEX has characteristics of low side effects, which could improve postoperative sleep quality, and reduce the incidence of postoperative delirium after surgery [ 32 , 33 ]. Disturbance of sleep structure is a serious problem for postoperative patients, which may lead to prolonged postoperative recovery time [ 34 ] and increase the incidence of cardiovascular diseases, cognitive dysfunctions [ 35 ], and impaired immune functions [ 36 ]. Therefore, we hope to find out the mechanism of DEX on the modulation of sleep homeostasis, further reducing the incidence of delirium. Studies showed that oral administration of DEX can increase the duration of NREM sleep within 6 hours, and decrease the duration of REM sleep within 6 hours[ 13 ]. In a sleep deprivation experiment, the polysomnography results show that the delta wave spectrum in NREM sleep would increase during the recovery period after sleep deprivation [ 37 ]. DEX-induced sedation is similar to deep restorative sleep after sleep deprivation. Unlike the classical hypnotic drug diazepam, oral administration of DEX could increase the power of the delta spectrum in NREM sleep EEG, especially at high-frequency delta waves of 1.75–3.25 Hz, while the power of low-frequency delta waves of 0.5-1.0 Hz was reduced. The same phenomenon appeared in our study, the cannula injection of DEX to PVN could increase the amount of NREM sleep time and decrease the mean duration of wakefulness, and DEX decreased the delta spectrum at 1.0-3.9Hz. If subsequent studies can find the effect of DEX on different delta wave frequencies, it will be helpful to further analyze the regulatory effect of DEX on NREM sleep [ 13 ]. OXT-expressing neurons are predominantly located in the PVN and supraoptic nucleus (SON) of the mammalian hypothalamus. OXT plays a crucial role in modulates responses to stress, fear, and social behaviors[ 38 – 42 ]. Little research has investigated the influence of OXT on sleep-wakefulness. Interestingly, OXT might have a dual mechanism of action in dependence of the physiological state on sleep regulation. Long-term intranasal administration of OXT reduced sleep latencies and increased the sleep efficiency and percentage of REM sleep episodes in human [ 43 ]. A study in mice showed that acute lateral ventricle (ICV) infusion of OXT delayed sleep onset latency, which resulted in a transient reduction of NREM sleep and REM sleep, and augmented high-frequency activity in NREM sleep [ 44 ]. Optogenetic activation of OXT neurons in PVN can promote arousal [ 45 ]. The same phenomenon was found in our study, the chemogenetic activation of PVN OXT neurons could increase the amount of wake. We found that the activation of wakeful-promoting PVN OXT neurons through the chemogenetic method could decrease the one-hour amount of NREM sleep induced by DEX. DEX-induced sedation has the property of deep restorative sleep after sleep deprivation [ 14 ] and is easy to wake up [ 46 ]. Whether this process was modulated by working on OXT neurons needs further studies. Different firing patterns of neurons dictate the amount and timing of neuropeptide release, and the activity of the network affects the neurons' integrative properties. The previous study indicates DEX can increase or decrease delta and theta wave percentage through chemogenetic inhibition or activation of VTA DA neurons. DEX increases the firing rate of VTA DA neurons through α 2 adrenoceptors [ 15 ]. DEX is widely recognized for its sedative properties and is predominantly believed to exert its effects by engaging the norepinephrine system. Extensive research indicates that DEX interacts with α 2 adrenoceptors located on the norepinephrine-producing neurons of the LC. This interaction triggers a hyperpolarization of the neuronal cell membranes via a Gi protein-mediated pathway. As a result, the release of the stimulating neurotransmitter norepinephrine is diminished across the brain, culminating in a state of sedation[ 47 ]. Here, we examined the influence of DEX on the membrane characteristics and synaptic properties of PVN OXT neurons. We found DEX decreases the frequency of AP in PVN OXT neurons. The chemogenetic activation of PVN Vglut2 neurons can increase arousal indicate glutamate signaling pathway participates in modulating the sleep-wakefulness cycle [ 21 ], and in our study, we also found DEX could decrease the frequency of sEPSC of PVN OXT neurons. Therefore, DEX might reduce the rate of ion influx by bounding α 2 adrenoceptors, resulting in decreased excitability of the PVN OXT neurons. In conclusion, these findings highlighted DEX increased NREM sleep by inhibiting PVN OXT neurons, which promoted arousal and decreased the frequency of AP and sEPSC of PVN OXT neurons. The results reveal the potential role of modulation of DEX in PVN OXT neurons and provide new ideas for the treatment of postoperative delirium prevention. However, further research is still needed to resolve the molecular mechanisms of DEX and its more far-reaching effects on neuronal networks. Declarations Acknowledgments The authors thank Dandan Zang, Cheng Qian, and Guanjun Chen of the Center for Scientific Research of Anhui Medical University for their valuable help in our experiments. Author Contributions Juan Cheng and Liecheng Wang supervised and designed the work; Ying Zhang and Juan Cheng wrote and revised the manuscript; Ying Zhang and Jiaxin Li completed the patch-clamp experiment, EEG/EMG recording and gene identification. Ying Zhang, Jiaxin Li, Wei Wang, Daming Wang, and Junli Ding analyzed the experiment data and bred the mice. Data Availability Statement: Raw data will be made available on request to the corresponding author. Declaration of Conflicting Interests The author(s) declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article. Funding The author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Our work was supported by the National Natural Science Foundation of China (31800997 to Juan Cheng, 81971236, 81571293 to Liecheng Wang), the Natural Science Foundation of Universities of Anhui Province (2022AH050783 to Juan Cheng), Anhui Provincial Natural Science Foundation (2308085MC83 to Juan Cheng), Research Fund of Anhui Institute of translational medicine (2023zhyx-C71 to Daming Wang) , Basic and Clinical Cooperative Research Promotion Program of Anhui Medical University (2020xkjT027 to Junli Ding). References Weerink MAS, Struys M, Hannivoort LN, Barends CRM, Absalom AR, Colin P (2017) Clinical Pharmacokinetics and Pharmacodynamics of Dexmedetomidine. Clin Pharmacokinet 56:893–913. https://doi.org:10.1007/s40262-017-0507-7 Link R, Daunt D, Barsh G, Chruscinski A, Kobilka B (1992) Cloning of two mouse genes encoding alpha 2-adrenergic receptor subtypes and identification of a single amino acid in the mouse alpha 2-C10 homolog responsible for an interspecies variation in antagonist binding. Mol Pharmacol 42:16–27 Wang CD, Buck MA, Fraser CM (1991) Site-directed mutagenesis of alpha 2A-adrenergic receptors: identification of amino acids involved in ligand binding and receptor activation by agonists. Mol Pharmacol 40:168–179 Ramos BP, Arnsten AF (2007) Adrenergic pharmacology and cognition: focus on the prefrontal cortex. Pharmacol Ther 113:523–536 Maletic V, Eramo A, Gwin K, Offord SJ, Duffy RA (2017) The Role of Norepinephrine and Its α-Adrenergic Receptors in the Pathophysiology and Treatment of Major Depressive Disorder and Schizophrenia: A Systematic Review. Front Psychiatry 8:42. https://doi.org:10.3389/fpsyt.2017.00042 Sun S, Wang J, Bao N, Chen Y, Wang J (2017) Comparison of dexmedetomidine and fentanyl as local anesthetic adjuvants in spinal anesthesia: a systematic review and meta-analysis of randomized controlled trials. Drug Des Devel Ther 11:3413–3424. https://doi.org:10.2147/DDDT.S146092 Barends CR, Absalom A, van Minnen B, Vissink A, Visser A (2017) Dexmedetomidine versus Midazolam in Procedural Sedation. A Systematic Review of Efficacy and Safety. PLoS ONE 12:e0169525. https://doi.org:10.1371/journal.pone.0169525 Lin Y, Zhang R, Shen W, Chen Q, Zhu Y, Li J, Chi W, Gan X (2020) Dexmedetomidine versus other sedatives for non-painful pediatric examinations: A systematic review and meta-analysis of randomized controlled trials. J Clin Anesth 62:109736. https://doi.org:10.1016/j.jclinane.2020.109736 V ISSRG, W JK, VA. D MM (1988) Dexmedetomidine Diminishes Halothane Anesthetic Requirements in Rats Through a PostsynapticAlphaz Adrenergic Receptor. Anesthesiology Brown EN, Pavone KJ, Naranjo M (2018) Multimodal General Anesthesia: Theory and Practice. Anesth Analg 127:1246–1258. https://doi.org:10.1213/ane.0000000000003668 Hall JE, Uhrich TD, Barney JA, Arain SR, Ebert TJ (2000) Sedative, amnestic, and analgesic properties of small-dose dexmedetomidine infusions. Anesth Analg 90:699–705. https://doi.org:10.1097/00000539-200003000-00035 Lobo FA, Wagemakers M, Absalom AR (2016) Anaesthesia for awake craniotomy. Br J Anaesth 116:740–744. https://doi.org:10.1093/bja/aew113 Feng ZX, Dong H, Qu WM, Zhang W (2018) Oral Delivered Dexmedetomidine Promotes and Consolidates Non-rapid Eye Movement Sleep via Sleep-Wake Regulation Systems in Mice. Front Pharmacol 9:1196. https://doi.org:10.3389/fphar.2018.01196 Zhang Z, Ferretti V, Guntan I, Moro A, Steinberg EA, Ye Z, Zecharia AY, Yu X, Vyssotski AL, Brickley SG, Yustos R, Pillidge ZE, Harding EC, Wisden W, Franks NP (2015) Neuronal ensembles sufficient for recovery sleep and the sedative actions of alpha2 adrenergic agonists. Nat Neurosci 18:553–561. https://doi.org:10.1038/nn.3957 Qiu G, Wu Y, Yang Z, Li L, Zhu X, Wang Y, Sun W, Dong H, Li Y, Hu J (2020) Dexmedetomidine Activation of Dopamine Neurons in the Ventral Tegmental Area Attenuates the Depth of Sedation in Mice. Anesthesiology 133:377–392. https://doi.org:10.1097/ALN.0000000000003347 Bronzino JD, Stern WC, Leahy JP, Morgane PJ (1976) Sleep cycles in cats during chronic electrical stimulation of the area postrema and the anterior raphe. Brain Res Bull 1:235–239. https://doi.org:10.1016/0361-9230(76)90073-3 Jiang-Xie LF, Yin L, Zhao S, Prevosto V, Han BX, Dzirasa K, Wang F (2019) A Common Neuroendocrine Substrate for Diverse General Anesthetics and Sleep. Neuron 102:1053–1065e1054. https://doi.org:10.1016/j.neuron.2019.03.033 Lemke SM, Ramanathan DS, Darevksy D, Egert D, Berke JD, Ganguly K (2021) Coupling between motor cortex and striatum increases during sleep over long-term skill learning. Elife 10. https://doi.org:10.7554/eLife.64303 Li H (2023) A preliminary study of brain response region in mice after intraperitoneal injection of dexmedetomidine. Master, Anhui Medical University Taylor NE, Van Dort CJ, Kenny JD, Pei J, Guidera JA, Vlasov KY, Lee JT, Boyden ES, Brown EN, Solt K (2016) Optogenetic activation of dopamine neurons in the ventral tegmental area induces reanimation from general anesthesia. Proc Natl Acad Sci U S A 113:12826–12831. https://doi.org:10.1073/pnas.1614340113 Chen CR, Zhong YH, Jiang S, Xu W, Xiao L, Wang Z, Qu WM, Huang ZL (2021) Dysfunctions of the paraventricular hypothalamic nucleus induce hypersomnia in mice. Elife 10. https://doi.org:10.7554/eLife.69909 Xu S, Yang H, Menon V, Lemire AL, Wang L, Henry FE, Turaga SC, Sternson SM (2020) Behavioral state coding by molecularly defined paraventricular hypothalamic cell type ensembles. Science 370. https://doi.org:10.1126/science.abb2494 Yang W, Li H, Cheng Z, Lu Y, Li W, Feng J, Wang L, Cheng J (2022) Dex modulates the balance of water-electrolyte metabolism by depressing the expression of AVP in PVN. Front Pharmacol 13:919032. https://doi.org:10.3389/fphar.2022.919032 Islam MT, Rumpf F, Tsuno Y, Kodani S, Sakurai T, Matsui A, Maejima T, Mieda M (2022) Vasopressin neurons in the paraventricular hypothalamus promote wakefulness via lateral hypothalamic orexin neurons. Curr Biol 32:3871–3885e3874. https://doi.org:10.1016/j.cub.2022.07.020 Jurek B, Neumann ID (2018) The Oxytocin Receptor: From Intracellular Signaling to Behavior. Physiol Rev 98:1805–1908. https://doi.org:10.1152/physrev.00031.2017 Paxinos G, Franklin KBJ (2004) The mouse brain in stereotaxic coordinates. Elsevier Academic, Amsterdam; Boston Ge C, Chen W, Zhang L, Ai Y, Zou Y, Peng Q (2023) Chemogenetic activation of the HPC-mPFC pathway improves cognitive dysfunction in lipopolysaccharide -induced brain injury. Theranostics 13:2946–2961. https://doi.org:10.7150/thno.82889 Tang HD, Dong WY, Hu R, Huang JY, Huang ZH, Xiong W, Xue T, Liu J, Yu JM, Zhu X, Zhang Z (2022) A neural circuit for the suppression of feeding under persistent pain. Nat Metab 4:1746–1755. https://doi.org:10.1038/s42255-022-00688-5 Fan S, Cheng X, Zhang P, Wang Y, Wang L, Cheng J (2023) The α(2) Adrenoceptor Agonist and Sedative/Anaesthetic Dexmedetomidine Excites Diverse Neuronal Types in the Ventrolateral Preoptic Area of Male Mice. ASN Neuro 15:17590914231191016. https://doi.org:10.1177/17590914231191016 Su X, Meng Z-T, Wu X-H, Cui F, Li H-L, Wang D-X, Zhu X, Zhu S-N, Maze M, Ma D (2016) Dexmedetomidine for prevention of delirium in elderly patients after non-cardiac surgery: a randomised, double-blind, placebo-controlled trial. Lancet 388:1893–1902. https://doi.org:10.1016/s0140-6736(16)30580-3 Chouchou F, Khoury S, Chauny JM, Denis R, Lavigne GJ (2014) Postoperative sleep disruptions: a potential catalyst of acute pain? Sleep Med Rev 18:273–282. https://doi.org:10.1016/j.smrv.2013.07.002 Song B, Li Y, Teng X, Li X, Yang Y, Zhu J (2019) The Effect Of Intraoperative Use Of Dexmedetomidine During The Daytime Operation Vs The Nighttime Operation On Postoperative Sleep Quality And Pain Under General Anesthesia. Nat Sci Sleep 11:207–215. https://doi.org:10.2147/nss.S225041 Carr ZJ, Cios TJ, Potter KF, Swick JT (2018) Does Dexmedetomidine Ameliorate Postoperative Cognitive Dysfunction? A Brief Review of the Recent Literature. Curr Neurol Neurosci Rep 18:64. https://doi.org:10.1007/s11910-018-0873-z Gögenur I, Bisgaard T, Burgdorf S, van Someren E, Rosenberg J (2009) Disturbances in the circadian pattern of activity and sleep after laparoscopic versus open abdominal surgery. Surg Endosc 23:1026–1031. https://doi.org:10.1007/s00464-008-0112-9 Ni P, Dong H, Zhou Q, Wang Y, Sun M, Qian Y, Sun J (2019) Preoperative Sleep Disturbance Exaggerates Surgery-Induced Neuroinflammation and Neuronal Damage in Aged Mice. Mediators Inflamm 2019:8301725. https://doi.org:10.1155/2019/8301725 Ruiz FS, Andersen ML, Guindalini C, Araujo LP, Lopes JD, Tufik S (2017) Sleep influences the immune response and the rejection process alters sleep pattern: Evidence from a skin allograft model in mice. Brain Behav Immun 61:274–288. https://doi.org:10.1016/j.bbi.2016.12.027 Tabassum S, Misrani A, Tabassum S, Ahmed A, Yang L, Long C (2021) Disrupted prefrontal neuronal oscillations and morphology induced by sleep deprivation in young APP/PS1 transgenic AD mice. Brain Res Bull 166:12–20. https://doi.org:10.1016/j.brainresbull.2020.11.003 Althammer F, Grinevich V (2017) Diversity of oxytocin neurons: beyond magno- and parvocellular cell types? J Neuroendocrinol. https://doi.org:10.1111/jne.12549 Swaab DF, Nijveldt F, Pool CW (1975) Distribution of oxytocin and vasopressin in the rat supraoptic and paraventricular nucleus. J Endocrinol 67:461–462. https://doi.org:10.1677/joe.0.0670461 Beurel E, Nemeroff CB (2014) Interaction of stress, corticotropin-releasing factor, arginine vasopressin and behaviour. Curr Top Behav Neurosci 18:67–80. https://doi.org:10.1007/7854_2014_306 Knobloch HS, Charlet A, Hoffmann LC, Eliava M, Khrulev S, Cetin AH, Osten P, Schwarz MK, Seeburg PH, Stoop R, Grinevich V (2012) Evoked axonal oxytocin release in the central amygdala attenuates fear response. Neuron 73:553–566. https://doi.org:10.1016/j.neuron.2011.11.030 Neumann ID, Landgraf R (2012) Balance of brain oxytocin and vasopressin: implications for anxiety, depression, and social behaviors. Trends Neurosci 35:649–659. https://doi.org:10.1016/j.tins.2012.08.004 Braga RI, Panaitescu A, Bădescu S, Zăgrean A-M, Zăgrean L (2014) Intranasal administration of oxytocin alters sleep architecture. Biol Rhythm Res 45:69–75. https://doi.org:10.1080/09291016.2013.797641 Lancel M, Krömer S, Neumann ID (2003) Intracerebral oxytocin modulates sleep-wake behaviour in male rats. Regul Pept 114:145–152. https://doi.org:10.1016/s0167-0115(03)00118-6 Mahoney C, Kroeger D, Grinevich V, Scammell T (2017) 0134 Oxytocin fibers in the lateral hypothalamus promote arousal in a mouse model of PWS. Sleep 40:A50 Venn RM, Bradshaw CJ, Spencer R, Brealey D, Caudwell E, Naughton C, Vedio A, Singer M, Feneck R, Treacher D, Willatts SM, Grounds RM (1999) Preliminary UK experience of dexmedetomidine, a novel agent for postoperative sedation in the intensive care unit. Anaesthesia 54:1136–1142. https://doi.org:10.1046/j.1365-2044.1999.01114.x Nelson LE, Lu J, Guo T, Saper CB, Franks NP, Maze M (2003) The alpha2-adrenoceptor agonist dexmedetomidine converges on an endogenous sleep-promoting pathway to exert its sedative effects. Anesthesiology 98:428–436. https://doi.org:10.1097/00000542-200302000-00024 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 30 Jul, 2024 Read the published version in Neurochemical Research → Version 1 posted Editorial decision: Revision requested 09 Jul, 2024 Reviews received at journal 13 May, 2024 Reviewers agreed at journal 29 Apr, 2024 Reviewers invited by journal 07 Apr, 2024 Editor assigned by journal 04 Apr, 2024 Submission checks completed at journal 02 Apr, 2024 First submitted to journal 02 Apr, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {\"props\":{\"pageProps\":{\"initialData\":{\"identity\":\"rs-4207174\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":false,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":288482983,\"identity\":\"cbf862a2-d313-4e46-993d-0e20b3d96769\",\"order_by\":0,\"name\":\"Ying Zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Ying\",\"middleName\":\"\",\"lastName\":\"Zhang\",\"suffix\":\"\"},{\"id\":288482984,\"identity\":\"29d9a905-085b-4f8d-9927-62efb4ae5bd6\",\"order_by\":1,\"name\":\"Jiaxin Li\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Jiaxin\",\"middleName\":\"\",\"lastName\":\"Li\",\"suffix\":\"\"},{\"id\":288482985,\"identity\":\"631455dd-9241-44ec-9514-c9589f02c26e\",\"order_by\":2,\"name\":\"Wei Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Wei\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":288482986,\"identity\":\"c6b5c5de-3f5d-45d6-b1c1-0dba30bea4fa\",\"order_by\":3,\"name\":\"Daming Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"the Second Affiliated Hospital of Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Daming\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":288482987,\"identity\":\"68806b03-1b63-4b71-a3d6-ae5785c9fc7c\",\"order_by\":4,\"name\":\"Junli Ding\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"First Affiliated Hospital of Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Junli\",\"middleName\":\"\",\"lastName\":\"Ding\",\"suffix\":\"\"},{\"id\":288482988,\"identity\":\"15d886f2-332d-4522-b973-4cc3db9f8e08\",\"order_by\":5,\"name\":\"Liecheng Wang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Anhui Medical University\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Liecheng\",\"middleName\":\"\",\"lastName\":\"Wang\",\"suffix\":\"\"},{\"id\":288482989,\"identity\":\"0ed30b97-31ed-49d8-97ce-1cf947a23419\",\"order_by\":6,\"name\":\"Juan Cheng\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA0UlEQVRIiWNgGAWjYHACNhDBzMbefODAhx+kaOHjOZZ4cGYPCVoY5CRyjA9zsBGhXr6999iDD3/s2NkYcj4cZuBhkOcXO4Bfi8GZc+mGM9uSmdkYzm44XGDBYDhzdgIBLRI5ZtK8DQeY2Rh7NxyewcOQYHCbgBb5GUAtPH+AWph5HhzmYSNCC8MNkBY2oBY2HgbitAD9kiYJ9gsPmwEwkCUI+wUUYhLAEEuWn//48YcPP2zk+aUJOQwYriCQDOVJEFKO0GJHjNJRMApGwSgYoQAA/ENAAaUeYF0AAAAASUVORK5CYII=\",\"orcid\":\"\",\"institution\":\"Anhui Medical University\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Juan\",\"middleName\":\"\",\"lastName\":\"Cheng\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-04-02 13:52:15\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4207174/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4207174/v1\",\"draftVersion\":[],\"editorialEvents\":[{\"content\":\"https://doi.org/10.1007/s11064-024-04221-w\",\"type\":\"published\",\"date\":\"2024-07-30T15:57:08+00:00\"}],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":54269209,\"identity\":\"7f9cf629-372a-46ab-91de-d8ee41700a77\",\"added_by\":\"auto\",\"created_at\":\"2024-04-08 06:13:53\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1208114,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eMicroinjection of DEX into the PVN increased the amount of NREM sleep during ZT14-18 in mice.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003ea\\u003c/strong\\u003e Schematic diagram of the PVN with an implanting guide and internal cannula (left) and implanting EEG/EMG electrode. \\u003cstrong\\u003eb\\u003c/strong\\u003e The flow chart of the EEG/EMG experiment. \\u003cstrong\\u003ec-d\\u003c/strong\\u003e Electroencephalogram power spectrum, electromyogram, and total duration of sleep-wakefulness states of the saline group (\\u003cstrong\\u003ec\\u003c/strong\\u003e) and the DEX group (\\u003cstrong\\u003ed\\u003c/strong\\u003e). \\u003cstrong\\u003ee-g\\u003c/strong\\u003e Percentage of one hour in Wake (\\u003cstrong\\u003ee\\u003c/strong\\u003e), NREM (\\u003cstrong\\u003ef\\u003c/strong\\u003e), and REM (\\u003cstrong\\u003eg\\u003c/strong\\u003e) states in OXT-hM3D(Gq)-mCherry mice and the statistical chart of the duration of three sleep-wakefulness states in ZT14-18. (\\u003cstrong\\u003ee\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA ((F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 3.172， \\u003cem\\u003eP\\u003c/em\\u003e = 0.1729 \\u0026gt; 0.05) and paired Student's t-test (* \\u003cem\\u003eP\\u003c/em\\u003e = 0.0333 \\u0026lt; 0.05; N = 5 mice); \\u003cstrong\\u003ef\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA (F\\u003csub\\u003e(1,4)\\u003c/sub\\u003e = 19.64，* \\u003cem\\u003eP= \\u003c/em\\u003e0.0114 \\u0026lt; 0.05) and paired Student's t-test (* \\u003cem\\u003eP\\u003c/em\\u003e = 0.0316 \\u0026lt; 0.05); N = 4 mice); \\u003cstrong\\u003eg\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA (F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.1362, \\u003cem\\u003eP\\u003c/em\\u003e = 0.7366 \\u0026gt; 0.05) and paired Student's t-test (\\u003cem\\u003eP\\u003c/em\\u003e = 0.5924 \\u0026gt; 0.05); N = 4 mice). \\u003cstrong\\u003eh\\u003c/strong\\u003e EEG PSD of saline (black) and DEX (blue) after saline or DEX injection. a: 4 Hz, b: 8 Hz, c: 13 Hz. (The data were analyzed using two-way repeated-measures ANOVA (F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 5.455, \\u003cem\\u003eP\\u003c/em\\u003e =0.1016 \\u0026gt; 0.05) and Bonferroni’s multiple comparisons test (**** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(1.0-3.3 Hz)\\u003c/sub\\u003e \\u0026lt; 0.0001; *** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.4.Hz)\\u003c/sub\\u003e = 0.002 \\u0026lt; 0.001; *** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.5.Hz)\\u003c/sub\\u003e = 0.001; ** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.6.Hz)\\u003c/sub\\u003e = 0.0011 \\u0026lt; 0.01; ** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.7.Hz)\\u003c/sub\\u003e = 0.0020 \\u0026lt; 0.01; ** \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.8.Hz)\\u003c/sub\\u003e = 0.0024 \\u0026lt; 0.001; * \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(3.9 Hz)\\u003c/sub\\u003e = 0.0102 \\u0026lt; 0.05); N = 5 mice). \\u003cstrong\\u003ei\\u003c/strong\\u003e The mean duration of Wake, NREM and REM in 4 hours. (paired Student's t-test: * \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(Wake)\\u003c/sub\\u003e = 0.0217 \\u0026lt; 0.05;\\u003cem\\u003e \\u003c/em\\u003eWilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(NREM)\\u003c/sub\\u003e = 0.6250 \\u0026gt; 0.05; paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(REM)\\u003c/sub\\u003e = 0.2907 \\u0026gt; 0.05; N = 4 Mice). \\u003cstrong\\u003ej\\u003c/strong\\u003e The transition number of Wake, NREM sleep and REM sleep in 4 hours. (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(Wake to NREM)\\u003c/sub\\u003e = 0.0525 \\u0026gt; 0.05; paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(NREM to Wake)\\u003c/sub\\u003e = 0.0705 \\u0026gt; 0.05; paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(NREM to REM)\\u003c/sub\\u003e = 0.3871 \\u0026gt; 0.05; Wilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(REM to Wake)\\u003c/sub\\u003e \\u0026gt; 0.9999; paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e \\u003csub\\u003e(REM to NREM)\\u003c/sub\\u003e = 0.3396 \\u0026gt; 0.05; N = 4 mice). Data are represented as mean ± SEM.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"figure1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/a0fa61d7be79df8c83980640.png\"},{\"id\":54269210,\"identity\":\"1f684725-fe2e-4c7b-93df-efc389d7169e\",\"added_by\":\"auto\",\"created_at\":\"2024-04-08 06:13:53\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1580039,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eChemogenetic activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons could counteract DEX-induced NREM Sleep.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003ea\\u003c/strong\\u003e Schematic diagram of the PVN with an implanting guide and internal cannulas, and a representation of hM3Dq expressed in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons (scale bar: 200 μm). \\u003cstrong\\u003eb\\u003c/strong\\u003e Current-clamp recording of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons expressing hM3D(Gq) Virus. \\u003cstrong\\u003ec\\u003c/strong\\u003e The flow chart of the EEG/EMG experiment. \\u003cstrong\\u003ed-f\\u003c/strong\\u003e The persentage of Wake(D), NREM(E) and REM(F) of OXT-hM3D(Gq)-mCherry mice and the amount of Wake(D), NREM(E) and REM(F) during ZT2-3. (\\u003cstrong\\u003ed\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA (F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.2.413, \\u003cem\\u003eP\\u003c/em\\u003e = 0.2181 \\u0026gt; 0.05) and paired Student's t-test: (* \\u003cem\\u003eP\\u003c/em\\u003e = 0.0163 \\u0026lt; 0.05); \\u003cstrong\\u003eEe\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA(F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.3620, \\u003cem\\u003eP\\u003c/em\\u003e = 0.5898 \\u0026gt; 0.05) and paired Student’s t-test(*\\u003cem\\u003e P\\u003c/em\\u003e = 0.0291 \\u0026lt; 0.05); \\u003cstrong\\u003ef\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA (F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 5.832, \\u003cem\\u003eP\\u003c/em\\u003e = 0.0946 \\u0026gt; 0.05) and Wilcoxon matched-pairs signed rank test (\\u003cem\\u003eP\\u003c/em\\u003e = 0.1250 \\u0026gt; 0.05); N = 4 mice. \\u003cstrong\\u003eg-h\\u003c/strong\\u003e Electroencephalogram power spectrum, electromyogram, and total duration of sleep-wakefulness states of the saline group (\\u003cstrong\\u003eg\\u003c/strong\\u003e) and CNO group (\\u003cstrong\\u003eh\\u003c/strong\\u003e). \\u003cstrong\\u003ei-k\\u003c/strong\\u003e the percentage of Wake(\\u003cstrong\\u003ei\\u003c/strong\\u003e), NREM(\\u003cstrong\\u003ej\\u003c/strong\\u003e), and REM(\\u003cstrong\\u003ek\\u003c/strong\\u003e) of OXT-hM3D(Gq)-mCherry mice (\\u003cstrong\\u003ei\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA(F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.8715, \\u003cem\\u003eP\\u003c/em\\u003e = 0.4194 \\u0026gt; 0.05) and paired Student’s t-test( * \\u003cem\\u003eP\\u003c/em\\u003e = 0.0333 \\u0026lt; 0.05); \\u003cstrong\\u003ej\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA(F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.8827， \\u003cem\\u003eP\\u003c/em\\u003e = 0.4168 \\u0026gt; 0.05); \\u003cstrong\\u003ek\\u003c/strong\\u003e: The data were analyzed using two-way repeated-measures ANOVA(F\\u003csub\\u003e(1, 3)\\u003c/sub\\u003e = 0.5659,\\u003cem\\u003e P\\u003c/em\\u003e = 0.5066 \\u0026gt; 0.05). \\u003cstrong\\u003el-n\\u003c/strong\\u003e The amount of Wake(\\u003cstrong\\u003el\\u003c/strong\\u003e), NREM(\\u003cstrong\\u003em\\u003c/strong\\u003e), and REM(\\u003cstrong\\u003en\\u003c/strong\\u003e) during ZT15.5, ZT15.5-16.0, 16.0-16.5, 16.0-17.0, 16.0-17.5 (\\u003cstrong\\u003el\\u003c/strong\\u003e: Wilcoxon matched-pairs signed rank test (ZT15.5-16: \\u003cem\\u003eP\\u003c/em\\u003e = 0.5000 \\u0026gt; 0.05) and paired Student’s t-test (ZT16.0-16.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.5924 \\u0026gt; 0.05; ZT16.0-17.0: * \\u003cem\\u003eP\\u003c/em\\u003e = 0.0492\\u0026nbsp; \\u0026lt; 0.05; ZT16.0-17.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.5350 \\u0026gt; 0.05); \\u003cstrong\\u003em\\u003c/strong\\u003e: Wilcoxon matched-pairs signed rank test (ZT15.5-16: \\u003cem\\u003eP\\u003c/em\\u003e = 0.5000 \\u0026gt; 0.05) and paired Student’s t-test (ZT16.0-16.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.3511 \\u0026gt; 0.05; ZT16.0-17.0: * \\u003cem\\u003eP\\u003c/em\\u003e = 0.0329\\u0026nbsp; \\u0026lt; 0.05; ZT16.0-17.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.5250 \\u0026gt; 0.05); \\u003cstrong\\u003en\\u003c/strong\\u003e: paired Student’s t-test (ZT16.0-16.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.6913 \\u0026gt; 0.05; ZT16.0-17.0: \\u003cem\\u003eP\\u003c/em\\u003e = 0.2870 \\u0026gt; 0.05; ZT16.0-17.5: \\u003cem\\u003eP\\u003c/em\\u003e = 0.8750 \\u0026gt; 0.05); N = 4 mice). Data are represented as mean ± SEM.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"figure2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/a56ba584abb0159fe49c33db.png\"},{\"id\":54269208,\"identity\":\"449c711a-93d3-4858-9fc4-30fd15b97b4a\",\"added_by\":\"auto\",\"created_at\":\"2024-04-08 06:13:53\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":667028,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eDEX inhibited the AP frequency of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003ea\\u003c/strong\\u003e Images showing whole-cell recording with a patch pipette in the light transmission view between the bright field and fluorescence of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. \\u003cstrong\\u003eb\\u003c/strong\\u003e Diagram describing electrophysiological characteristics of action potential (horizontal bar: 10 ms; vertical bar: 10 mV). \\u003cstrong\\u003ec\\u003c/strong\\u003e Compared with the aCSF group, AP frequency showed a decreased frequency in the DEX group (Mann Whitney test: * \\u003cem\\u003eP\\u003c/em\\u003e = 0.0279 \\u0026lt; 0.05; N = 5 mice, n = 20-21 cells). \\u003cstrong\\u003ed\\u003c/strong\\u003e No differences were observed in the APD\\u003csub\\u003e50\\u003c/sub\\u003e between the aCSF group and the DEX group. (unpaired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.2687 \\u0026lt; 0.05; N = 5 mice, n = 19-20 cells). \\u003cstrong\\u003ee\\u003c/strong\\u003e ­­No differences were observed in the AP amplitude between the aCSF group and the DEX group. (unpaired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.8554 \\u0026gt; 0.05; N = 5 mice, n = 19-20 cells). \\u003cstrong\\u003ef\\u003c/strong\\u003e No differences were observed in the spike max between the aCSF group and the DEX group. (unpaired Student's t-test: \\u003cem\\u003eP \\u003c/em\\u003e= 0.6308 \\u0026gt; 0.05; N = 5 mice, n = 19-20 cells). \\u003cstrong\\u003eg\\u003c/strong\\u003e No differences were observed in the AHP amplitude between the aCSF group and the DEX group. (Mann Whitney test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.6695 \\u0026gt; 0.05; N = 5 mice, n = 19-20 cells). \\u003cstrong\\u003eh\\u003c/strong\\u003e No differences were observed in the threshold of AP between the aCSF group and the DEX group. (unpaired Student's t-test: \\u003cem\\u003eP \\u003c/em\\u003e= 0.4712 \\u0026gt; 0.05; N = 5 mice, n = 19-20 cells). \\u003cstrong\\u003ei\\u003c/strong\\u003e No differences were observed in the RMP between the aCSF group and the DEX group. (Mann Whitney test: \\u003cem\\u003eP \\u003c/em\\u003e= 0.8508 \\u0026gt; 0.05; N = 5 mice, n = 20-21 Cells). \\u003cstrong\\u003ej\\u003c/strong\\u003e No differences were observed in the threshold of R\\u003csub\\u003ein\\u003c/sub\\u003e between the aCSF group and the DEX group. (unpaired Student's t-test: \\u003cem\\u003eP \\u003c/em\\u003e= 0.1129 \\u0026gt; 0.05; N = 5 mice, n = 18-20 Cells). \\u003cstrong\\u003ek\\u003c/strong\\u003e After -40-0 pA current stimulation, there were no differences in the subtraction of RMP and hyperpolarized membrane potential between the aCSF group and the DEX group. (The data were analyzed using two-way ANOVA (F\\u003csub\\u003e(1, 36)\\u003c/sub\\u003e = 3.360, \\u003cem\\u003eP =\\u003c/em\\u003e 0.0751 \\u0026gt; 0.05; N = 5 mice, n = 18-20 cells). Data are represented as mean ± SEM.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"figure3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/d22604bdb8ca07b4eba3aa8d.png\"},{\"id\":54269212,\"identity\":\"2b7e1237-8da2-41c5-9a09-2489bed425cb\",\"added_by\":\"auto\",\"created_at\":\"2024-04-08 06:13:53\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":323981,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eBlocking the α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors counteracted the DEX-induced inhibiting of AP frequency in PVN\\u003csup\\u003eOXT \\u003c/sup\\u003eneurons.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003ea\\u003c/strong\\u003e The flow chart of the whole-cell patch clamp experiment. \\u003cstrong\\u003eb\\u003c/strong\\u003e No differences were observed in the AP frequency between the RS79948 group and RS79948+DEX group (Wilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.7609 \\u0026gt; 0.05; N = 4 mice, n = 14 cells). \\u003cstrong\\u003ec\\u003c/strong\\u003e No differences were observed in the APD50 between the RS79948 group and RS79948+DEX group (Wilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.4631 \\u0026lt; 0.05; N = 4 mice, n = 14 cells). \\u003cstrong\\u003ed\\u003c/strong\\u003e No differences were observed in the AP amplitude between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.8554 \\u0026gt; 0.05; N = 4 mice, n = 14 cells) \\u003cstrong\\u003eE\\u003c/strong\\u003e No differences were observed in the spike max between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.3009 \\u0026gt; 0.05; N = 4 mice, n = 14 cells). \\u003cstrong\\u003ef\\u003c/strong\\u003e No differences were observed in the AHP amplitude between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.2816 \\u0026gt; 0.05; N = 4 mice, n = 14 cells). \\u003cstrong\\u003eg\\u003c/strong\\u003e No differences were observed in the threshold between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.1600 \\u0026gt; 0.05; N = 4 mice, n = 14 cells). \\u003cstrong\\u003eh\\u003c/strong\\u003e No differences were observed in the RMP between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.6337 \\u0026gt; 0.05; N = 4 mice, n = 12 cells). \\u003cstrong\\u003ei\\u003c/strong\\u003e No differences were observed in the R\\u003csub\\u003ein\\u003c/sub\\u003e between the RS79948 group and RS79948+DEX group (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.8069 \\u0026gt; 0.05; N = 4 mice, n = 12 cells). \\u003cstrong\\u003ej\\u003c/strong\\u003e After -40-0 pA current stimulation, there were no differences in the subtraction of RMP and hyperpolarized membrane potential between the RS79948 group and RS79948+DEX group (The data were analyzed using two-way repeated-measures ANOVA. (F\\u003csub\\u003e(1, 22)\\u003c/sub\\u003e = 0.1479, \\u003cem\\u003eP\\u003c/em\\u003e = 0.1253 \\u0026gt; 0.05; N = 4 mice, n = 12 cells)). Data are represented as mean ± SEM.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"figure4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/c9b3356e3daae7dcef4f96f2.png\"},{\"id\":54269581,\"identity\":\"95b2e418-7d1c-4ffa-b882-8d701101a918\",\"added_by\":\"auto\",\"created_at\":\"2024-04-08 06:21:53\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":410869,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eBlocking the α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors counteracted the DEX-induced decrease in the frequency of sEPSC in PVN\\u003csup\\u003eOXT \\u003c/sup\\u003eneurons.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003ea\\u003c/strong\\u003e The flow chart of the whole-cell patch clamp experiment between the aCSF group and the DEX group. \\u003cstrong\\u003eb\\u003c/strong\\u003e The flow chart of the whole-cell patch clamp experiment between the RS79948 group and the RS79948+DEX group. \\u003cstrong\\u003ec\\u003c/strong\\u003e Cumulative probability plot of the interevent interval of mEPSC (aCSF (black) and with DEX (blue)). Compared to the aCSF group, the frequency declined in the DEX group (Wilcoxon matched-pairs signed rank test: * \\u003cem\\u003eP\\u003c/em\\u003e = 0.0121 \\u0026lt; 0.05; N = 4 mice, n = 12 cells). \\u003cstrong\\u003ed\\u003c/strong\\u003e There were no differences in the normalized probability and the mean of the amplitude of sEPSC between the aCSF group and the DEX group. (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.6248 \\u0026gt; 0.05; N = 4 mice, n = 12 cells). \\u003cstrong\\u003ee\\u003c/strong\\u003e Cumulative probability plot of the interevent interval of sIPSC (aCSF (black) and with DEX (blue)). There were no differences in the frequency of sIPSC between the aCSF group and the DEX group. (Wilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.4246 \\u0026gt; 0.05; N = 4 mice, n = 13 cells). \\u003cstrong\\u003ef\\u003c/strong\\u003e There were no differences in the normalized probability and the mean of the amplitude of sIPSC between the aCSF group and the DEX group. (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.8086 \\u0026gt; 0.05; N = 4 mice, n = 13 cells). \\u003cstrong\\u003eg\\u003c/strong\\u003e Cumulative probability plot of the interevent interval of sEPSC (RS79948 (black) and with RS79948+DEX (red)). There were no differences in the frequency of sEPSC between the RS79948 group and the RS79948+DEX group. (Wilcoxon matched-pairs signed rank test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.4143 \\u0026gt; 0.05; N = 4 mice, n = 13 cells). \\u003cstrong\\u003eh\\u003c/strong\\u003e There were no differences in the normalized probability and the mean of the amplitude of sEPSC between the RS79948 group and the RS79948+DEX group. (paired Student's t-test: \\u003cem\\u003eP\\u003c/em\\u003e = 0.4930 \\u0026gt; 0.05; N = 4 mice, n = 13 cells). Data are represented as mean ± SEM.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"figure5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/1e64086a5a979eb7b4d945e8.png\"},{\"id\":61793406,\"identity\":\"3d048c33-030f-483d-af48-dc470f274a38\",\"added_by\":\"auto\",\"created_at\":\"2024-08-05 16:12:09\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":4603403,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4207174/v1/d57bbad4-8a56-4816-88ea-03f86154ce84.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"Dexmedetomidine promotes NREM sleep by depressing the neuronal activity of OXT neurons in the PVN\",\"fulltext\":[{\"header\":\"1 Introduction\",\"content\":\"\\u003cp\\u003eDexmedetomidine (DEX) is a highly specific α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoreceptor agonist, with a 1620 times affinity of binding to α\\u003csub\\u003e1\\u003c/sub\\u003e adrenoreceptors [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors are a class of heterotrimeric guanine nucleotide-binding protein receptors (GPCR) [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e], which play a wide role in regulating physiological functions, including heart rate, blood pressure, blood glucose, insulin homeostasis, and analgesia. They are involved in presynaptic feedback inhibition mediating the release of neurotransmitters from the terminal of norepinephrine synapses [\\u003cspan additionalcitationids=\\\"CR4\\\" citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR5\\\" class=\\\"CitationRef\\\"\\u003e5\\u003c/span\\u003e]. As an anesthetic drug, DEX has the characteristics of sedation and analgesia with few side effects and a wide application range[\\u003cspan additionalcitationids=\\\"CR7 CR8 CR9 CR10 CR11\\\" citationid=\\\"CR6\\\" class=\\\"CitationRef\\\"\\u003e6\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e]. The specific role of DEX in postoperative sleep and its related mechanisms are poorly understood.\\u003c/p\\u003e \\u003cp\\u003eAs a sedative-hypnotic drug, DEX has a certain impact on the sleep-wake \\u0026ldquo;switches\\u0026rdquo; [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. Intraperitoneal injection of DEX can increase the c-Fos expression in the overlapping areas of the preoptic area (POA) and diaphragm of the hypothalamus [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e], ventral tegmental area (VTA) [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e], and area postrema (AP*) [\\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e] and decrease the c-Fos expression in the supraoptic nucleus (SON) [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e], primary motor cortex (M1), and secondary motor cortex (M2) [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e], and these brain regions are associated with the modulation of sleep-wakefulness [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e]. Studies have shown that DEX can increase the calcium signal intensity of dopamine (DA) neurons in the VTA (VTA\\u003csup\\u003eDA\\u003c/sup\\u003e). DEX can increase the excitability of VTA\\u003csup\\u003eDA\\u003c/sup\\u003e neurons through α adrenoreceptors. It also increases the concentration of DA neurotransmitters in the medial prefrontal cortex (MPFC) and nucleus accumbens (NAc), an effect that may be related to the sedative arousal properties of DEX [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]. The activation of VTA\\u003csup\\u003eDA\\u003c/sup\\u003e neurons can induce the state change from an unconscious anesthetic to wakefulness, triggering a strong awakening-promoting effect [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e], which may be the mechanism of DEX-induced rapid awakening after sedation.\\u003c/p\\u003e \\u003cp\\u003eRecently, it was found that vesicular glutamate transporter 2 (Vglut2), oxytocin (OXT), prodynorphin (PDYN), corticotropin-releasing hormone (CRH), and vasopressin (AVP) neurons in the paraventricular nucleus of the hypothalamus (PVN) could induce different levels of arousal through chemogenetic activation [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. PVN is composed of three types of neurons: the large cells (Magno), the small cells (Parvocellular), and the long projection neurons, which express OXT, PDYN, CRH, et.al. Almost one-third of neurons in PVN are expressing OXT (PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e) mainly distributed in the middle part of PVN. [\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e]. AVP and OXT neurons are involved in the regulation of physiology and behavior, including stress response and social behavior. Sleep-wakefulness has an essential attribution to these adaptive behaviors. Our previous study showed that DEX could depress the c-Fos expression of AVP neurons in PVN and reduce the concentration of AVP in serum [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. It has been found that photogenetic and chemogenetic activation of PVN\\u003csup\\u003eAVP\\u003c/sup\\u003e neurons significantly increases the amount of wake in mice [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e]. PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons are an important component in the regulation of sleep and awakening [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. Studies have shown that chemogenetic activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons could increase the amount of wake and decrease the amount of NREM sleep and rapid eye movement (REM) sleep [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. Here, we aim to investigate the roles of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons in the regulation of sleep homeostasis by DEX. We used the chemogenetic methods to activate PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons after canula administration of DEX to PVN, we found that activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons could counteract the increase of NREM sleep time induced by DEX. In addition, we found that DEX inhibited the frequency of action potential (AP) and spontaneous excitatory postsynaptic current (sEPSC) of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors. It is suggested that DEX might maintain sleep homeostasis by inhibiting excitatory synaptic signaling in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons.\\u003c/p\\u003e\"},{\"header\":\"2 Materials and Methods\",\"content\":\" \\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.1 Mice\\u003c/h2\\u003e \\u003cp\\u003e The animal experimental procedures underwent rigorous scrutiny and approval by the Laboratory Animal Management Office of Laboratory Animals at Anhui Medical University, adhering strictly to the pertinent ethical directives established by the Institutional Animal Care Unit Committee of Anhui Medical University, under project identifier LLSC20190763. OXT-IRES-Cre mice (stock No. 024234; Jackson Laboratory, Bar Harbor, ME) and Ai9 mice (stock No. 007909; Jackson Laboratory, Bar Harbor, ME) were generously provided by Professor Tian Xue from the University of Science and Technology of China. Housing conditions for all mice were maintained under a 12-hour light-dark cycle (lights on at 08:00, illuminance ranging from 150\\u0026ndash;200 lux), with ad libitum access to food and water, and a room temperature (RT) set at 25\\u0026deg;C.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec4\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.2 Surgery and virus injection\\u003c/h2\\u003e \\u003cp\\u003eMice were deeply anesthetized with 50 mg/kg sodium pentobarbital and positioned in a stereotaxic frame (RWD Life Science, Shenzhen, China) with sterilized instruments. Utilizing coordinates from the mouse brain atlas[\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e], a microinjection needle delivered 200 nL of rAAV-EF1α-DIO-hM3D(Gq)-mCherry-WPREs (\\u0026ge;\\u0026thinsp;5.00E\\u0026thinsp;+\\u0026thinsp;12 viral particles/mL, Brainvta, Wuhan, China) bilaterally into the PVN at a rate of 40 nL/min (M/L: \\u0026plusmn; 0.30 mm, A/P: -0.6 mm, D/V: -4.6). The guide cannula (RWD Life Science, Shenzhen, China) was implanted in the PVN with sagittal plane tilting 5\\u0026deg;(M/L: 1.0 mm, A/P: -0.6 mm, D/V: -4.6) and secured with light-curable resin (BJBCJB, Sino-Dentex, Jilin, China). The length of internal cannulas (RWD Life Science, Shenzhen, China), is equal to guide cannulas. EEG electrodes were inserted into the skull (+\\u0026thinsp;1.5 mm Bregma, -1.5 mm midline \\u0026ndash; first recording electrode; +1.5 mm Lambda, +\\u0026thinsp;1.5 mm midline \\u0026ndash; second recording electrode), while EMG electrodes were placed in neck musculature. All electrodes were affixed with light-cured resin and quick self-curing acrylic resin (SC01, Yamahahi Dental, Japan). Surgical sites were sutured and disinfected. Post-surgery, mice were housed individually.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec5\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.3 EEG/EMG recordings and analysis\\u003c/h2\\u003e \\u003cp\\u003eThe animals were housed individually in an EEG/EMG recording cage under a 12-hour light:12-hour dark cycle (with light onset at 08:00 and offset at 20:00) for 3 days of acclimation. Electrical signals were recorded at 1 kHz, amplified with the MP150 system (BIOPAC, California, USA), and digitized using Acknowledge 4.2 software (BIOPAC, California, USA). Automated analysis categorized EEG/EMG signals into Wake, NREM sleep, and REM sleep states every 4 seconds using the Lunion Stage AI engine (LunionData, Shanghai, China), with manual verification and correction. EEG signals were filtered between 0.3\\u0026ndash;35 Hz and EMG signals between 10\\u0026ndash;200 Hz.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec6\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.4 Electrophysiology\\u003c/h2\\u003e \\u003cdiv id=\\\"Sec7\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.4.1 Slice preparation\\u003c/h2\\u003e \\u003cp\\u003eOXT::Ai9 mice (8\\u0026ndash;12 weeks old) were anesthetized with 1% isoflurane (RWD Life Science, Shenzhen, China) and decapitated. The brain was swiftly excised and immersed in a freezing oxygenated (95% O\\u003csub\\u003e2\\u003c/sub\\u003e and 5% CO\\u003csub\\u003e2\\u003c/sub\\u003e) cutting solution containing N-methyl-D-glucamine, 1-deoxy-1-(methylamine)-D-sorbito, glucamine (NMDG) cutting solution (in mM: 92 NMDG, 1.2 potassium chloride (KCl), 1.2 potassium dihydrogen phosphate (KH\\u003csub\\u003e2\\u003c/sub\\u003ePO\\u003csub\\u003e4\\u003c/sub\\u003e), 30 sodium bicarbonate (NaHCO\\u003csub\\u003e3\\u003c/sub\\u003e), 25 D-glucose, 20 4-hydroxyethyl piperazine ethanesulfonic acid (HEPES), 5 L-ascorbic acid, 3 sodium pyruvate, 2 thiourea, 10 magnesium sulfate (MgSO\\u003csub\\u003e4\\u003c/sub\\u003e) and 0.5 calcium chloride (CaCl\\u003csub\\u003e2\\u003c/sub\\u003e); the pH adjusted to 7.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.1 with hydrogen chloride (HCl), the osmolarity adjusted to 310\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5 mOsmol/L with NMDG). Horizontal brain slices (300 \\u0026micro;m) were then obtained using a vibrating microtome (VT1200S, Leica, Germany). These slices were transferred to oxygenated (95% O\\u003csub\\u003e2\\u003c/sub\\u003e and 5% CO\\u003csub\\u003e2\\u003c/sub\\u003e) aCSF (mM: 125 NaCl, 1.25 KCl, 2 CaCl\\u003csub\\u003e2\\u003c/sub\\u003e, 1 MgCl\\u003csub\\u003e2\\u003c/sub\\u003e, 1.25 KH\\u003csub\\u003e2\\u003c/sub\\u003ePO\\u003csub\\u003e4\\u003c/sub\\u003e, 25 NaHCO\\u003csub\\u003e3\\u003c/sub\\u003e, 0.4 Na-pyruvate, 0.08 L-ascorbic acid, 25 D-glucose), and allowed incubating for 40\\u0026ndash;45 minutes at 36.6 ℃ before recording.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec8\\\" class=\\\"Section3\\\"\\u003e \\u003ch2\\u003e2.4.2 Whole-cell recordings\\u003c/h2\\u003e \\u003cp\\u003eWhole-cell recordings were performed at RT in oxygenated (95% O\\u003csub\\u003e2\\u003c/sub\\u003e and 5% CO\\u003csub\\u003e2\\u003c/sub\\u003e) aCSF solution with constant perfusion at a rate of 1\\u0026ndash;2 mL/min. For action potential (AP) recording, the electrodes were filled with a potassium-based internal solution (in mM: 125 K-gluconate, 10 KCl, 10 HEPES, 0.1EGTA, 5 ATP-Mg, 0.5GTP-Na; the pH adjusted to 7.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.1 with potassium hydroxide (KOH); the osmolarity adjusted to 310\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5 mOsmol/kg with K-gluconate). For sEPSCs recording, the electrodes were filled with a CsMethane sulfonate-based internal solution containing (in mM: 135 CsMethane sulfonate, 10 cesium chloride (CsCl), 10 HEPES, 1EGTA, 5 ATP-Mg, 0.5 GTP-Na, the pH was adjusted to 7.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.1 using cesium hydroxide (CsOH), and the osmolarity was adjusted to 310\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5 mOsmol/L using CsMethane sulfonate). The aCSF with 50 \\u0026micro;M picrotoxin (PTX, HY-101391, MedChemExpress, USA) was recorded sEPSCs. For sIPSCs recording, the electrodes were filled with a cesium-based internal solution containing (in mM: 140 CsCl, 10 HEPES, 1EGTA, 5 ATP-Mg, 0.5 GTP-Na; the pH was adjusted to 7.2\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;0.1 using CsOH, and the osmolarity was adjusted to 310\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;5 mOsmol/L using CsCl). The aCSF with 10 \\u0026micro;M 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, ab120017, Abcam, UK) and 20 \\u0026micro;M 5-phospho-D-norvaline (D-AP5, ab120003, Abcam, UK) were recorded sIPSCs. sEPSCs were recorded in the presence of the GABA\\u003csub\\u003eA\\u003c/sub\\u003e receptor antagonist Picrotoxin (PTX) and sIPSCs were recorded in the presence of AMPA and NMDA receptor antagonists CNQX and D-AP5 in the aCSF. The neurons were held at -70 mV. The resistance of the patch pipette was 5\\u0026ndash;7 MΩ. A 40\\u0026times; water microscope (TH4-200 and U-RFL-T microscope, Olympus, Japan) was used to visualize the cells. Data were collected by using an Axopatch 700B patch clamp amplifier, Digidatal 1550B data acquisition system, and pClamp10.7 software (Molecular Devices, USA). Data were sampled at 20 kHz, and low-pass filtered with a cutoff frequency of 1 kHz in the current recording. Clampfit 10.7 (Molecular Devices, USA) was used to analyze the electrophysiological properties of the cells. If the hold current of the patch neuron exceeds 30 pA during whole-cell recording or the change of Access resistance (Ra) exceeds 20% during recording, statistics will not be included in the statistical analysis. Igor Pro (WaveMetrics, USA) was used to make statistical charts and representative charts.\\u003c/p\\u003e \\u003c/div\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec9\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.5 Drug administration\\u003c/h2\\u003e \\u003cp\\u003eIn the EEG/EMG recording, OXT-hM3D(Gq)-mCherry mice were recorded from Zeitgeber time (ZT) 14 to ZT0, after the cannula administration of saline or 2 \\u0026micro;M DEX to investigate the DEX-induced effect of sleep-wakefulness states (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eb). OXT-hM3D(Gq)-mCherry mice were injected 1\\u0026micro;L 2 \\u0026micro;M DEX into internal cannulas at ZT14(22:00) and i.p injected saline or CNO at ZT15.5(23:30) to investigate the effect of sleep-wakefulness states in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ec) [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003eElectrophysiological characteristics of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons of APs were captured using the whole-cell patch-clamp method in brain slices immersed in the artificial cerebrospinal fluid (aCSF) with or without 2 \\u0026micro;M DEX. RS79948, an α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenergic receptor antagonist, was utilized to block adrenergic signaling [\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e]. Following a 2-minute stabilization, PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons were microinjected with 1 mM RS79948 for 30 seconds, followed by a 3-minute recording period. Subsequently, neurons received a combined microinjection of 1 mM RS79948 and 2 \\u0026micro;M DEX for 30 seconds, with a further 3-minute recording. (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003ea). To assess the impact of DEX on neuronal signal transmission, sEPSCs and spontaneous inhibitory postsynaptic currents (sIPSCs) were recorded. Initially, neurons were stabilized for 2 minutes, then microinjected with aCSF for 30 seconds, and data were recorded for 2 minutes. This was followed by 30-second microinjection of 2 \\u0026micro;M DEX, with a 2-minute recording period (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003ea). To determine the factors of the observed sEPSCs reduction, neurons were stabilized for 2 minutes, microinjected with 1 mM RS79948 for 30 seconds, and recorded for 1 minute, followed by a co-injection of 1 mM RS79948 and 2 \\u0026micro;M DEX for 30 seconds, with a final 1-minute recording period (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003eb).\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e2.6 Statistical Analysis\\u003c/h2\\u003e \\u003cp\\u003eData are presented as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;standard error of the mean (SEM). The normality of data was assessed using Shapiro-Wilk analysis prior to t-tests. Statistical comparisons between two groups were conducted using paired or unpaired Student's t-tests, Wilcoxon matched-pairs signed rank test, or Mann-Whitney test as appropriate. Multiple parameter comparisons were performed using two-way repeated-measures ANOVA, followed by Bonferroni's multiple comparison test. Prism 10.0 software (GraphPad Software, San Diego, CA, USA) was utilized for all statistical analyses. \\u003cem\\u003eP\\u003c/em\\u003e\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05 was considered statistically significant.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"3 Result\",\"content\":\"\\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.1 DEX increased NREM sleep time and decreased the average duration of wakefulness.\\u003c/h2\\u003e \\u003cp\\u003eFor further exploration of the mechanism of DEX in the sleep-wakefulness cycle, EEG/EMG was divided into three states: NREM sleep, REM sleep, and wake. A guide cannula for drug administration was implanted in the PVN. The mice were cannula injected 1\\u0026micro;L saline or 2 \\u0026micro;M DEX into PVN with a speed of 1\\u0026micro;L/min (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ea). We used the EEG/EMG recording to analyze the sleep-wakefulness phases in mice between the saline group and the DEX group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ec-d). The results showed that the amount of wake during ZT14-18 was decreased (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ee), the amount of NREM sleep during ZT14-18 was increased (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ef), and the amount of REM sleep during ZT14-18 had no differences (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eg) in the DEX group, compared with the saline group., EEG Power spectrum density (PSD) in the DEX group decreased significantly from 1.0-3.9Hz within 4 hours, compared with the saline group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eh). DEX resulted in a decrease in the mean duration of wake (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ei), but the number of sleep transitions had no differences (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003ej).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.2 Chemogenetic activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons counteracted DEX-induced NREM sleep and increased arousal.\\u003c/h2\\u003e \\u003cp\\u003eWe injected rAAV-EF1α-DIO-hM3D(Gq)-mCherry-WPREs into PVN to infect OXT neurons (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ea). Patch clamp recording was used to certify the effectiveness of the virus. The OXT-hM3D(Gq)-expressing neurons were puff with the aCSF and 5\\u0026micro;m clozapine-N-oxide (CNO, Brainvta, Wuhan, China) for 5 minutes respectively, indicating that neurons could be successfully activated by CNO (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eb). The results of i.p. injection of saline or 2 mg/kg CNO into ZT2 showed that activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons in the CNO group could increase the amount of wake and decrease NREM sleep in 1 hour, while REM sleep time had no differences (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ed-f). EEG/EMG was recorded for 10 hours from ZT14 (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003eg, h). After 30 minutes of CNO injection, the time of three sleep states of 0.5, 1, and 1.5 hours were counted. Compared with the saline group, after chemogenetic inhibition of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons, the amount of wake during ZT16-17 in the CNO group was increased, while the amount of NREM time was decreased and the amount of REM had no differences (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003ei-m). In conclusion, the DEX-induced increase in NREM sleep was counteracted by the activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons.\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec14\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.3 DEX inhibited the AP frequency of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons.\\u003c/h2\\u003e \\u003cp\\u003eTo further investigate the mechanism of the effect of DEX on PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons on sleep homeostasis, we prepared brain slices containing PVN from OXT::Ai9 mice during ZT13 and ZT14. During electrophysiological recording, fluorescent images of recorded PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons were collected (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ea). We examined the firing properties of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons before and after drug administration, such as the AP duration were measured at 50% repolarization (APD\\u003csub\\u003e50\\u003c/sub\\u003e), AP amplitude, spike max, afterhyperpolarization (AHP) amplitudes, thresholds, resting cell membrane potential (RMP), membrane input resistance (R\\u003csub\\u003ein\\u003c/sub\\u003e) (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003eb). The results showed that the AP frequency of OXT neurons in the DEX group was significantly reduced, compared to the aCSF group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ec). However, there were no significant differences in APD\\u003csub\\u003e50\\u003c/sub\\u003e, AP amplitude, spike max, AHP, threshold, RMP, and R\\u003csub\\u003ein\\u003c/sub\\u003e between the aCSF group and the DEX group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ed-j). When the neurons received stimulation from \\u0026minus;\\u0026thinsp;40 pA to 0 pA, the DEX group showed a tendency to decrease the potential difference, compared with the aCSF group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig5\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003ek). DEX might reduce the rate of ion influx. Further analysis of the changes in membrane potential and resistance will help to reveal the finer regulatory mechanism of DEX on the function of OXT neurons.\\u003c/p\\u003e \\u003cp\\u003e \\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec15\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003e3.4 DEX decreased the AP and sEPSC frequencies of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors.\\u003c/h2\\u003e \\u003cp\\u003eAs mentioned above, DEX is a highly potent α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors agonist. We recorded the effect of blocking the receptor-mediated signaling pathway on the electrophysiological characteristics of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons by patch-clamp experiments. The results showed that the AP frequency, APD\\u003csub\\u003e50\\u003c/sub\\u003e, AP amplitude, spike max, AHP amplitude, threshold, RMP, and R\\u003csub\\u003ein\\u003c/sub\\u003e have not changed in the RS79948\\u0026thinsp;+\\u0026thinsp;DEX group, compared with the RS79948 group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig3\\\" class=\\\"InternalRef\\\"\\u003e4\\u003c/span\\u003eb-j). This suggests that DEX might inhibited the AP frequency in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors. Subsequently, sEPSCs and sIPSCs were recorded to explore whether DEX could influence the neuronal signal transmission of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. We observed a significant decrease in the frequency of sEPSC in the PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons between the aCSF group and the DEX group (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003ec), and no significant difference between the mean amplitude of sEPSC (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003ed). DEX did not show significant effects on both the frequency and amplitude of sIPSC (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003ee, f). Last, we puffed RS79948 to both the aCSF group and the DEX group and recorded sEPSCs and sIPSCs in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. We found that RS79948 blocked the decreased frequency of sEPSC induced by DEX, with no difference in the amplitude of sEPSC (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig4\\\" class=\\\"InternalRef\\\"\\u003e5\\u003c/span\\u003eg, h). Combined with the above results, our study shows that DEX has a significant effect on modulating AP and excitatory synaptic transmission in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"4 Discussion\",\"content\":\"\\u003cp\\u003eClinical studies show that severe sleep disorders and delirium often occur after surgery [\\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e]. Patients exhibit sleep deprivation, sleep fragmentation, and a reduction in the duration of slow-wave sleep and REM sleep [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e]. The most significant features of postoperative delirium include cognitive deficits, dysfunction of circadian rhythm, and emotional disorders. Studies have shown that the DEX has characteristics of low side effects, which could improve postoperative sleep quality, and reduce the incidence of postoperative delirium after surgery [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e]. Disturbance of sleep structure is a serious problem for postoperative patients, which may lead to prolonged postoperative recovery time [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e] and increase the incidence of cardiovascular diseases, cognitive dysfunctions [\\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e], and impaired immune functions [\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e]. Therefore, we hope to find out the mechanism of DEX on the modulation of sleep homeostasis, further reducing the incidence of delirium.\\u003c/p\\u003e \\u003cp\\u003eStudies showed that oral administration of DEX can increase the duration of NREM sleep within 6 hours, and decrease the duration of REM sleep within 6 hours[\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e]. In a sleep deprivation experiment, the polysomnography results show that the delta wave spectrum in NREM sleep would increase during the recovery period after sleep deprivation [\\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e]. DEX-induced sedation is similar to deep restorative sleep after sleep deprivation. Unlike the classical hypnotic drug diazepam, oral administration of DEX could increase the power of the delta spectrum in NREM sleep EEG, especially at high-frequency delta waves of 1.75\\u0026ndash;3.25 Hz, while the power of low-frequency delta waves of 0.5-1.0 Hz was reduced. The same phenomenon appeared in our study, the cannula injection of DEX to PVN could increase the amount of NREM sleep time and decrease the mean duration of wakefulness, and DEX decreased the delta spectrum at 1.0-3.9Hz. If subsequent studies can find the effect of DEX on different delta wave frequencies, it will be helpful to further analyze the regulatory effect of DEX on NREM sleep [\\u003cspan citationid=\\\"CR13\\\" class=\\\"CitationRef\\\"\\u003e13\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eOXT-expressing neurons are predominantly located in the PVN and supraoptic nucleus (SON) of the mammalian hypothalamus. OXT plays a crucial role in modulates responses to stress, fear, and social behaviors[\\u003cspan additionalcitationids=\\\"CR39 CR40 CR41\\\" citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e]. Little research has investigated the influence of OXT on sleep-wakefulness. Interestingly, OXT might have a dual mechanism of action in dependence of the physiological state on sleep regulation. Long-term intranasal administration of OXT reduced sleep latencies and increased the sleep efficiency and percentage of REM sleep episodes in human [\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e]. A study in mice showed that acute lateral ventricle (ICV) infusion of OXT delayed sleep onset latency, which resulted in a transient reduction of NREM sleep and REM sleep, and augmented high-frequency activity in NREM sleep [\\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e44\\u003c/span\\u003e]. Optogenetic activation of OXT neurons in PVN can promote arousal [\\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e45\\u003c/span\\u003e]. The same phenomenon was found in our study, the chemogenetic activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons could increase the amount of wake. We found that the activation of wakeful-promoting PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through the chemogenetic method could decrease the one-hour amount of NREM sleep induced by DEX. DEX-induced sedation has the property of deep restorative sleep after sleep deprivation [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e] and is easy to wake up [\\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e46\\u003c/span\\u003e]. Whether this process was modulated by working on OXT neurons needs further studies.\\u003c/p\\u003e \\u003cp\\u003eDifferent firing patterns of neurons dictate the amount and timing of neuropeptide release, and the activity of the network affects the neurons' integrative properties. The previous study indicates DEX can increase or decrease delta and theta wave percentage through chemogenetic inhibition or activation of VTA\\u003csup\\u003eDA\\u003c/sup\\u003e neurons. DEX increases the firing rate of VTA\\u003csup\\u003eDA\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e adrenoceptors [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e]. DEX is widely recognized for its sedative properties and is predominantly believed to exert its effects by engaging the norepinephrine system. Extensive research indicates that DEX interacts with α\\u003csub\\u003e2\\u003c/sub\\u003e adrenoceptors located on the norepinephrine-producing neurons of the LC. This interaction triggers a hyperpolarization of the neuronal cell membranes via a Gi protein-mediated pathway. As a result, the release of the stimulating neurotransmitter norepinephrine is diminished across the brain, culminating in a state of sedation[\\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e47\\u003c/span\\u003e]. Here, we examined the influence of DEX on the membrane characteristics and synaptic properties of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. We found DEX decreases the frequency of AP in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. The chemogenetic activation of PVN\\u003csup\\u003eVglut2\\u003c/sup\\u003e neurons can increase arousal indicate glutamate signaling pathway participates in modulating the sleep-wakefulness cycle [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e], and in our study, we also found DEX could decrease the frequency of sEPSC of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. Therefore, DEX might reduce the rate of ion influx by bounding α\\u003csub\\u003e2\\u003c/sub\\u003e adrenoceptors, resulting in decreased excitability of the PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons.\\u003c/p\\u003e \\u003cp\\u003eIn conclusion, these findings highlighted DEX increased NREM sleep by inhibiting PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons, which promoted arousal and decreased the frequency of AP and sEPSC of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons. The results reveal the potential role of modulation of DEX in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons and provide new ideas for the treatment of postoperative delirium prevention. However, further research is still needed to resolve the molecular mechanisms of DEX and its more far-reaching effects on neuronal networks.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003ch2\\u003eAcknowledgments\\u003c/h2\\u003e\\n\\u003cp\\u003eThe authors thank Dandan Zang, Cheng Qian, and Guanjun Chen of the Center for Scientific Research of Anhui Medical University for their valuable help in our experiments.\\u003c/p\\u003e\\n\\u003ch2\\u003eAuthor Contributions\\u003c/h2\\u003e\\n\\u003cp\\u003eJuan Cheng and Liecheng Wang supervised and designed the work; Ying Zhang and Juan Cheng wrote and revised the manuscript; Ying Zhang and Jiaxin Li completed the patch-clamp experiment, EEG/EMG recording and gene identification. Ying Zhang, Jiaxin Li, Wei Wang, Daming Wang, and Junli Ding analyzed the experiment data and bred the mice.\\u003c/p\\u003e\\n\\u003ch2\\u003eData Availability Statement:\\u003c/h2\\u003e\\n\\u003cp\\u003eRaw data will be made available on request to the corresponding author.\\u003c/p\\u003e\\n\\u003ch2\\u003eDeclaration of Conflicting Interests\\u003c/h2\\u003e\\n\\u003cp\\u003eThe author(s) declared no potential conflicts of interest concerning the research, authorship, and/or publication of this article.\\u003c/p\\u003e\\n\\u003ch2\\u003eFunding\\u003c/h2\\u003e\\n\\u003cp\\u003eThe author(s) disclosed receipt of the following financial support for the research, authorship, and/or publication of this article: Our work was supported by the National Natural Science Foundation of China (31800997 to Juan Cheng, 81971236, 81571293 to Liecheng Wang), the Natural Science Foundation of Universities of Anhui Province (2022AH050783 to Juan Cheng), Anhui Provincial Natural Science Foundation (2308085MC83 to Juan Cheng), Research Fund of Anhui Institute of translational medicine (2023zhyx-C71 to Daming Wang) , Basic and Clinical Cooperative Research Promotion Program of Anhui Medical University (2020xkjT027 to Junli Ding).\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eWeerink MAS, Struys M, Hannivoort LN, Barends CRM, Absalom AR, Colin P (2017) Clinical Pharmacokinetics and Pharmacodynamics of Dexmedetomidine. Clin Pharmacokinet 56:893\\u0026ndash;913. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1007/s40262-017-0507-7\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1007/s40262-017-0507-7\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLink R, Daunt D, Barsh G, Chruscinski A, Kobilka B (1992) Cloning of two mouse genes encoding alpha 2-adrenergic receptor subtypes and identification of a single amino acid in the mouse alpha 2-C10 homolog responsible for an interspecies variation in antagonist binding. Mol Pharmacol 42:16\\u0026ndash;27\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWang CD, Buck MA, Fraser CM (1991) Site-directed mutagenesis of alpha 2A-adrenergic receptors: identification of amino acids involved in ligand binding and receptor activation by agonists. Mol Pharmacol 40:168\\u0026ndash;179\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eRamos BP, Arnsten AF (2007) Adrenergic pharmacology and cognition: focus on the prefrontal cortex. Pharmacol Ther 113:523\\u0026ndash;536\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMaletic V, Eramo A, Gwin K, Offord SJ, Duffy RA (2017) The Role of Norepinephrine and Its α-Adrenergic Receptors in the Pathophysiology and Treatment of Major Depressive Disorder and Schizophrenia: A Systematic Review. Front Psychiatry 8:42. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.3389/fpsyt.2017.00042\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.3389/fpsyt.2017.00042\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSun S, Wang J, Bao N, Chen Y, Wang J (2017) Comparison of dexmedetomidine and fentanyl as local anesthetic adjuvants in spinal anesthesia: a systematic review and meta-analysis of randomized controlled trials. Drug Des Devel Ther 11:3413\\u0026ndash;3424. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.2147/DDDT.S146092\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.2147/DDDT.S146092\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBarends CR, Absalom A, van Minnen B, Vissink A, Visser A (2017) Dexmedetomidine versus Midazolam in Procedural Sedation. A Systematic Review of Efficacy and Safety. PLoS ONE 12:e0169525. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1371/journal.pone.0169525\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1371/journal.pone.0169525\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLin Y, Zhang R, Shen W, Chen Q, Zhu Y, Li J, Chi W, Gan X (2020) Dexmedetomidine versus other sedatives for non-painful pediatric examinations: A systematic review and meta-analysis of randomized controlled trials. J Clin Anesth 62:109736. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.jclinane.2020.109736\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.jclinane.2020.109736\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eV ISSRG, W JK, VA. D MM (1988) Dexmedetomidine Diminishes Halothane Anesthetic Requirements in Rats Through a PostsynapticAlphaz Adrenergic Receptor. Anesthesiology\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBrown EN, Pavone KJ, Naranjo M (2018) Multimodal General Anesthesia: Theory and Practice. Anesth Analg 127:1246\\u0026ndash;1258. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1213/ane.0000000000003668\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1213/ane.0000000000003668\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHall JE, Uhrich TD, Barney JA, Arain SR, Ebert TJ (2000) Sedative, amnestic, and analgesic properties of small-dose dexmedetomidine infusions. Anesth Analg 90:699\\u0026ndash;705. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1097/00000539-200003000-00035\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1097/00000539-200003000-00035\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLobo FA, Wagemakers M, Absalom AR (2016) Anaesthesia for awake craniotomy. Br J Anaesth 116:740\\u0026ndash;744. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1093/bja/aew113\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1093/bja/aew113\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eFeng ZX, Dong H, Qu WM, Zhang W (2018) Oral Delivered Dexmedetomidine Promotes and Consolidates Non-rapid Eye Movement Sleep via Sleep-Wake Regulation Systems in Mice. Front Pharmacol 9:1196. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.3389/fphar.2018.01196\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.3389/fphar.2018.01196\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhang Z, Ferretti V, Guntan I, Moro A, Steinberg EA, Ye Z, Zecharia AY, Yu X, Vyssotski AL, Brickley SG, Yustos R, Pillidge ZE, Harding EC, Wisden W, Franks NP (2015) Neuronal ensembles sufficient for recovery sleep and the sedative actions of alpha2 adrenergic agonists. Nat Neurosci 18:553\\u0026ndash;561. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1038/nn.3957\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1038/nn.3957\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eQiu G, Wu Y, Yang Z, Li L, Zhu X, Wang Y, Sun W, Dong H, Li Y, Hu J (2020) Dexmedetomidine Activation of Dopamine Neurons in the Ventral Tegmental Area Attenuates the Depth of Sedation in Mice. Anesthesiology 133:377\\u0026ndash;392. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1097/ALN.0000000000003347\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1097/ALN.0000000000003347\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBronzino JD, Stern WC, Leahy JP, Morgane PJ (1976) Sleep cycles in cats during chronic electrical stimulation of the area postrema and the anterior raphe. Brain Res Bull 1:235\\u0026ndash;239. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/0361-9230(76)90073-3\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/0361-9230(76)90073-3\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eJiang-Xie LF, Yin L, Zhao S, Prevosto V, Han BX, Dzirasa K, Wang F (2019) A Common Neuroendocrine Substrate for Diverse General Anesthetics and Sleep. Neuron 102:1053\\u0026ndash;1065e1054. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.neuron.2019.03.033\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.neuron.2019.03.033\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLemke SM, Ramanathan DS, Darevksy D, Egert D, Berke JD, Ganguly K (2021) Coupling between motor cortex and striatum increases during sleep over long-term skill learning. Elife 10. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.7554/eLife.64303\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.7554/eLife.64303\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLi H (2023) A preliminary study of brain response region in mice after intraperitoneal injection of dexmedetomidine. Master, Anhui Medical University\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eTaylor NE, Van Dort CJ, Kenny JD, Pei J, Guidera JA, Vlasov KY, Lee JT, Boyden ES, Brown EN, Solt K (2016) Optogenetic activation of dopamine neurons in the ventral tegmental area induces reanimation from general anesthesia. Proc Natl Acad Sci U S A 113:12826\\u0026ndash;12831. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1073/pnas.1614340113\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1073/pnas.1614340113\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChen CR, Zhong YH, Jiang S, Xu W, Xiao L, Wang Z, Qu WM, Huang ZL (2021) Dysfunctions of the paraventricular hypothalamic nucleus induce hypersomnia in mice. Elife 10. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.7554/eLife.69909\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.7554/eLife.69909\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eXu S, Yang H, Menon V, Lemire AL, Wang L, Henry FE, Turaga SC, Sternson SM (2020) Behavioral state coding by molecularly defined paraventricular hypothalamic cell type ensembles. Science 370. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1126/science.abb2494\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1126/science.abb2494\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eYang W, Li H, Cheng Z, Lu Y, Li W, Feng J, Wang L, Cheng J (2022) Dex modulates the balance of water-electrolyte metabolism by depressing the expression of AVP in PVN. Front Pharmacol 13:919032. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.3389/fphar.2022.919032\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.3389/fphar.2022.919032\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eIslam MT, Rumpf F, Tsuno Y, Kodani S, Sakurai T, Matsui A, Maejima T, Mieda M (2022) Vasopressin neurons in the paraventricular hypothalamus promote wakefulness via lateral hypothalamic orexin neurons. Curr Biol 32:3871\\u0026ndash;3885e3874. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.cub.2022.07.020\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.cub.2022.07.020\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eJurek B, Neumann ID (2018) The Oxytocin Receptor: From Intracellular Signaling to Behavior. Physiol Rev 98:1805\\u0026ndash;1908. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1152/physrev.00031.2017\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1152/physrev.00031.2017\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePaxinos G, Franklin KBJ (2004) The mouse brain in stereotaxic coordinates. Elsevier Academic, Amsterdam; Boston\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGe C, Chen W, Zhang L, Ai Y, Zou Y, Peng Q (2023) Chemogenetic activation of the HPC-mPFC pathway improves cognitive dysfunction in lipopolysaccharide -induced brain injury. Theranostics 13:2946\\u0026ndash;2961. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.7150/thno.82889\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.7150/thno.82889\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eTang HD, Dong WY, Hu R, Huang JY, Huang ZH, Xiong W, Xue T, Liu J, Yu JM, Zhu X, Zhang Z (2022) A neural circuit for the suppression of feeding under persistent pain. Nat Metab 4:1746\\u0026ndash;1755. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1038/s42255-022-00688-5\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1038/s42255-022-00688-5\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eFan S, Cheng X, Zhang P, Wang Y, Wang L, Cheng J (2023) The α(2) Adrenoceptor Agonist and Sedative/Anaesthetic Dexmedetomidine Excites Diverse Neuronal Types in the Ventrolateral Preoptic Area of Male Mice. ASN Neuro 15:17590914231191016. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1177/17590914231191016\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1177/17590914231191016\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSu X, Meng Z-T, Wu X-H, Cui F, Li H-L, Wang D-X, Zhu X, Zhu S-N, Maze M, Ma D (2016) Dexmedetomidine for prevention of delirium in elderly patients after non-cardiac surgery: a randomised, double-blind, placebo-controlled trial. Lancet 388:1893\\u0026ndash;1902. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/s0140-6736(16)30580-3\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/s0140-6736(16)30580-3\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChouchou F, Khoury S, Chauny JM, Denis R, Lavigne GJ (2014) Postoperative sleep disruptions: a potential catalyst of acute pain? Sleep Med Rev 18:273\\u0026ndash;282. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.smrv.2013.07.002\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.smrv.2013.07.002\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSong B, Li Y, Teng X, Li X, Yang Y, Zhu J (2019) The Effect Of Intraoperative Use Of Dexmedetomidine During The Daytime Operation Vs The Nighttime Operation On Postoperative Sleep Quality And Pain Under General Anesthesia. Nat Sci Sleep 11:207\\u0026ndash;215. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.2147/nss.S225041\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.2147/nss.S225041\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCarr ZJ, Cios TJ, Potter KF, Swick JT (2018) Does Dexmedetomidine Ameliorate Postoperative Cognitive Dysfunction? A Brief Review of the Recent Literature. Curr Neurol Neurosci Rep 18:64. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1007/s11910-018-0873-z\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1007/s11910-018-0873-z\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eG\\u0026ouml;genur I, Bisgaard T, Burgdorf S, van Someren E, Rosenberg J (2009) Disturbances in the circadian pattern of activity and sleep after laparoscopic versus open abdominal surgery. Surg Endosc 23:1026\\u0026ndash;1031. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1007/s00464-008-0112-9\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1007/s00464-008-0112-9\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNi P, Dong H, Zhou Q, Wang Y, Sun M, Qian Y, Sun J (2019) Preoperative Sleep Disturbance Exaggerates Surgery-Induced Neuroinflammation and Neuronal Damage in Aged Mice. Mediators Inflamm 2019:8301725. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1155/2019/8301725\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1155/2019/8301725\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eRuiz FS, Andersen ML, Guindalini C, Araujo LP, Lopes JD, Tufik S (2017) Sleep influences the immune response and the rejection process alters sleep pattern: Evidence from a skin allograft model in mice. Brain Behav Immun 61:274\\u0026ndash;288. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.bbi.2016.12.027\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.bbi.2016.12.027\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eTabassum S, Misrani A, Tabassum S, Ahmed A, Yang L, Long C (2021) Disrupted prefrontal neuronal oscillations and morphology induced by sleep deprivation in young APP/PS1 transgenic AD mice. Brain Res Bull 166:12\\u0026ndash;20. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.brainresbull.2020.11.003\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.brainresbull.2020.11.003\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eAlthammer F, Grinevich V (2017) Diversity of oxytocin neurons: beyond magno- and parvocellular cell types? J Neuroendocrinol. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1111/jne.12549\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1111/jne.12549\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSwaab DF, Nijveldt F, Pool CW (1975) Distribution of oxytocin and vasopressin in the rat supraoptic and paraventricular nucleus. J Endocrinol 67:461\\u0026ndash;462. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1677/joe.0.0670461\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1677/joe.0.0670461\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBeurel E, Nemeroff CB (2014) Interaction of stress, corticotropin-releasing factor, arginine vasopressin and behaviour. Curr Top Behav Neurosci 18:67\\u0026ndash;80. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1007/7854_2014_306\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1007/7854_2014_306\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKnobloch HS, Charlet A, Hoffmann LC, Eliava M, Khrulev S, Cetin AH, Osten P, Schwarz MK, Seeburg PH, Stoop R, Grinevich V (2012) Evoked axonal oxytocin release in the central amygdala attenuates fear response. Neuron 73:553\\u0026ndash;566. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.neuron.2011.11.030\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.neuron.2011.11.030\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNeumann ID, Landgraf R (2012) Balance of brain oxytocin and vasopressin: implications for anxiety, depression, and social behaviors. Trends Neurosci 35:649\\u0026ndash;659. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/j.tins.2012.08.004\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/j.tins.2012.08.004\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBraga RI, Panaitescu A, Bădescu S, Zăgrean A-M, Zăgrean L (2014) Intranasal administration of oxytocin alters sleep architecture. Biol Rhythm Res 45:69\\u0026ndash;75. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1080/09291016.2013.797641\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1080/09291016.2013.797641\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLancel M, Kr\\u0026ouml;mer S, Neumann ID (2003) Intracerebral oxytocin modulates sleep-wake behaviour in male rats. Regul Pept 114:145\\u0026ndash;152. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1016/s0167-0115(03)00118-6\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1016/s0167-0115(03)00118-6\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMahoney C, Kroeger D, Grinevich V, Scammell T (2017) 0134 Oxytocin fibers in the lateral hypothalamus promote arousal in a mouse model of PWS. Sleep 40:A50\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eVenn RM, Bradshaw CJ, Spencer R, Brealey D, Caudwell E, Naughton C, Vedio A, Singer M, Feneck R, Treacher D, Willatts SM, Grounds RM (1999) Preliminary UK experience of dexmedetomidine, a novel agent for postoperative sedation in the intensive care unit. Anaesthesia 54:1136\\u0026ndash;1142. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1046/j.1365-2044.1999.01114.x\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1046/j.1365-2044.1999.01114.x\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNelson LE, Lu J, Guo T, Saper CB, Franks NP, Maze M (2003) The alpha2-adrenoceptor agonist dexmedetomidine converges on an endogenous sleep-promoting pathway to exert its sedative effects. Anesthesiology 98:428\\u0026ndash;436. \\u003cspan class=\\\"ExternalRef\\\"\\u003e\\u003cspan class=\\\"RefSource\\\"\\u003ehttps://doi.org:10.1097/00000542-200302000-00024\\u003c/span\\u003e\\u003cspan address=\\\"https://doi.org:10.1097/00000542-200302000-00024\\\" targettype=\\\"DOI\\\" class=\\\"RefTarget\\\"\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"neurochemical-research\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"nere\",\"sideBox\":\"Learn more about [Neurochemical Research](https://www.springer.com/journal/11064)\",\"snPcode\":\"11064\",\"submissionUrl\":\"https://submission.nature.com/new-submission/11064/3\",\"title\":\"Neurochemical Research\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false},\"keywords\":\"DEX, OXT neurons, PVN, EEG/EMG recording, patch clamp\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4207174/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4207174/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eThis study aimed to investigate the mechanism of Dexmedetomidine (DEX) on sleep homeostasis. We will focus on the effect of DEX on the activity and the signal transmission of oxytocin-expressing (Oxytocin, OXT) neurons in the paraventricular nucleus (PVN) of the hypothalamus (PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e) in the regulation of sleep-wakefulness cycle. In this study, the guide cannula was implanted to microinject DEX into PVN. PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons were specifically activated by the chemogenetic method. Differences in sleep-wakefulness states were monitored by electroencephalogram (EEG)/ electromyogram (EMG) recording. In addition, the effect of DEX on the electrophysiological activity of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons was examined by whole-cell patch-clamp technique. EEG/EMG results showed that microinjection of DEX in PVN significantly increased the duration of nonrapid eye movement (NREM) sleep in mice. Chemogenetic activation of PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons after internal cannula injection of DEX to PVN increased the amount of wake. Electrophysiological results show that DEX could inhibit the frequency of action potential (AP) and the spontaneous excitatory postsynaptic current (sEPSC) in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors. DEX maintained sleep homeostasis by inhibiting excitatory synaptic signaling in PVN\\u003csup\\u003eOXT\\u003c/sup\\u003e neurons through α\\u003csub\\u003e2\\u003c/sub\\u003e-adrenoceptors.\\u003c/p\\u003e\",\"manuscriptTitle\":\"Dexmedetomidine promotes NREM sleep by depressing the neuronal activity of OXT neurons in the PVN\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-04-08 06:13:48\",\"doi\":\"10.21203/rs.3.rs-4207174/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2024-07-09T08:20:40+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2024-05-13T16:52:23+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"16217354039560399933358161372737475956\",\"date\":\"2024-04-29T18:13:01+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2024-04-07T15:46:51+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2024-04-04T18:06:00+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2024-04-03T02:22:52+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"Neurochemical Research\",\"date\":\"2024-04-02T13:50:56+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"neurochemical-research\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"nere\",\"sideBox\":\"Learn more about [Neurochemical Research](https://www.springer.com/journal/11064)\",\"snPcode\":\"11064\",\"submissionUrl\":\"https://submission.nature.com/new-submission/11064/3\",\"title\":\"Neurochemical Research\",\"twitterHandle\":\"\",\"acdcEnabled\":true,\"dfaEnabled\":true,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"Springer Hybrid\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":false}}],\"origin\":\"\",\"ownerIdentity\":\"6a3050b2-2352-48ac-9a62-348c6e4bbe79\",\"owner\":[],\"postedDate\":\"April 8th, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-08-05T16:00:14+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-4207174\",\"link\":\"https://doi.org/10.1007/s11064-024-04221-w\",\"journal\":{\"identity\":\"neurochemical-research\",\"isVorOnly\":false,\"title\":\"Neurochemical Research\"},\"publishedOn\":\"2024-07-30 15:57:08\",\"publishedOnDateReadable\":\"July 30th, 2024\"},\"versionCreatedAt\":\"2024-04-08 06:13:48\",\"video\":\"\",\"vorDoi\":\"10.1007/s11064-024-04221-w\",\"vorDoiUrl\":\"https://doi.org/10.1007/s11064-024-04221-w\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4207174\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4207174\",\"identity\":\"rs-4207174\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}