{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,3,12]],"date-time":"2026-03-12T15:44:21Z","timestamp":1773330261312,"version":"3.50.1"},"posted":{"date-parts":[[2026,3,9]]},"group-title":"Neuroscience","reference-count":113,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T00:00:00Z","timestamp":1773014400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH","award":["F31HL165896"],"award-info":[{"award-number":["F31HL165896"]}]},{"name":"NIH","award":["R01MH112739"],"award-info":[{"award-number":["R01MH112739"]}]},{"name":"NIH","award":["R01NS131664"],"award-info":[{"award-number":["R01NS131664"]}]},{"name":"NIH","award":["R03DC019290"],"award-info":[{"award-number":["R03DC019290"]}]},{"name":"NIH","award":["R01EY026817"],"award-info":[{"award-number":["R01EY026817"]}]},{"name":"NIH","award":["R03TR005086"],"award-info":[{"award-number":["R03TR005086"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,3,9]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                <jats:p>\n                  Synapse formation and function are coordinated spatially and temporally by a host of synaptic proteins that regulate neuronal signaling, synapse specificity, and plasticity; many of which are implicated in neuropsychiatric disorders. Members of the C1q\/TNF superfamily function as synaptic organizers, shaping synapse assembly and maintenance. Among them, C1QL3 plays a putative role in trans-synaptic adhesion and modulation of synaptic strength, but the lack of a reliable antibody to detect it has severely limited the ability to map its endogenous localization and study its biochemical properties. Here, we present a novel epitope-tagged knock-in mouse line (\n                  <jats:italic>C1ql3<\/jats:italic>\n                  <jats:sup>2HA<\/jats:sup>\n                  ), in which two hemagglutinin (HA) epitopes were inserted near the N-terminus of the endogenous C1QL3 protein. This model enables purification, detection, and subcellular localization of native C1QL3 protein (C1QL3-2HA) with high specificity, eliminating the need for overexpression or custom antibodies. We validated that\n                  <jats:italic>C1ql3<\/jats:italic>\n                  <jats:sup>2HA<\/jats:sup>\n                  mice maintain normal mRNA expression, biochemical properties, and behavior. Using native PAGE, we determined the endogenous oligomeric state of C1QL3-2HA. Brain-wide light-sheet microscopy uncovered an expanded neuroanatomical map of C1QL3-2HA expression, including newly identified populations in cortical and subcortical regions as well as the retina. Dual immunohistochemistry confirmed cell type-specific expression patterns, and super-resolution STED microscopy localized C1QL3-2HA to hippocampal mossy fiber synapses, positioned between pre- and post-synaptic markers, supporting its hypothesized role in trans-synaptic complexes. This knock-in mouse line is a powerful tool for studying the anatomical, molecular, and synaptic biology of C1QL3 in all cellular\/tissue contexts, enabling future studies into its potential roles in the nervous system and beyond.\n                <\/jats:p>","DOI":"10.64898\/2026.03.05.709958","type":"posted-content","created":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T20:55:13Z","timestamp":1773089713000},"source":"Crossref","is-referenced-by-count":0,"title":["Brain-wide mapping and synaptic localization of C1QL3 using a novel epitope-tagged knock-in mouse"],"prefix":"10.64898","author":[{"suffix":"IV","given":"William P.","family":"Armstrong","sequence":"first","affiliation":[{"name":"Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT 06269, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"James M.","family":"Salvatore","sequence":"additional","affiliation":[{"name":"Department of Physiology and Neurobiology, University of Connecticut, Storrs, CT 06269, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Matthew J.","family":"Sticco","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Connecticut Health, Farmington, CT 06030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Keaven","family":"Caro","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Connecticut Health, Farmington, CT 06030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. Wesley","family":"Maddox","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas-Austin, Austin, TX 78712, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Angie","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas-Austin, Austin, TX 78712, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brenna C.","family":"McAllister","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Connecticut Health, Farmington, CT 06030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Christopher","family":"O\u2019Connell","sequence":"additional","affiliation":[{"name":"Advanced Light Microscopy Facility, Center for Open Research Resources and Equipment, University of Connecticut, Storrs, CT 06269, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Siu-Pok","family":"Yee","sequence":"additional","affiliation":[{"name":"Center for Mouse Genome Modification, UConn Health, Farmington, CT 06030, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8021-0443","authenticated-orcid":false,"given":"Amy","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas-Austin, Austin, TX 78712, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Susanne","family":"Ressl","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Texas-Austin, Austin, TX 78712, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"David C.","family":"Martinelli","sequence":"additional","affiliation":[{"name":"Department of Neuroscience, University of Connecticut Health, Farmington, CT 06030, USA"},{"name":"The Connecticut Institute for 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