Behavioral regression in shank3Δex4-22 mice during early adulthood corresponds to cerebellar granule cell glutamatergic synaptic changes

Behavioral regression in shank3Δex4-22 mice during early adulthood corresponds to cerebellar granule cell glutamatergic synaptic changes | 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 Behavioral regression in shank3Δex4-22 mice during early adulthood corresponds to cerebellar granule cell glutamatergic synaptic changes Rajaram Kshetri, James Beavers, Romana Hyde, Roseline Ewa, Amber Schwertman, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4888950/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 04 Dec, 2024 Read the published version in Molecular Autism → Version 1 posted 10 You are reading this latest preprint version Abstract Background: Shank3, a gene encoding a synaptic scaffolding protein, is implicated in autism spectrum disorder (ASD) and is disrupted in Phelan-McDermid syndrome (PMS). Despite evidence of regression or worsening of ASD-like symptoms in individuals with PMS, the underlying mechanisms remain unclear. Although shank3 is highly expressed in the cerebellar cortical granule cells, its role in cerebellar function and contribution to behavioral deficits in ASD models are unknown. This study investigates behavioral changes and cerebellar synaptic alterations in shank3Δex4-22 mice at two developmental stages. Methods: Shank3Δex4-22 wildtype, heterozygous, and homozygous knockout mice lacking exons 4-22 (all functional isoforms) were subjected to a behavioral battery in both juvenile (5-7 weeks old) and adult (3-5 months old) mouse cohorts of both sexes. Immunostaining was used to show the expression of SHANK3 in the cerebellar cortex. Spontaneous excitatory postsynaptic currents (sEPSCs) from cerebellar granule cells (CGCs) were recorded by whole-cell patch-clamp electrophysiology. Results: Deletion of shank3ex4-22 caused deficits in motor function, heightened anxiety, and repetitive behaviors. These genotype-dependent behavioral alterations were more prominent in adult mice than in juveniles. Reduced social preference was only identified in adult shank3Δex4-22 knockout mice and self-grooming was uniquely elevated only in males across both age groups. Immunofluorescence staining indicates the presence of SHANK3 predominantly in the dendrite-containing rosette-like structures in CGCs, colocalizing with presynaptic markers of glutamatergic mossy fiber. Electrophysiological findings identify a parallel relationship between the age-related exacerbation of behavioral impairments and the enhancement of sEPSC amplitude in CGCs. Limitations: Other behavioral tests of muscle strength (grip strength test), memory (Barnes/water maze), and communication (ultrasonic vocalization), were not performed. Further study is necessary to elucidate how SHANK3 modulates synaptic function at the mossy fiber-granule cell synapse in the cerebellum. Conclusions: Our findings reveal an age-related exacerbation of behavioral impairments in shank3Δex4-22 mutant mice. These results suggest that SHANK3 may play a role in maintaining glutamatergic receptors and synapses in CGCs, as well as the potential involvement of the cerebellum in ASD. Shank3 autism spectrum disorder Phelan-McDermid syndrome regression cerebellum glutamate receptor AMPAR granule cell mouse behavior phenotype Full Text Additional Declarations No competing interests reported. Supplementary Files Shank3BehaviorStatsSummary.pdf Cite Share Download PDF Status: Published Journal Publication published 04 Dec, 2024 Read the published version in Molecular Autism → Version 1 posted Editorial decision: Revision requested 25 Sep, 2024 Reviews received at journal 25 Sep, 2024 Reviews received at journal 20 Sep, 2024 Reviews received at journal 12 Sep, 2024 Reviewers agreed at journal 28 Aug, 2024 Reviewers agreed at journal 23 Aug, 2024 Reviewers invited by journal 16 Aug, 2024 Editor assigned by journal 16 Aug, 2024 Submission checks completed at journal 12 Aug, 2024 First submitted to journal 09 Aug, 2024 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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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-4888950","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":348638325,"identity":"87f50a57-6307-4d8c-afe8-02ea8f718c99","order_by":0,"name":"Rajaram Kshetri","email":"","orcid":"","institution":"Southern Illinois University – School of Medicine","correspondingAuthor":false,"prefix":"","firstName":"Rajaram","middleName":"","lastName":"Kshetri","suffix":""},{"id":348638326,"identity":"0cdfa213-98b5-4041-8e67-20c174bbcc3d","order_by":1,"name":"James Beavers","email":"","orcid":"","institution":"Southern Illinois University – School 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Despite evidence of regression or worsening of ASD-like symptoms in individuals with PMS, the underlying mechanisms remain unclear. Although shank3 is highly expressed in the cerebellar cortical granule cells, its role in cerebellar function and contribution to behavioral deficits in ASD models are unknown. This study investigates behavioral changes and cerebellar synaptic alterations in shank3Δex4-22 mice at two developmental stages.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eMethods: Shank3Δex4-22 wildtype, heterozygous, and homozygous knockout mice lacking exons 4-22 (all functional isoforms) were subjected to a behavioral battery in both juvenile (5-7 weeks old) and adult (3-5 months old) mouse cohorts of both sexes. Immunostaining was used to show the expression of SHANK3 in the cerebellar cortex. Spontaneous excitatory postsynaptic currents (sEPSCs) from cerebellar granule cells (CGCs) were recorded by whole-cell patch-clamp electrophysiology.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eResults: Deletion of shank3ex4-22 caused deficits in motor function, heightened anxiety, and repetitive behaviors. These genotype-dependent behavioral alterations were more prominent in adult mice than in juveniles. Reduced social preference was only identified in adult shank3Δex4-22 knockout mice and self-grooming was uniquely elevated only in males across both age groups. Immunofluorescence staining indicates the presence of SHANK3 predominantly in the dendrite-containing rosette-like structures in CGCs, colocalizing with presynaptic markers of glutamatergic mossy fiber. 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These results suggest that SHANK3 may play a role in maintaining glutamatergic receptors and synapses in CGCs, as well as the potential involvement of the cerebellum in ASD.\u003c/p\u003e","manuscriptTitle":"Behavioral regression in shank3Δex4-22 mice during early adulthood corresponds to cerebellar granule cell glutamatergic synaptic changes","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-09-06 19:24:09","doi":"10.21203/rs.3.rs-4888950/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2024-09-25T15:23:27+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-25T15:04:13+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-20T09:01:32+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2024-09-12T16:35:37+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"110609995594657147600888146673865611028","date":"2024-08-28T13:31:00+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"299523067898609059256224241041991176225","date":"2024-08-23T13:02:10+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2024-08-16T16:19:14+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2024-08-16T16:16:10+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2024-08-12T06:52:32+00:00","index":"","fulltext":""},{"type":"submitted","content":"Molecular Autism","date":"2024-08-09T19:57:36+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"molecular-autism","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"mola","sideBox":"Learn more about [Molecular Autism](http://molecularautism.biomedcentral.com/)","snPcode":"13229","submissionUrl":"https://submission.nature.com/new-submission/13229/3","title":"Molecular Autism","twitterHandle":"@MolecularAutism","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"BMC/SO AJ","inReviewEnabled":true,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"850af63f-2ca7-4e4a-a45c-c7eb1ac1bef8","owner":[],"postedDate":"September 6th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-12-09T16:06:28+00:00","versionOfRecord":{"articleIdentity":"rs-4888950","link":"https://doi.org/10.1186/s13229-024-00628-y","journal":{"identity":"molecular-autism","isVorOnly":false,"title":"Molecular Autism"},"publishedOn":"2024-12-04 15:58:05","publishedOnDateReadable":"December 4th, 2024"},"versionCreatedAt":"2024-09-06 19:24:09","video":"","vorDoi":"10.1186/s13229-024-00628-y","vorDoiUrl":"https://doi.org/10.1186/s13229-024-00628-y","workflowStages":[]},"version":"v1","identity":"rs-4888950","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4888950","identity":"rs-4888950","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}