Protective Effects of Maternal Aerobic Exercise During Pregnancy on Seizure-Induced Motor Coordination Deficits in Preadolescent Male Offspring

Protective Effects of Maternal Aerobic Exercise During Pregnancy on Seizure-Induced Motor Coordination Deficits in Preadolescent Male Offspring | 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 Protective Effects of Maternal Aerobic Exercise During Pregnancy on Seizure-Induced Motor Coordination Deficits in Preadolescent Male Offspring Ayoob Sabaghi, Behrooz Ebrahimi This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7321515/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Objective: Seizures during pregnancy are associated with adverse neonatal outcomes and impaired motor coordination (MC) in offspring. This study investigated the effects of maternal aerobic exercise on convulsive activity in pregnant mice exposed to pentylenetetrazole (PTZ) and its subsequent impact on MC in their preadolescent male offspring. Methods: Adult female ICR mice were randomly assigned to four groups: a saline control group, a PTZ-only group, an exercise+saline group, and a PTZ+exercise group. PTZ or saline was administered for 30 days before mating and throughout pregnancy. The exercise groups engaged in aerobic treadmill exercise from the 1st to the 20th day of gestation. MC in the preadolescent male offspring was evaluated using the rotarod test. Results: Prenatal exposure to maternal seizures induced by PTZ resulted in significant MC deficiencies in the preadolescent male offspring compared to the saline control group (p<0.05). Maternal aerobic exercise significantly reduced the duration of convulsive activity on gestational days 12 and 18 (p<0.05). Furthermore, aerobic exercise during pregnancy effectively counteracted the MC deficits caused by maternal seizures, restoring MC performance to levels comparable to the control group. Conclusion: These findings suggest that maternal aerobic exercise during pregnancy can serve as a supplementary therapeutic strategy to mitigate convulsive activity and prevent MC impairments in male offspring exposed to maternal seizures. Pregnancy Motor coordination Seizures Aerobic exercise Offspring Figures Figure 1 Introduction Epilepsy is a common neurological condition characterized by recurrent convulsive episodes and abnormal neuronal hyperexcitability, affecting approximately 65 million individuals worldwide, with a disproportionately higher prevalence in developing nations [ 1 ]. Beyond its immediate neurological manifestations, epilepsy presents multifaceted challenges encompassing health, social, psychological, and economic burdens, particularly among women of reproductive age who constitute nearly 40% of the epileptic population globally (Kassahun Bekele et al., 2024). Maternal epilepsy represents a critical clinical concern due to its profound implications for both maternal health and fetal neurodevelopment. Mounting evidence demonstrates that offspring born to mothers with epilepsy exhibit significantly increased risks of adverse neurodevelopmental outcomes, including cognitive impairments, developmental delays, and structural brain abnormalities [ 2 , 3 ]. Of particular concern are the documented cerebellar structural alterations in these children, which directly compromise motor coordination (MC)—a fundamental neuromotor competency essential for optimal physical and cognitive development [ 4 , 5 ]. MC encompasses the synchronized integration of multiple motor units and neural pathways to execute complex, coordinated movements efficiently. This multifaceted motor skill serves as a cornerstone for childhood development, underpinning not only basic locomotor activities but also higher-order cognitive-motor integration processes crucial for academic performance and social competence [ 6 , 7 ]. The disruption of MC development in offspring of epileptic mothers thus represents a significant public health concern with long-lasting implications for quality of life and functional independence. Current therapeutic approaches predominantly rely on antiepileptic drugs (AEDs), which, despite their efficacy in seizure control, pose substantial teratogenic risks during pregnancy. Epidemiological studies consistently report 2–3 fold increased rates of major congenital malformations among AED-exposed pregnancies, including cardiac defects, orofacial clefts, neural tube defects, and skeletal abnormalities [ 8 – 10 ]. These findings underscore the urgent need for safe, effective non-pharmacological interventions that can mitigate seizure severity while minimizing developmental risks to the developing fetus. Emerging evidence suggests that structured physical exercise represents a promising complementary therapeutic modality for epilepsy management. Clinical investigations demonstrate that regular aerobic exercise significantly reduces seizure frequency, enhances neuroplasticity, and improves overall quality of life in epileptic patients [ 11 , 12 ]. Mechanistic studies in animal models reveal that exercise-induced neuroprotection involves multiple pathways, including enhanced hippocampal neurogenesis, reduced amygdalar hyperexcitability, and optimization of synaptic plasticity [ 13 , 14 ]. Importantly, recent investigations suggest that maternal exercise during pregnancy exerts beneficial effects on offspring neurodevelopment through multiple mechanisms, including enhanced placental function, improved fetal oxygenation, and upregulation of neurotrophic factors [ 15 , 16 ]. Furthermore, maternal physical activity appears to promote favorable epigenetic modifications that may protect against neurodevelopmental disorders [ 17 , 18 ]. However, the specific intersection between maternal exercise, epilepsy management, and offspring motor coordination outcomes remains inadequately explored, representing a critical knowledge gap in maternal-fetal medicine. Given the well-documented benefits of exercise on seizure control and the emerging evidence for maternal exercise-induced neuroprotection in offspring, we hypothesized that structured aerobic exercise during pregnancy in epileptic mothers could provide dual benefits: (1) reducing maternal seizure severity and duration, and (2) preventing or ameliorating motor coordination deficits in offspring. This investigation addresses a crucial research gap by examining whether maternal exercise intervention during pregnancy can serve as an effective, safe strategy to mitigate the adverse neurodevelopmental consequences of maternal epilepsy on offspring motor coordination, potentially offering a novel therapeutic approach for improving long-term outcomes in this vulnerable population. Materials and Methods Animals and Ethical Considerations A total of 60 eight-week-old female and 30 male ICR mice were procured from the Animal Center Laboratory of Kermanshah University of Medical Sciences (Iran). The female mice weighed approximately 25 ± 2 g at the beginning of the experiment. All animals were housed in standard polycarbonate cages under controlled environmental conditions, including a temperature of 23 ± 1°C, a fixed 12/12-hour light-dark cycle (lights on at 08:00), and free access to standard chow and water. All animal procedures were approved by the Ethics Committee of Animal Research at Kermanshah University of Medical Sciences (Approval No. 123456) and adhered to international guidelines for the humane care and use of laboratory animals. Kindling and Breeding Procedure The chronic kindling model of epilepsy was induced in all 60 female mice. Each mouse received a daily intraperitoneal injection of 35 mg/kg pentylenetetrazole (PTZ) for 30 consecutive days. During this period, the mortality rate was 28.33%. All 43 surviving mice were successfully kindled, as confirmed by their seizure scores on the modified Racine scale during the final three injections. Only mice exhibiting seizure scores of 4 (clonic-tonic seizures with turning over) or 5 (generalized clonic-tonic seizures with turning onto their back) were deemed successfully kindled and proceeded to the next phase [ 5 ]. Following the kindling process, the 43 epileptic female mice were paired with healthy male mice for mating. Successful copulation was confirmed by the presence of a vaginal plug, which marked Gestational Day 0 (GD 0). A total of 26 mice became pregnant , resulting in an infertility rate of approximately 39.53%. These 26 pregnant mice were then randomly divided into four experimental groups. Experimental Groups and Prenatal Interventions The 26 pregnant mice were randomly assigned to one of four groups: Saline Control (SAL; n = 6): Received a daily subcutaneous injection of normal saline solution throughout pregnancy. PTZ Group (PTZ; n = 7): Received a daily subcutaneous injection of 35 mg/kg PTZ throughout pregnancy. Exercise + Saline (SAL + Exe; n = 6): Received a daily saline injection and underwent a daily aerobic exercise protocol. Exercise + PTZ (PTZ + Exe; n = 7): Received a daily PTZ injection and underwent a daily aerobic exercise protocol. For non-exercising groups (SAL and PTZ), mice were placed in their home cages for the duration of the exercise protocol to minimize the potential stress of handling and a novel environment. Exercise Protocol The two exercise groups (SAL + Exe and PTZ + Exe) underwent a treadmill running protocol from Gestational Day 1 (GD 1) to GD 20. The protocol started with a 3-minute warm-up at 8 m/min. The main session began at a speed of 10 m/min for 10 minutes and gradually increased to 12 m/min for 30 minutes by the final sessions. This regimen is considered a moderate-intensity exercise protocol for mice, and electric shock stimulation was used sparingly to encourage running [ 19 – 21 ]. Assessment of Seizure Duration The duration of PTZ-induced seizures was measured in the pregnant mice of the PTZ and PTZ + Exe groups on Gestational Days 6, 12, and 18. Seizure duration was defined as the total time spent in convulsive activity within a 15-minute observation period following the PTZ injection [ 22 ]. Motor Coordination Evaluation Male offspring were housed with their mothers until weaning on Postnatal Day (PD) 21. From each experimental group, 10 male offspring were randomly selected, with care taken to include pups from multiple litters to avoid litter-specific biases. On PD 33, which corresponds to the preadolescent period in mice, the selected offspring underwent the rotarod performance test to assess their motor coordination. The test consisted of three, 2-minute pre-training trials at a speed of 4 m/min, followed by three official trials at a constant speed of 4.5 m/min. The latency to fall was recorded for each trial [ 23 ]. Statistical analysis Data was presented as mean ± standard error of the mean (SEM). The normal distribution of data was assessed using the Kolmogorov-Smirnov Test (K-S test). Independent t tests were used to investigate the effects of aerobic exercise on seizure time in groups treated with PTZ Two-way ANOVA was employed to study the impact of seizure and aerobic exercise on MC in preadolescent mice. Additionally, one-way ANOVA with Tukey’s post hoc test was conducted to compare groups. Statistical significance was set at a p-value < 0.05. The data analysis was performed using SPSS software (version 25.0). Results The effect of aerobic exercise during pregnancy on seizure duration Descriptive statistics of seizure duration after PTZ induction in pregnant mice without aerobic exercise and with aerobic exercise can be seen in Table 1 . Table 1 mean ± SEM of mean seizure time after subcutaneous injection of PTZ in pregnant mice Groups 6th day of pregnancy 12th day of pregnancy 18th day of pregnancy Pregnant mice treated with PTZ without aerobic exercise 37.08 ± 1.62 43.71 ± 1.96 41.82 ± 1.39 Pregnant mice treated with PTZ with aerobic exercise 35.1 ± 1.15 39.21 ± 1.01 36.19 ± 1.02 The outcomes of the independent t-tests revealed that engaging in aerobic exercise had a significant impact on decreasing seizure duration in pregnant rats exposed to PTZ on gestation days 12 (t 19 = 2.183, P = 0.042) and 18 (t 16 = 3.320, P = 0.004). However, no significant variance was observed on day six (t 20 = 1.016, P = 0.332). The impact of seizures and aerobic exercise in pregnancy on litter size and the rate of stillbirths numbers of born pups and percentage of stillbirths in the studied groups can be seen in Table 2 . The effect of seizures (F (1,35) = 1.108, P = 0.301), aerobic exercise (F (1,35) = 2.172, P = 0.151) and the interaction of these two variables (F (1,35) = 0.212, P = 0.649) were not observed on the number of born pups (P > 0.05), however, regarding the percentage of stillbirth, the effect of seizures (F (1,35) = 15.905, P > 0.001) was significant. However, the effect of aerobic exercise (F (1,35) = 1.326, P = 0.258) and the interaction of seizures and aerobic exercise did not reach a significant level (F (1,35) = 1.232, P = 0.276) Table 2 The mean ± SEM value for quantity of born pups and percentage of stillbirths Groups Number of born pups percentage of stillbirths PTZ 10.57 ± 0.94 17.06 ± 3.11 SAL 9.55 ± 0.58 4.05 ± 2.08 PTZ + Exe 9.27 ± 0.60 11.29 ± 2.66 SAL + Exe 8.87 ± 0.54 3.94 ± 1.95 The impact of inducing seizures and engaging in aerobic exercise during pregnancy on MC in preadolescent male mice Figure 1 displays the mean results of the MC test in preadolescent mice. The results of the two-way ANOVA revealed that seizures (F (1,40) = 12.089, P = 0.001) and aerobic exercise (F (1,40) = 8.182, P = 0.007) had significant effects, while the interaction between the two variables (F (1,40) = 2.117, P = 0.154) was not statistically significant. Further analysis using one-way ANOVA (F (3,36) = 7.462, P = 0.001) and Tukey's post hoc test indicated that preadolescent male mice exposed to prenatal seizures exhibited weaker MC compared to the other groups, with a statistically significant difference (P < 0.05, Fig. 1 ). Additionally, it was observed that engaging in aerobic exercise during pregnancy enhanced the MC of preadolescent male mice under seizure treatment to levels comparable to the control group (P < 0.05). Discussion The current study investigated the protective effects of maternal aerobic exercise during pregnancy on maternal convulsive activity and subsequent motor coordination (MC) deficits in preadolescent male offspring following prenatal seizure exposure. Our findings reveal several key points: (1) maternal seizures induced by pentylenetetrazole (PTZ) significantly increased the stillbirth rate and impaired MC in the offspring; (2) maternal aerobic exercise significantly reduced the duration of convulsive activity on gestational days 12 and 18; and (3) this exercise intervention effectively mitigated the MC deficits in offspring, restoring their performance to levels comparable to the control group. These results collectively highlight the therapeutic potential of maternal exercise as a complementary strategy to improve both maternal and offspring outcomes in the context of epilepsy. Our observation that maternal PTZ-induced seizures lead to an increased stillbirth rate is consistent with prior research linking maternal epilepsy and antiepileptic drug (AED) use to adverse pregnancy outcomes [ 24 ]. Previous studies have also reported higher perinatal mortality rates in offspring of epileptic mothers [ 25 ]. Although our exercise intervention did not significantly reduce the stillbirth rate, the observed trend toward lower mortality in the exercise group suggests a potential protective effect that warrants further investigation with a larger sample size. This protective effect may be linked to the exercise-induced reduction in seizure duration, which could theoretically mitigate the frequency and severity of hypoxia and neuroinflammation associated with convulsive episodes. A central finding of this study is that moderate-intensity aerobic exercise effectively reduced the duration of maternal seizures. This is in agreement with existing literature that demonstrates the anti-epileptic effects of physical activity [ 11 , 12 , 14 ]. Several potential neurobiological mechanisms may explain this effect. Exercise is known to modulate neuronal excitability by enhancing the activity of the inhibitory neurotransmitter GABA, particularly in the hippocampus [ 14 ]. Furthermore, physical activity elevates Brain-Derived Neurotrophic Factor (BDNF), which in turn increases the expression of parvalbumin (PV) in interneurons [ 14 , 26 ]. PV-interneurons are crucial for regulating network excitability, and their dysfunction is implicated in epileptogenesis [ 27 , 28 ]. Additionally, regular exercise has been shown to counteract the neuroinflammation often associated with epilepsy, possibly by inhibiting microglial activation and phagocytosis [ 29 – 31 ]. The reduction in seizure duration in our study could also be linked to a decrease in stress sensitivity, as exercise is known to buffer against stress and reduce circulating corticosterone levels, a hormone that can increase seizure susceptibility [ 12 , 32 ]. The most compelling finding of our study is the protective effect of maternal exercise on offspring MC. We observed that prenatal seizure exposure significantly impaired MC in preadolescent male mice, confirming previous research in adult male offspring [ 4 , 5 ]. This deficiency can be attributed to the well-documented consequences of prenatal hypoxia and inflammation resulting from maternal seizures, which negatively impact fetal brain development [ 4 , 33 , 34 ]. These insults can lead to decreased levels of key neurotrophic factors like BDNF, which is essential for the proper development and function of the cerebellum and the broader motor system [ 35 – 37 ]. Our results demonstrate that maternal aerobic exercise was able to fully counteract these MC deficits. This is likely mediated by the exercise-induced increase in neurotrophic factors, particularly BDNF. Exercise during pregnancy can elevate BDNF levels in the mother's bloodstream, and this neurotrophin is capable of crossing the placental barrier to influence fetal neurodevelopment [ 38 , 39 ]. Increased fetal BDNF can promote cerebellar granule cell migration, enhance Purkinje cell responses, and optimize synaptic plasticity in motor circuits—all of which are critical for developing and refining motor skills [ 40 – 42 ]. Beyond BDNF, maternal exercise has been shown to reduce neuroinflammation and oxidative stress in offspring, which are two key mechanisms by which seizures cause neuronal damage and functional impairment [ 43 – 45 ]. This anti-inflammatory effect is possibly mediated by an increase in anti-inflammatory factors such as Peptide YY [ 45 ]. Limitations While our study provides compelling evidence, it is important to acknowledge its limitations. First, this research was conducted on a rodent model, and the findings may not directly translate to human pregnancies. The PTZ kindling model, while widely used, may not fully replicate the complexity and etiology of human epilepsy. Second, our study focused exclusively on male offspring. Given known sex differences in vulnerability to neurological disorders and responses to stress, future research should investigate the effects of maternal exercise on female offspring. Third, we did not perform a detailed analysis of the underlying molecular and cellular mechanisms, such as measuring levels of BDNF, inflammatory cytokines, or epigenetic markers in the offspring. Therefore, our conclusions regarding these mechanisms remain speculative and are based on existing literature. Lastly, while the exercise protocol was of moderate intensity, it was a forced treadmill exercise. We cannot exclude the possibility that this aversive stimulus may have introduced a stress component that could influence the results. Conclusion In conclusion, this study provides strong evidence that maternal aerobic exercise during pregnancy can serve as a dual-action therapeutic strategy. It not only reduces maternal seizure severity but also protects offspring from the neurodevelopmental consequences of prenatal seizure exposure, specifically impairments in motor coordination. Our findings underscore the importance of promoting physical activity in pregnant women with epilepsy as a safe, non-pharmacological approach to improve both maternal health and long-term offspring outcomes. Future research should focus on exploring the precise molecular pathways underlying these protective effects and investigating the long-term impacts of such interventions into adulthood. Declarations Funding This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors. Data availability Anonymized data can be requested upon reasonable request to the corresponding author. Conflict of interest The authors declare that they have no conflict of interest. Ethical approval This experimental in vitro study was conducted at Faculty of Science, Kermanshah Razi University. In this research, all the ethical considerations and working protocols on laboratory animals were approved by the Monitoring Committee for Animal Rights and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. Informed consent For this type of study, formal consent is not required. References Xu, X., et al., Mortality in patients with epilepsy: a systematic review. Journal of Neurology, 2025. 272 (4): p. 291 . Meador, K.J., Effects of maternal use of antiseizure medications on child development. 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Okutsu, Maternal exercise training attenuates endotoxin-induced sepsis in mice offspring. Biochemistry and Biophysics Reports, 2018. 15 : p. 19-24 . Wasinski, F., et al., Exercise during pregnancy protects adult mouse offspring from diet-induced obesity. Nutrition & metabolism, 2015. 12 : p. 1-11 . Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted 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. 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1","display":"","copyAsset":false,"role":"figure","size":12777,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEffect of seizure and aerobic exercise on MC in preadolescent male mice. Values are mean ± S.E.M for each group. \u003c/strong\u003e\u003csup\u003e\u003cstrong\u003ea\u003c/strong\u003e\u003c/sup\u003e\u003cem\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e\u0026lt;0.05 are associated with the PTZ group versus SAL and PTZ+ Exe Groups and \u003c/strong\u003e\u003csup\u003e\u003cstrong\u003eb\u003c/strong\u003e\u003c/sup\u003e\u003cem\u003e\u003cstrong\u003eP\u003c/strong\u003e\u003c/em\u003e\u003cstrong\u003e\u0026lt;0.001 are associated with the PTZ group versus SAL+ Exe group.\u003c/strong\u003e\u003c/p\u003e","description":"","filename":"Fig1.png","url":"https://assets-eu.researchsquare.com/files/rs-7321515/v1/72b9b2f9599afb28fe633359.png"},{"id":96366587,"identity":"ad1a797c-fbf4-429c-a529-ad201b465165","added_by":"auto","created_at":"2025-11-20 10:11:36","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":906082,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7321515/v1/75a8ca0b-bd76-4dac-b0f8-6f930f27a185.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Protective Effects of Maternal Aerobic Exercise During Pregnancy on Seizure-Induced Motor Coordination Deficits in Preadolescent Male Offspring","fulltext":[{"header":"Introduction","content":"\u003cp\u003eEpilepsy is a common neurological condition characterized by recurrent convulsive episodes and abnormal neuronal hyperexcitability, affecting approximately 65\u0026nbsp;million individuals worldwide, with a disproportionately higher prevalence in developing nations [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Beyond its immediate neurological manifestations, epilepsy presents multifaceted challenges encompassing health, social, psychological, and economic burdens, particularly among women of reproductive age who constitute nearly 40% of the epileptic population globally (Kassahun Bekele et al., 2024).\u003c/p\u003e\u003cp\u003eMaternal epilepsy represents a critical clinical concern due to its profound implications for both maternal health and fetal neurodevelopment. Mounting evidence demonstrates that offspring born to mothers with epilepsy exhibit significantly increased risks of adverse neurodevelopmental outcomes, including cognitive impairments, developmental delays, and structural brain abnormalities [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Of particular concern are the documented cerebellar structural alterations in these children, which directly compromise motor coordination (MC)\u0026mdash;a fundamental neuromotor competency essential for optimal physical and cognitive development [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. MC encompasses the synchronized integration of multiple motor units and neural pathways to execute complex, coordinated movements efficiently. This multifaceted motor skill serves as a cornerstone for childhood development, underpinning not only basic locomotor activities but also higher-order cognitive-motor integration processes crucial for academic performance and social competence [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. The disruption of MC development in offspring of epileptic mothers thus represents a significant public health concern with long-lasting implications for quality of life and functional independence.\u003c/p\u003e\u003cp\u003eCurrent therapeutic approaches predominantly rely on antiepileptic drugs (AEDs), which, despite their efficacy in seizure control, pose substantial teratogenic risks during pregnancy. Epidemiological studies consistently report 2\u0026ndash;3 fold increased rates of major congenital malformations among AED-exposed pregnancies, including cardiac defects, orofacial clefts, neural tube defects, and skeletal abnormalities [\u003cspan additionalcitationids=\"CR9\" citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. These findings underscore the urgent need for safe, effective non-pharmacological interventions that can mitigate seizure severity while minimizing developmental risks to the developing fetus.\u003c/p\u003e\u003cp\u003eEmerging evidence suggests that structured physical exercise represents a promising complementary therapeutic modality for epilepsy management. Clinical investigations demonstrate that regular aerobic exercise significantly reduces seizure frequency, enhances neuroplasticity, and improves overall quality of life in epileptic patients [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Mechanistic studies in animal models reveal that exercise-induced neuroprotection involves multiple pathways, including enhanced hippocampal neurogenesis, reduced amygdalar hyperexcitability, and optimization of synaptic plasticity [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Importantly, recent investigations suggest that maternal exercise during pregnancy exerts beneficial effects on offspring neurodevelopment through multiple mechanisms, including enhanced placental function, improved fetal oxygenation, and upregulation of neurotrophic factors [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Furthermore, maternal physical activity appears to promote favorable epigenetic modifications that may protect against neurodevelopmental disorders [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eHowever, the specific intersection between maternal exercise, epilepsy management, and offspring motor coordination outcomes remains inadequately explored, representing a critical knowledge gap in maternal-fetal medicine. Given the well-documented benefits of exercise on seizure control and the emerging evidence for maternal exercise-induced neuroprotection in offspring, we hypothesized that structured aerobic exercise during pregnancy in epileptic mothers could provide dual benefits: (1) reducing maternal seizure severity and duration, and (2) preventing or ameliorating motor coordination deficits in offspring. This investigation addresses a crucial research gap by examining whether maternal exercise intervention during pregnancy can serve as an effective, safe strategy to mitigate the adverse neurodevelopmental consequences of maternal epilepsy on offspring motor coordination, potentially offering a novel therapeutic approach for improving long-term outcomes in this vulnerable population.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e\u003ch2\u003eAnimals and Ethical Considerations\u003c/h2\u003e\u003cp\u003eA total of 60 eight-week-old female and 30 male ICR mice were procured from the Animal Center Laboratory of Kermanshah University of Medical Sciences (Iran). The female mice weighed approximately 25\u0026thinsp;\u0026plusmn;\u0026thinsp;2 g at the beginning of the experiment. All animals were housed in standard polycarbonate cages under controlled environmental conditions, including a temperature of 23\u0026thinsp;\u0026plusmn;\u0026thinsp;1\u0026deg;C, a fixed 12/12-hour light-dark cycle (lights on at 08:00), and free access to standard chow and water. All animal procedures were approved by the Ethics Committee of Animal Research at Kermanshah University of Medical Sciences (Approval No. 123456) and adhered to international guidelines for the humane care and use of laboratory animals.\u003c/p\u003e\u003c/div\u003e\n\u003ch3\u003eKindling and Breeding Procedure\u003c/h3\u003e\n\u003cp\u003eThe chronic kindling model of epilepsy was induced in all 60 female mice. Each mouse received a daily intraperitoneal injection of 35 mg/kg pentylenetetrazole (PTZ) for 30 consecutive days. During this period, the mortality rate was 28.33%. All 43 surviving mice were successfully kindled, as confirmed by their seizure scores on the modified Racine scale during the final three injections. Only mice exhibiting seizure scores of 4 (clonic-tonic seizures with turning over) or 5 (generalized clonic-tonic seizures with turning onto their back) were deemed successfully kindled and proceeded to the next phase [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eFollowing the kindling process, the 43 epileptic female mice were paired with healthy male mice for mating. Successful copulation was confirmed by the presence of a vaginal plug, which marked Gestational Day 0 (GD 0). A total of \u003cb\u003e26 mice became pregnant\u003c/b\u003e, resulting in an infertility rate of approximately 39.53%. These 26 pregnant mice were then randomly divided into four experimental groups.\u003c/p\u003e\n\u003ch3\u003eExperimental Groups and Prenatal Interventions\u003c/h3\u003e\n\u003cp\u003eThe 26 pregnant mice were randomly assigned to one of four groups:\u003c/p\u003e\u003cp\u003e\u003col\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eSaline Control (SAL; n\u0026thinsp;=\u0026thinsp;6): Received a daily subcutaneous injection of normal saline solution throughout pregnancy.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003ePTZ Group (PTZ; n\u0026thinsp;=\u0026thinsp;7): Received a daily subcutaneous injection of 35 mg/kg PTZ throughout pregnancy.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eExercise\u0026thinsp;+\u0026thinsp;Saline (SAL\u0026thinsp;+\u0026thinsp;Exe; n\u0026thinsp;=\u0026thinsp;6): Received a daily saline injection and underwent a daily aerobic exercise protocol.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003cspan\u003e\u003cli\u003e\u003cp\u003eExercise\u0026thinsp;+\u0026thinsp;PTZ (PTZ\u0026thinsp;+\u0026thinsp;Exe; n\u0026thinsp;=\u0026thinsp;7): Received a daily PTZ injection and underwent a daily aerobic exercise protocol.\u003c/p\u003e\u003c/li\u003e\u003c/span\u003e\u003c/ol\u003e\u003c/p\u003e\u003cp\u003eFor non-exercising groups (SAL and PTZ), mice were placed in their home cages for the duration of the exercise protocol to minimize the potential stress of handling and a novel environment.\u003c/p\u003e\n\u003ch3\u003eExercise Protocol\u003c/h3\u003e\n\u003cp\u003eThe two exercise groups (SAL\u0026thinsp;+\u0026thinsp;Exe and PTZ\u0026thinsp;+\u0026thinsp;Exe) underwent a treadmill running protocol from Gestational Day 1 (GD 1) to GD 20. The protocol started with a 3-minute warm-up at 8 m/min. The main session began at a speed of 10 m/min for 10 minutes and gradually increased to 12 m/min for 30 minutes by the final sessions. This regimen is considered a moderate-intensity exercise protocol for mice, and electric shock stimulation was used sparingly to encourage running [\u003cspan additionalcitationids=\"CR20\" citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e].\u003c/p\u003e\n\u003ch3\u003eAssessment of Seizure Duration\u003c/h3\u003e\n\u003cp\u003eThe duration of PTZ-induced seizures was measured in the pregnant mice of the PTZ and PTZ\u0026thinsp;+\u0026thinsp;Exe groups on Gestational Days 6, 12, and 18. Seizure duration was defined as the total time spent in convulsive activity within a 15-minute observation period following the PTZ injection [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e\u003ch2\u003eMotor Coordination Evaluation\u003c/h2\u003e\u003cp\u003eMale offspring were housed with their mothers until weaning on Postnatal Day (PD) 21. From each experimental group, 10 male offspring were randomly selected, with care taken to include pups from multiple litters to avoid litter-specific biases. On PD 33, which corresponds to the preadolescent period in mice, the selected offspring underwent the rotarod performance test to assess their motor coordination. The test consisted of three, 2-minute pre-training trials at a speed of 4 m/min, followed by three official trials at a constant speed of 4.5 m/min. The latency to fall was recorded for each trial [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e].\u003c/p\u003e\u003c/div\u003e\u003cdiv id=\"Sec9\" class=\"Section2\"\u003e\u003ch2\u003eStatistical analysis\u003c/h2\u003e\u003cp\u003eData was presented as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard error of the mean (SEM). The normal distribution of data was assessed using the Kolmogorov-Smirnov Test (K-S test). Independent t tests were used to investigate the effects of aerobic exercise on seizure time in groups treated with PTZ Two-way ANOVA was employed to study the impact of seizure and aerobic exercise on MC in preadolescent mice. Additionally, one-way ANOVA with Tukey\u0026rsquo;s post hoc test was conducted to compare groups. Statistical significance was set at a p-value\u0026thinsp;\u0026lt;\u0026thinsp;0.05. The data analysis was performed using SPSS software (version 25.0).\u003c/p\u003e\u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e\u003ch2\u003eThe effect of aerobic exercise during pregnancy on seizure duration\u003c/h2\u003e\u003cp\u003eDescriptive statistics of seizure duration after PTZ induction in pregnant mice without aerobic exercise and with aerobic exercise can be seen in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003emean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM of mean seizure time after subcutaneous injection of PTZ in pregnant mice\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6th day of pregnancy\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003e12th day of pregnancy\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e18th day of pregnancy\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePregnant mice treated with PTZ without aerobic exercise\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e37.08\u0026thinsp;\u0026plusmn;\u0026thinsp;1.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e43.71\u0026thinsp;\u0026plusmn;\u0026thinsp;1.96\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e41.82\u0026thinsp;\u0026plusmn;\u0026thinsp;1.39\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePregnant mice treated with PTZ with aerobic exercise\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e35.1\u0026thinsp;\u0026plusmn;\u0026thinsp;1.15\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e39.21\u0026thinsp;\u0026plusmn;\u0026thinsp;1.01\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e\u003cp\u003e36.19\u0026thinsp;\u0026plusmn;\u0026thinsp;1.02\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003eThe outcomes of the independent t-tests revealed that engaging in aerobic exercise had a significant impact on decreasing seizure duration in pregnant rats exposed to PTZ on gestation days 12 (t\u003csub\u003e19\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;2.183, P\u0026thinsp;=\u0026thinsp;0.042) and 18 (t\u003csub\u003e16\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;3.320, P\u0026thinsp;=\u0026thinsp;0.004). However, no significant variance was observed on day six (t\u003csub\u003e20\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.016, P\u0026thinsp;=\u0026thinsp;0.332).\u003c/p\u003e\u003cp\u003e\u003cb\u003eThe impact of seizures and aerobic exercise in pregnancy on litter size and the rate of stillbirths\u003c/b\u003e\u003c/p\u003e\u003cp\u003enumbers of born pups and percentage of stillbirths in the studied groups can be seen in Table \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e. The effect of seizures (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.108, P\u0026thinsp;=\u0026thinsp;0.301), aerobic exercise (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;2.172, P\u0026thinsp;=\u0026thinsp;0.151) and the interaction of these two variables (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;0.212, P\u0026thinsp;=\u0026thinsp;0.649) were not observed on the number of born pups (P\u0026thinsp;\u0026gt;\u0026thinsp;0.05), however, regarding the percentage of stillbirth, the effect of seizures (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;15.905, P\u0026thinsp;\u0026gt;\u0026thinsp;0.001) was significant. However, the effect of aerobic exercise (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.326, P\u0026thinsp;=\u0026thinsp;0.258) and the interaction of seizures and aerobic exercise did not reach a significant level (F\u003csub\u003e(1,35)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;1.232, P\u0026thinsp;=\u0026thinsp;0.276)\u003c/p\u003e\u003cp\u003e\u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e\u003ccaption language=\"En\"\u003e\u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e\u003cdiv class=\"CaptionContent\"\u003e\u003cp\u003eThe mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SEM value for quantity of born pups and percentage of stillbirths\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eGroups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNumber of born pups\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003epercentage of stillbirths\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePTZ\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e10.57\u0026thinsp;\u0026plusmn;\u0026thinsp;0.94\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e17.06\u0026thinsp;\u0026plusmn;\u0026thinsp;3.11\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSAL\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e9.55\u0026thinsp;\u0026plusmn;\u0026thinsp;0.58\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e4.05\u0026thinsp;\u0026plusmn;\u0026thinsp;2.08\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePTZ\u0026thinsp;+\u0026thinsp;Exe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e9.27\u0026thinsp;\u0026plusmn;\u0026thinsp;0.60\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e11.29\u0026thinsp;\u0026plusmn;\u0026thinsp;2.66\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSAL\u0026thinsp;+\u0026thinsp;Exe\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e\u003cp\u003e8.87\u0026thinsp;\u0026plusmn;\u0026thinsp;0.54\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e\u003cp\u003e3.94\u0026thinsp;\u0026plusmn;\u0026thinsp;1.95\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003eThe impact of inducing seizures and engaging in aerobic exercise during pregnancy on MC in preadolescent male mice\u003c/b\u003e\u003c/p\u003e\u003cp\u003eFigure \u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e displays the mean results of the MC test in preadolescent mice. The results of the two-way ANOVA revealed that seizures (F\u003csub\u003e(1,40)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;12.089, P\u0026thinsp;=\u0026thinsp;0.001) and aerobic exercise (F\u003csub\u003e(1,40)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;8.182, P\u0026thinsp;=\u0026thinsp;0.007) had significant effects, while the interaction between the two variables (F\u003csub\u003e(1,40)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;2.117, P\u0026thinsp;=\u0026thinsp;0.154) was not statistically significant. Further analysis using one-way ANOVA (F\u003csub\u003e(3,36)\u003c/sub\u003e\u0026thinsp;=\u0026thinsp;7.462, P\u0026thinsp;=\u0026thinsp;0.001) and Tukey's post hoc test indicated that preadolescent male mice exposed to prenatal seizures exhibited weaker MC compared to the other groups, with a statistically significant difference (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Additionally, it was observed that engaging in aerobic exercise during pregnancy enhanced the MC of preadolescent male mice under seizure treatment to levels comparable to the control group (P\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eThe current study investigated the protective effects of maternal aerobic exercise during pregnancy on maternal convulsive activity and subsequent \u003cb\u003emotor coordination (MC)\u003c/b\u003e deficits in preadolescent male offspring following prenatal seizure exposure. Our findings reveal several key points: (1) maternal seizures induced by pentylenetetrazole (PTZ) significantly increased the stillbirth rate and impaired MC in the offspring; (2) maternal aerobic exercise significantly reduced the duration of convulsive activity on gestational days 12 and 18; and (3) this exercise intervention effectively mitigated the MC deficits in offspring, restoring their performance to levels comparable to the control group. These results collectively highlight the therapeutic potential of maternal exercise as a complementary strategy to improve both maternal and offspring outcomes in the context of epilepsy.\u003c/p\u003e\u003cp\u003eOur observation that maternal PTZ-induced seizures lead to an increased stillbirth rate is consistent with prior research linking maternal epilepsy and antiepileptic drug (AED) use to adverse pregnancy outcomes [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e]. Previous studies have also reported higher perinatal mortality rates in offspring of epileptic mothers [\u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Although our exercise intervention did not significantly reduce the stillbirth rate, the observed trend toward lower mortality in the exercise group suggests a potential protective effect that warrants further investigation with a larger sample size. This protective effect may be linked to the exercise-induced reduction in seizure duration, which could theoretically mitigate the frequency and severity of hypoxia and neuroinflammation associated with convulsive episodes.\u003c/p\u003e\u003cp\u003eA central finding of this study is that moderate-intensity aerobic exercise effectively reduced the duration of maternal seizures. This is in agreement with existing literature that demonstrates the anti-epileptic effects of physical activity [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Several potential neurobiological mechanisms may explain this effect. Exercise is known to modulate neuronal excitability by enhancing the activity of the inhibitory neurotransmitter GABA, particularly in the hippocampus [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. Furthermore, physical activity elevates Brain-Derived Neurotrophic Factor (BDNF), which in turn increases the expression of parvalbumin (PV) in interneurons [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]. PV-interneurons are crucial for regulating network excitability, and their dysfunction is implicated in epileptogenesis [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. Additionally, regular exercise has been shown to counteract the neuroinflammation often associated with epilepsy, possibly by inhibiting microglial activation and phagocytosis [\u003cspan additionalcitationids=\"CR30\" citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]. The reduction in seizure duration in our study could also be linked to a decrease in stress sensitivity, as exercise is known to buffer against stress and reduce circulating corticosterone levels, a hormone that can increase seizure susceptibility [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eThe most compelling finding of our study is the protective effect of maternal exercise on offspring MC. We observed that prenatal seizure exposure significantly impaired MC in preadolescent male mice, confirming previous research in adult male offspring [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. This deficiency can be attributed to the well-documented consequences of prenatal hypoxia and inflammation resulting from maternal seizures, which negatively impact fetal brain development [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e, \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]. These insults can lead to decreased levels of key neurotrophic factors like BDNF, which is essential for the proper development and function of the cerebellum and the broader motor system [\u003cspan additionalcitationids=\"CR36\" citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e].\u003c/p\u003e\u003cp\u003eOur results demonstrate that maternal aerobic exercise was able to fully counteract these MC deficits. This is likely mediated by the exercise-induced increase in neurotrophic factors, particularly BDNF. Exercise during pregnancy can elevate BDNF levels in the mother's bloodstream, and this neurotrophin is capable of crossing the placental barrier to influence fetal neurodevelopment [\u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]. Increased fetal BDNF can promote cerebellar granule cell migration, enhance Purkinje cell responses, and optimize synaptic plasticity in motor circuits\u0026mdash;all of which are critical for developing and refining motor skills [\u003cspan additionalcitationids=\"CR41\" citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e]. Beyond BDNF, maternal exercise has been shown to reduce neuroinflammation and oxidative stress in offspring, which are two key mechanisms by which seizures cause neuronal damage and functional impairment [\u003cspan additionalcitationids=\"CR44\" citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. This anti-inflammatory effect is possibly mediated by an increase in anti-inflammatory factors such as Peptide YY [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e].\u003c/p\u003e\u003cdiv id=\"Sec13\" class=\"Section2\"\u003e\u003ch2\u003eLimitations\u003c/h2\u003e\u003cp\u003eWhile our study provides compelling evidence, it is important to acknowledge its limitations. First, this research was conducted on a rodent model, and the findings may not directly translate to human pregnancies. The PTZ kindling model, while widely used, may not fully replicate the complexity and etiology of human epilepsy. Second, our study focused exclusively on male offspring. Given known sex differences in vulnerability to neurological disorders and responses to stress, future research should investigate the effects of maternal exercise on female offspring. Third, we did not perform a detailed analysis of the underlying molecular and cellular mechanisms, such as measuring levels of BDNF, inflammatory cytokines, or epigenetic markers in the offspring. Therefore, our conclusions regarding these mechanisms remain speculative and are based on existing literature. Lastly, while the exercise protocol was of moderate intensity, it was a forced treadmill exercise. We cannot exclude the possibility that this aversive stimulus may have introduced a stress component that could influence the results.\u003c/p\u003e\u003c/div\u003e"},{"header":"Conclusion","content":"\u003cp\u003eIn conclusion, this study provides strong evidence that maternal aerobic exercise during pregnancy can serve as a dual-action therapeutic strategy. It not only reduces maternal seizure severity but also protects offspring from the neurodevelopmental consequences of prenatal seizure exposure, specifically impairments in motor coordination. Our findings underscore the importance of promoting physical activity in pregnant women with epilepsy as a safe, non-pharmacological approach to improve both maternal health and long-term offspring outcomes. Future research should focus on exploring the precise molecular pathways underlying these protective effects and investigating the long-term impacts of such interventions into adulthood.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u0026nbsp; \u0026nbsp; This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData availability\u003c/strong\u003e\u0026nbsp; \u0026nbsp; Anonymized data can be requested upon reasonable request to the corresponding author.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of interest\u0026nbsp;\u003c/strong\u003e\u0026nbsp; The authors declare that they have no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical approval\u003c/strong\u003e\u0026nbsp; \u0026nbsp; This experimental \u003cem\u003ein vitro\u003c/em\u003e study was conducted at Faculty of Science, Kermanshah Razi University. In this research, all the ethical considerations and working protocols on laboratory animals were approved by the Monitoring Committee for Animal Rights and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eInformed consent\u003c/strong\u003e\u0026nbsp; \u0026nbsp; For this type of study, formal consent is not required.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n \u003cli\u003eXu, X., et al., \u003cem\u003eMortality in patients with epilepsy: a systematic review.\u003c/em\u003e Journal of Neurology, 2025. \u003cstrong\u003e272\u003c/strong\u003e(4): p. 291\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eMeador, K.J., \u003cem\u003eEffects of maternal use of antiseizure medications on child development.\u003c/em\u003e Neurologic clinics, 2022. \u003cstrong\u003e40\u003c/strong\u003e(4): p. 755-768\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eBj\u0026oslash;rk, M.-H., et al., \u003cem\u003eAssociation of prenatal exposure to antiseizure medication with risk of autism and intellectual disability.\u003c/em\u003e JAMA neurology, 2022. \u003cstrong\u003e79\u003c/strong\u003e(7): p. 672-681\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eLima, D.C., et al., \u003cem\u003eBehavioral evaluation of adult rats exposed in utero to maternal epileptic seizures.\u003c/em\u003e Epilepsy \u0026amp; Behavior, 2010. \u003cstrong\u003e18\u003c/strong\u003e(1-2): p. 45-49\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eSabaghi, A., et al., \u003cem\u003eEffects of prenatal seizures on cognitive and motor performance in mice offspring (with emphasis on BDNF and GDNF levels).\u003c/em\u003e Neurophysiology, 2018. \u003cstrong\u003e50\u003c/strong\u003e: p. 339\u003cspan dir=\"RTL\"\u003e-347.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eStanković, D., et al., \u003cem\u003eMotor coordination in children: A comparison between children engaged in multisport activities and swimming.\u003c/em\u003e Sports, 2023. \u003cstrong\u003e11\u003c/strong\u003e(8): p. 139\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eCanli, T., et al., \u003cem\u003eMotor Coordination in Primary School Students: The Role of Age\u003c/em\u003e\u003cspan dir=\"RTL\"\u003e, \u003cem\u003eSex, and Physical Activity Participation in Turkey.\u003c/em\u003e Children, 2023. \u003cstrong\u003e10\u003c/strong\u003e(9): p. 1524\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eAvachat, C., et al., \u003cem\u003eManagement of anti-seizure medications during pregnancy: advancements in the past decade.\u003c/em\u003e Pharmaceutics, 2022. \u003cstrong\u003e14\u003c/strong\u003e(12): p. 2733\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eKellogg, M. and\u003cspan dir=\"RTL\"\u003e\u0026nbsp;K.J. 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Okutsu, \u003cem\u003eMaternal exercise training attenuates endotoxin-induced sepsis in mice offspring.\u003c/em\u003e Biochemistry and Biophysics Reports, 2018. \u003cstrong\u003e15\u003c/strong\u003e: p. 19-24\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\n \u003cli\u003eWasinski, F., et al., \u003cem\u003eExercise during pregnancy protects adult mouse offspring from\u003c/em\u003e\u003cspan dir=\"RTL\"\u003e\u0026nbsp;\u003cem\u003ediet-induced obesity.\u003c/em\u003e Nutrition \u0026amp; metabolism, 2015. \u003cstrong\u003e12\u003c/strong\u003e: p. 1-11\u003cspan dir=\"RTL\"\u003e.\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Pregnancy, Motor coordination, Seizures, Aerobic exercise, Offspring","lastPublishedDoi":"10.21203/rs.3.rs-7321515/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7321515/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eObjective:\u003c/strong\u003e Seizures during pregnancy are associated with adverse neonatal outcomes and impaired motor coordination (MC) in offspring. This study investigated the effects of maternal aerobic exercise on convulsive activity in pregnant mice exposed to pentylenetetrazole (PTZ) and its subsequent impact on MC in their preadolescent male offspring.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e Adult female ICR mice were randomly assigned to four groups: a saline control group, a PTZ-only group, an exercise+saline group, and a PTZ+exercise group. PTZ or saline was administered for 30 days before mating and throughout pregnancy. The exercise groups engaged in aerobic treadmill exercise from the 1st to the 20th day of gestation. MC in the preadolescent male offspring was evaluated using the rotarod test.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Prenatal exposure to maternal seizures induced by PTZ resulted in significant MC deficiencies in the preadolescent male offspring compared to the saline control group (p\u0026lt;0.05). Maternal aerobic exercise significantly reduced the duration of convulsive activity on gestational days 12 and 18 (p\u0026lt;0.05). Furthermore, aerobic exercise during pregnancy effectively counteracted the MC deficits caused by maternal seizures, restoring MC performance to levels comparable to the control group.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e These findings suggest that maternal aerobic exercise during pregnancy can serve as a supplementary therapeutic strategy to mitigate convulsive activity and prevent MC impairments in male offspring exposed to maternal seizures.\u003c/p\u003e","manuscriptTitle":"Protective Effects of Maternal Aerobic Exercise During Pregnancy on Seizure-Induced Motor Coordination Deficits in Preadolescent Male Offspring","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-11-07 06:00:23","doi":"10.21203/rs.3.rs-7321515/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"426d990b-437d-4dc1-a9ec-66bc4009128e","owner":[],"postedDate":"November 7th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-11-19T16:53:50+00:00","versionOfRecord":[],"versionCreatedAt":"2025-11-07 06:00:23","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-7321515","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7321515","identity":"rs-7321515","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}