Control of mammalian brain ageing by the unfolded protein response transcription factor XBP1

Control of mammalian brain ageing by the unfolded protein response transcription factor XBP1 | 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 Article Control of mammalian brain ageing by the unfolded protein response transcription factor XBP1 Felipe Cabral-Miranda, Giovanni Tamburini, Gabriela Martinez, and 21 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-83658/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 Brain ageing is the main risk factor to develop dementia and neurodegenerative diseases, associated with a decay in the buffering capacity of the proteostasis network. We investigated the significance of the unfolded protein response (UPR), a major signaling pathway to cope with ER stress, to the functional deterioration of the brain during aging. Genetic disruption of the ER stress sensor IRE1α accelerated cognitive and motor decline during ageing. Exogenous bolstering of the UPR by overexpressing an active form of the UPR transcription factor XBP1 restored synaptic and cognitive function, in addition to reducing cell senescence. Proteomic profiling of hippocampal tissue indicated that XBP1s expression attenuated age-related alterations to synaptic function and pathways linked to neurodegenerative diseases. Overall, our results demonstrate that strategies to manipulate the UPR in mammals may sustain healthy brain ageing. Cognitive Neuroscience Cellular & Molecular Neuroscience Neurodegenerative Disease Proteostatsis Network ER Stress Figures Figure 1 Figure 2 Figure 3 Figure 4 Full Text Additional Declarations Yes there is potential Competing Interest. CH and FCM declare a conflict of interest for a submitted patent application. Inventors: Claudio Hetz, Felipe Cabral Miranda. Title: Treatment of aging or age- related disorders using XBP1. Provisional application for patent at USPTO, application number 62800229. Submitted 01/02/2019. Status: patent pending. Disclosure: PS is a Sanofi employee. 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. 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-83658","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":2853072,"identity":"e956d02a-0447-469c-a11d-8b4c7e75d65a","order_by":0,"name":"Felipe Cabral-Miranda","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Felipe","middleName":"","lastName":"Cabral-Miranda","suffix":""},{"id":2853073,"identity":"298ad3dc-bac2-4fe9-a48a-50242b72761d","order_by":1,"name":"Giovanni Tamburini","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Giovanni","middleName":"","lastName":"Tamburini","suffix":""},{"id":2853074,"identity":"21f63856-2ac1-4895-bb18-38f6a27a7d0e","order_by":2,"name":"Gabriela Martinez","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Gabriela","middleName":"","lastName":"Martinez","suffix":""},{"id":2853075,"identity":"93dff2b0-1f05-4920-a4f9-743152ec6164","order_by":3,"name":"Danilo Medinas","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Danilo","middleName":"","lastName":"Medinas","suffix":""},{"id":2853076,"identity":"0f71a159-3d11-4376-8a53-386090b8c922","order_by":4,"name":"Yannis Gerakis","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Yannis","middleName":"","lastName":"Gerakis","suffix":""},{"id":2853077,"identity":"eff28b1e-c46c-4afc-9bdf-eea935351c8c","order_by":5,"name":"Tim Miedema","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tim","middleName":"","lastName":"Miedema","suffix":""},{"id":2853078,"identity":"a6ee5a39-5a49-4e99-b09a-7d7d94f8f783","order_by":6,"name":"Claudia Duran-Aniotz","email":"","orcid":"","institution":"University of Chile","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Claudia","middleName":"","lastName":"Duran-Aniotz","suffix":""},{"id":2853079,"identity":"fa375f61-0866-4b14-8562-e63b2120ee31","order_by":7,"name":"Alvaro Ardiles","email":"","orcid":"","institution":"University of Valparaiso","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Alvaro","middleName":"","lastName":"Ardiles","suffix":""},{"id":2853080,"identity":"31887dc3-f207-4a8a-95e5-1c25e68bfbc7","order_by":8,"name":"Cristobal Ibaceta Ibaceta-Gonzalez","email":"","orcid":"","institution":"University of Valparaiso","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Cristobal","middleName":"Ibaceta","lastName":"Ibaceta-Gonzalez","suffix":""},{"id":2853081,"identity":"18c270b7-ff16-4a75-bd6d-977a77a6e8ce","order_by":9,"name":"Carleen Sabusap","email":"","orcid":"","institution":"University of Vanderbilt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Carleen","middleName":"","lastName":"Sabusap","suffix":""},{"id":2853082,"identity":"8c3303e5-fd23-420a-8c25-25965f5665a0","order_by":10,"name":"Francisca Bermedo","email":"","orcid":"","institution":"University of Concepcion","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Francisca","middleName":"","lastName":"Bermedo","suffix":""},{"id":2853083,"identity":"b1c41ce0-cb9a-4b84-924d-8eff57139a08","order_by":11,"name":"Stuart Adamson","email":"","orcid":"","institution":"Buck Institute for Research on Aging","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Stuart","middleName":"","lastName":"Adamson","suffix":""},{"id":2853084,"identity":"01fdb175-cb82-4984-aacf-8a6f2715f7d1","order_by":12,"name":"Kaitlyn Vitangcol","email":"","orcid":"","institution":"Buck Institute for Research on Aging","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Kaitlyn","middleName":"","lastName":"Vitangcol","suffix":""},{"id":2853085,"identity":"ac1bd82f-217e-4fe3-a561-5bf773ba0f8f","order_by":13,"name":"Hernan Huerta","email":"","orcid":"","institution":"University Mayor","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hernan","middleName":"","lastName":"Huerta","suffix":""},{"id":2853086,"identity":"114ee8ca-8d03-45fc-8e44-feefb5c24fb8","order_by":14,"name":"Xu Zhang","email":"","orcid":"","institution":"The Scripps Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xu","middleName":"","lastName":"Zhang","suffix":""},{"id":2853087,"identity":"78ea5577-0402-4933-953b-c41caa96969f","order_by":15,"name":"Tomohiro Nakamura","email":"","orcid":"","institution":"The Scripps Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tomohiro","middleName":"","lastName":"Nakamura","suffix":""},{"id":2853088,"identity":"fab40f01-0bb1-4ca7-8647-3fd8c4d35daf","order_by":16,"name":"Pablo Sardi","email":"","orcid":"","institution":"Rare and Neurological Diseases Therapeutic Area, Sanofi,","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Pablo","middleName":"","lastName":"Sardi","suffix":""},{"id":2853089,"identity":"4f170b09-be14-4ff4-99a2-8311dd912973","order_by":17,"name":"Stuart Lipton","email":"","orcid":"","institution":"The Scripps Research Institute","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Stuart","middleName":"","lastName":"Lipton","suffix":""},{"id":2853090,"identity":"f621c2e3-90d6-47ca-ad8e-67765dd4bdf5","order_by":18,"name":"Brian Kennedy","email":"","orcid":"","institution":"Buck Institute for Research on Aging","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Brian","middleName":"","lastName":"Kennedy","suffix":""},{"id":2853091,"identity":"c51fa92a-1d51-460c-ac0d-bb8ae3da8b47","order_by":19,"name":"Julio Matus","email":"","orcid":"","institution":"University Mayot","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Julio","middleName":"","lastName":"Matus","suffix":""},{"id":2853092,"identity":"a8660ea2-650b-4f2d-a987-0788ff81c841","order_by":20,"name":"Adrian Palacios","email":"","orcid":"","institution":"University of Valparaiso","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Adrian","middleName":"","lastName":"Palacios","suffix":""},{"id":2853093,"identity":"523da165-0072-4a30-93a4-905cafc6afba","order_by":21,"name":"Lars Plate","email":"","orcid":"","institution":"University of Vanderbilt","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Lars","middleName":"","lastName":"Plate","suffix":""},{"id":2853094,"identity":"1f7e094b-250a-4036-9eca-4130a3e2a135","order_by":22,"name":"Juan Pablo Henriquez","email":"","orcid":"","institution":"University of Concepcion","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Juan","middleName":"Pablo","lastName":"Henriquez","suffix":""},{"id":2853095,"identity":"07018da6-61c2-4352-89ba-ea028da725f0","order_by":23,"name":"Claudio Hetz","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAt0lEQVRIiWNgGAWjYDACCRBRwcBgwMzcAGRZEKWFsYHhDEgLYwPMBCK0MLYBtTAQq4V/dvPxhz/nHZY3Z2dsfMBQQ4QWiTvHEhsktx023NnM2GzAcIwILQYSOYYNhtsOJxgcZmyTYGwgSkv+x4bEOWAt7T+I1JLD2HCwAWILA1FaJG6kGc5sOJZuuOEwY7NEAjF+4Z+R/ODjjxpreYPzhw9++FBjQ1gLKkggVcMoGAWjYBSMAuwAAIVROFCzMrTRAAAAAElFTkSuQmCC","orcid":"","institution":"University of Chile","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Claudio","middleName":"","lastName":"Hetz","suffix":""}],"badges":[],"createdAt":"2020-09-25 15:18:00","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-83658/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-83658/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":2833292,"identity":"044e7e48-2920-423f-9b1e-3f3014d78138","added_by":"auto","created_at":"2020-10-07 16:32:57","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":108719,"visible":true,"origin":"","legend":"Impaired UPR activation in the mammalian aged brain. (a) Functional enrichment analysis of altered genes in an elderly cohort of patients diagnosed with dementia versus non-demented. GO: gene ontology; Key TF: key transcription factor in TRRUST database; FDR: false discovery rate. (b) Violin plots displaying normalized FPKM values of chaperones HSPA1a and HSPA1b in RNA-sequencing of human hippocampus of demented patients and controls (n = 49,42; unpaired Kolmogorov-Smirnov test, **: P \u003c0.01). (c-e) Young, middle-aged, and aged animals were treated with tunicamcyin (5 mg/kg) or vehicle. After 24 h, the mRNA levels of Xbp1-s, Bip/Hspa5, Chop/Ddit3 and Atf3 were determined in dissected hippocampus by quantitative RT-PCR. (n = 4 animals/group. One-way ANOVA followed by Tukey’s post-test compared tunicamycin treated groups, **: P \u003c 0.01). (f, g) Relative levels of S-nitrosylated IRE1α (d) and total IRE1α (e) in brains from young and aged mice. Histograms show relative levels of SNO-IRE1α/input IRE1α measured by biotin-switch assay (f) and total IRE1α/GAPDH quantified by densitometry on standard immunoblots (g) for young (3 month-old) versus aged (24 month-old) mouse brains (n = 3, *: P \u003c 0.05 by Student’s t test). ","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1/e39136b293272f56d5b77b10.png"},{"id":2833293,"identity":"b65cb644-1094-4cfe-80a2-deb6205832ef","added_by":"auto","created_at":"2020-10-07 16:32:57","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":156444,"visible":true,"origin":"","legend":"Genetic ablation of the IRE1α pathway in the brain accelerates and exacerbates age-associated cognitive and motor decline in mammals. (a) Conditional knockout mice for IRE1α were generated using the Nestin-CRE system (IRE1cKO) and their littermate flox/flox control animals (IRE1WT). The new object recognition test was used to evaluate the ability to discriminate novel objects after 24 h in young, middle-aged and aged mice (n = 10/ group; ***: P \u003c 0.005 by two-way ANOVA followed by Sidak’s multiple comparison test). (b) In animals of indicated genotypes, contextual fear conditioning test was performed to compare total number of freezing episodes following 24 h of aversive stimuli in young (n = 7, 10) and middle-aged (n = 9, 14) wild-type (IRE1WT) or IRE1cKO mice (*P \u003c 0.05 by unpaired Student’s t test within each age-matched group). (c) Young (n = 9, 9), middle-aged (n = 9, 9), and aged (n = 10, 8) IRE1WT or IRE1cKO mice were evaluated in the Barnes maze to evaluate spatial memory acquisition. Total number of errors before finding the target hole is plotted (*P \u003c 0.05 by unpaired Student’s t test within each age-matched group). (d) Young WT (n = 8), young XBP1scKO (n = 12), middle-aged WT (n = 18), or middle-aged XBP1cKO (n = 14) mice were evaluated in the new object recognition test to analyze the ability to discriminate between novel objects following 24 h of presentation of two identical objects (*: P \u003c 0.05 by unpaired Student’s t test comparing age-matched groups). (e) Scheme to illustrate the loss-of-function approach based on AAV2-CRE-mediated deletion of floxed IRE1α in the hippocampus. IRE1flox/flox animals were evaluated using the NOR test and then received bilateral hippocampal injections with AAV2-CRE. After 4 weeks, animals were evaluated again in the NOR test and then euthanized. Brains were collected for dendritic spine and biochemical analysis. (f) Young (n = 7) and middle-aged (n = 6) IRE1flox/flox animals were evaluated in the NOR test before and after AAV2-CRE intra-hippocampal injections (*: P \u003c 0.05 by paired Student’s t test comparing each individual before and after AAV2 injection). (g) Animals of different ages were injected with adeno-associated vector (AAV-CRE or AAV2-Mock) into the CA1 region of each hippocampus. AAV constructs included a GFP cassette to express eGFP in neurons for monitoring dendritic spine density. One month after the injection, spines were imaged by confocal microscopy. Left-hand panels: Representative confocal microscopic images of dendritic spines (arrows) in the CA1 region of middle-aged animals. 60x magnification; scale bar, 5 µm. Right-hand panel: Histogram of mean and SEM of spine density per µm (n = 24 dendrites, 4 animals; 27 dendrites, 3 animals; 31 dendrites, 4 animals; 42 dendrites, 4 animals, respectively). ****: P \u003c 0.001 by unpaired Student’s t test within each age-matched group. (h) Quantitative PCR showing Bdnf relative mRNA levels comparing young, middle aged or aged IRE1cKO to controls (n = 3-4 animals/group. Unpaired Student’s t test was performed comparing age-matched groups, *: P \u003c 0.05). (i) Wire hanging test performed in young, middle-aged, and aged IRE1WT or IRE1cKO to evaluate motor performance. Graph indicates mean and SEM of arbitrary scores from aged (n = 18, 15), middle-aged (n = 11, 15), and young (n = 10, 12) animals (*: P \u003c 0.05 by two-way ANOVA followed by Sidak’s post-hoc test). (j) Rotarod test used to evaluate motor performance and coordination in young (n = 16, 12), middle-aged (n = 8, 8), and aged (n = 18, 6) IRE1WT or IRE1cKO animals. Graph indicates mean and SEM of latencies to fall from the rod (*: P \u003c 0.05 by unpaired Student’s t test within each age group). (k) Representative photomicrographs of β-galactosidase staining of hippocampal slices derived from young and middle-aged IRE1WT or IRE1cKO animals (n = 3-4 animals/group). Graphs (at right) indicate mean and SEM of percentage of β-galactosidase positive cells. **: P \u003c 0.01 by unpaired Student’s t test within each age-matched group for each hippocampal sub-region. Magnification: 20x; scale bar: 200 µm.","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1/13be2a394771935a4840ac17.png"},{"id":2833294,"identity":"ed9509e1-a720-4d55-985f-4e99df9fb6f3","added_by":"auto","created_at":"2020-10-07 16:32:57","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":86867,"visible":true,"origin":"","legend":"Enforced expression of XBP1s in the brain prevents age-associated motor and cognitive decline in mammals. (a) New object recognition test was used to evaluate ability to discriminate novel objects after 24 h in TgXBP1s or littermate non-transgenic animals (non-Tg) in young, middle-aged, and aged animals (n = 22, 6; 23, 9; 17, 12; respectively; *: P \u003c 0.05 by unpaired Student’s t test within each age-matched group). (b) New object location test was used to evaluate the ability to discriminate changes in object location after 24 h in young, middle-aged, and aged non-Tg or TgXBP1s (n = 20, 7; 12, 13; 19, 9 respectively; histogram shows mean and SEM of percentage of time interacting with the novel-located object (NLO). *: P \u003c 0.05 by unpaired Student’s t test within each age-matched group). (c) Aged non-Tg and aged TgXBP1s animals were evaluated in the Barnes maze test to compare spatial memory acquisition over a period of 5 days. Graph indicates mean and SEM of each group (n = 10 for non-Tg [WT], 8 for TgXBP1s). Student’s t test was used to compare performances within each day (*P \u003c 0.05). Latencies of young/non-Tg mice also plotted for comparison (n = 10). (d) Young, middle-aged, and aged non-Tg or TgXBP1s were evaluated in the wire hanging test to monitor motor performance and coordination. Mean and SEM of arbitrary scores for each group (n = 8 animals/group; *: P \u003c 0.05, **P \u003c 0.01 by two-way ANOVA followed by Sidak’s multiple comparison test). (e) Young and aged non-Tg or TgXBP1s animals were evaluated in the rotarod test to monitor motor performance and coordination. Mean and SEM for each group (n = 10 animals/group, ***: P \u003c 0.001 by unpaired Student’s t test within each matched group). (f) Following MS/MS analysis of the hippocampal proteome, gene set enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes (KEGG) library. Graphs depict most enriched terms based on combined scored computed by EnrichR platform comparing middle aged and aged TgXBP1s with age-matched controls. Terms were divided in two graphs related to synaptic function or neurodegeneration. (g) Protein-protein interaction networks depicting main genes altered in middle aged and aged TgXBP1s (PPI enrichment p-value \u003c 1-16). ","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1/955caf72a6fe370ee2d2b670.png"},{"id":2833295,"identity":"f36aa9af-3daf-4f34-a4ab-edfae5180370","added_by":"auto","created_at":"2020-10-07 16:32:58","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":206689,"visible":true,"origin":"","legend":"XBP1s-gene delivery to aged animals reverses age-associated phenotypes at the behavioral,\nmorphological and electrophysiological levels. (a) Middle-aged and aged WT animals were injected with AAV2-\nMock or AAV2-XBP1s in both hippocampi and then evaluated in the new object location test to evaluate their\nability to discriminate displacement of objects 24 h following presentation. Histogram shows mean and SEM of\npercentage of time interacting with the novel located object (NLO) for n = 10 animals/group, **: P \u003c 0.01 by\nunpaired Student’s t test). Young animal performance plotted for comparison (n = 6). (b) Aged animals injected\nwith AAV2-Mock or AAV2-XBP1s in both hippocampi were evaluated in the new object recognition test to evaluate their ability to discriminate novel objects after 24 h. Mean and SEM of percentage of interaction time with novel objects (n = 10 animals/group, ***: P \u003c 0.005 by unpaired Student’s t test). Young animal performance plotted for comparison (n = 6). (c) Aged animals injected with AAV2-Mock or AAV2-XBP1s in both hippocampi were evaluated in the Barnes maze test to compare spatial memory acquisition. Mean and SEM of latencies to find target for each group during 4 days of testing (n = 5 for AAV2-Mock, 8 for AAV2-XBP1s; Student’s t test was used to compare performances within each day (*: P \u003c 0.05). Young animal performance plotted for comparison (n = 10). (d) Aged animals injected with AAV2-Mock or AAV2-XBP1s evaluated by contextual fear conditioning for aversive memory acquisition 24 h after presentation of an unconditioned stimulus (n = 5 for AAV2-Mock, 7 for AAV2-XBP1s; **: P \u003c 0.01 by unpaired Student’s t test). Young animal performance plotted for comparison (n = 6). (e) Brain slice electrophysiological analysis assessing firing rates in hippocampal neurons. Mean and SEM of firing rates were measured during spontaneous activity or following picrotoxin treatment in brain slices from aged mice injected with AAV2-Mock or AAV2-XBP1s (n = 633-832 neurons; n = 6, 9 animals, respectively; ****: P \u003c 0.001 by unpaired Student’s t test). (f) Basal and picrotoxin-induced burst activity measured in pyramidal and interneurons in hippocampal brain slices derived from aged mice injected with AAV2-Mock or AAV2-XBP1s (n = 832, 644 neurons from 3-4 animals, **: P \u003c 0.01 by unpaired Student’s t test). (g) Mean and SEM of field excitatory postsynaptic potentials amplitudes in brain slices derived from aged animals injected with AAV2-Mock or AAV2-XBP1s 1 h after theta burst stimulus (TBS) to induce LTP in (n = 6, 9 animals, respectively; n = 17-28 slices/animal, ****: P \u003c 0.001 by unpaired Student’s t test). (h) Representative fluorescent images showing dendritic spines of pyramidal neurons in CA1 region of young and aged animals injected with AAV2-eGFP (mock) or aged animals injected with AAV2-XBP1s. Young animal neuron is shown for reference. Right panel: Mean and SEM of spine density per µm for indicated experimental groups (n = 24, 53 and 35 dendrites from 3, 6 and 5 animals, respectively; **: P \u003c 0.01 by unpaired Student’s t test comparing the aged groups). 60x magnification; scale bar, 5 µm). (i) Representative images of β-galactosidase staining of hippocampal slices derived from aged mice injected with AAV2 (n = 3-4 animals/group). Histograms (right) show mean and SEM of percentage of β-galactosidase positive cells. Magnification 20x; scale bar, 200 µm. (j) Following MS/MS analysis of the hippocampal proteome, gene set enrichment analysis was performed using Kyoto Encyclopedia of Genes and Genomes (KEGG) library. Graphs depict most enriched terms based on combined scored computed by EnrichR platform comparing aged mice infected with AAV-Mock or AAV-XBP1s. Terms were divided in two graphs related to synaptic function or neurodegeneration. (k) Volcano plot for proteomic comparison of hippocampal tissue derived from aged animals injected with AAV-Mock or AAV-XBP1s. FDR (y-axis) and fold-change (log2) (x-axis) are indicated. Dotted lines delineate cut-off used to filter genes for functional enrichment analysis. Indicated genes associated with each enriched terms connected to synaptic function, neurodegeneration, metabolism and ECM-receptor interaction are colored.","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1/fd17a4bd31105061d0a54386.png"},{"id":13539341,"identity":"e04e2502-4a3b-4742-bf31-778ce7c8dcbf","added_by":"auto","created_at":"2021-09-17 01:43:03","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8567967,"visible":true,"origin":"","legend":"","description":"","filename":"CabralMirandaetal2020NatureAgingmaintexandfigureswithsupplelmentarymaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1_covered.pdf"},{"id":11965361,"identity":"a9eb3119-e60d-4fb5-b560-eb519222e670","added_by":"auto","created_at":"2021-07-30 17:55:59","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":8564380,"visible":true,"origin":"","legend":"","description":"","filename":"CabralMirandaetal2020NatureAgingmaintexandfigureswithsupplelmentarymaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1_covered.pdf"},{"id":2833299,"identity":"5b5ec389-3fed-41de-81ba-d5ab298ff04c","added_by":"auto","created_at":"2020-10-07 16:33:03","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":15257465,"visible":true,"origin":"","legend":"","description":"","filename":"CabralMirandaetal2020NatureAgingmaintexandfigureswithsupplelmentarymaterial.pdf","url":"https://assets-eu.researchsquare.com/files/rs-83658/v1_stamped.pdf"}],"financialInterests":"\u003cb\u003eYes\u003c/b\u003e there is potential Competing Interest.\nCH and FCM declare a conflict of interest for a submitted patent application. Inventors: Claudio Hetz, Felipe Cabral Miranda. Title: Treatment of aging or age- related disorders using XBP1. Provisional application for patent at USPTO, application number 62800229. Submitted 01/02/2019. Status: patent pending.\r\nDisclosure: PS is a Sanofi employee.","formattedTitle":"Control of mammalian brain ageing by the unfolded protein response transcription factor XBP1","fulltext":[{"header":"Full Text","content":"\u003cp\u003eThis preprint is available for \u003ca href='/article/rs-83658/latest.pdf' target='_blank'\u003edownload as a PDF\u003c/a\u003e.\u003c/p\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":true,"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":"Neurodegenerative Disease, Proteostatsis Network, ER Stress","lastPublishedDoi":"10.21203/rs.3.rs-83658/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-83658/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"Brain ageing is the main risk factor to develop dementia and neurodegenerative diseases, associated with a decay in the buffering capacity of the proteostasis network. We investigated the significance of the unfolded protein response (UPR), a major signaling pathway to cope with ER stress, to the functional deterioration of the brain during aging. Genetic disruption of the ER stress sensor IRE1α accelerated cognitive and motor decline during ageing. Exogenous bolstering of the UPR by overexpressing an active form of the UPR transcription factor XBP1 restored synaptic and cognitive function, in addition to reducing cell senescence. Proteomic profiling of hippocampal tissue indicated that XBP1s expression attenuated age-related alterations to synaptic function and pathways linked to neurodegenerative diseases. Overall, our results demonstrate that strategies to manipulate the UPR in mammals may sustain healthy brain\r\nageing.","manuscriptTitle":"Control of mammalian brain ageing by the unfolded protein response transcription factor XBP1","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2020-10-07 16:32:56","doi":"10.21203/rs.3.rs-83658/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":"abe00738-172b-40b4-9965-067c3216a5af","owner":[],"postedDate":"October 7th, 2020","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":680678,"name":"Cognitive Neuroscience"},{"id":680679,"name":"Cellular \u0026 Molecular Neuroscience"}],"tags":[],"updatedAt":"2022-01-27T17:55:45+00:00","versionOfRecord":[],"versionCreatedAt":"2020-10-07 16:32:56","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-83658","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-83658","identity":"rs-83658","version":["v1"]},"buildId":"FbvkV6FR0MCFSLy54lSbu","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}