{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,2,18]],"date-time":"2026-02-18T10:13:46Z","timestamp":1771409626100,"version":"3.50.1"},"posted":{"date-parts":[[2026,2,16]]},"group-title":"Neuroscience","reference-count":45,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T00:00:00Z","timestamp":1771200000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100000065","name":"National Institute of Neurological Disorders and Stroke","doi-asserted-by":"crossref","award":["NS077114"],"award-info":[{"award-number":["NS077114"]}],"id":[{"id":"10.13039\/100000065","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,2,16]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>\n                  The serotonin type 3A (5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  ) receptor is a pentameric ligand-gated ion channel (pLGIC) in central and peripheral neurons that conducts sodium and potassium ions upon serotonin binding. 5-HT3 receptors modulate neurotransmission and synaptic plasticity, influencing mood, sleep, appetite, and addiction. Disruptions in serotonin signaling are linked to central nervous system disorders, including schizophrenia, anxiety and depression. Clinically, these receptors are targeted by antagonists to treat chemotherapy-induced nausea and vomiting. The functional surface expression of these channels is regulated by the chaperone protein Resistant to Inhibitors of Cholinesterase 3 (RIC-3) that promotes plasma-membrane expression, maturation, and trafficking of 5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  and nicotinic acetylcholine receptors. Our previous work identified a duplicated RIC-3 binding motif within the 5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  intracellular domain (ICD). However, it was unclear whether this interaction reflected native conditions. Here, we used a recombinant 5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  ICD peptide in peptide-resin pull-down assays to investigate RIC-3 Interactions in plasma membrane (PM) fractions from\n                  <jats:italic>Xenopus<\/jats:italic>\n                  oocytes, endoplasmic reticulum (ER) fractions from SH-SY5Y cells, and mouse brain tissue. Across all tested systems, the 5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  ICD peptide specifically bound RIC-3. Furthermore, RIC-3 knockdown (RIC-3 KD) SH-SY5Y cells showed a marked reduction in peptide binding and decreased surface levels of nAChR\u03b17 and 5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  receptors. These results demonstrate RIC-3-5-HT\n                  <jats:sub>3A<\/jats:sub>\n                  ICD interaction in native cellular contexts and support a role for RIC-3 in regulating receptor surface expression and neuronal signaling.\n                <\/jats:p>","DOI":"10.64898\/2026.02.15.706004","type":"posted-content","created":{"date-parts":[[2026,2,16]],"date-time":"2026-02-16T10:00:13Z","timestamp":1771236013000},"source":"Crossref","is-referenced-by-count":0,"title":["RIC-3 Interacts Directly with the 5-HT\n                  <sub>3A<\/sub>\n                  Receptor to Mediate Trafficking Across Subcellular Compartments"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0005-4028-2705","authenticated-orcid":false,"given":"Nermina","family":"Jahovic","sequence":"first","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3898-9574","authenticated-orcid":false,"given":"Petar N.","family":"Grozdanov","sequence":"additional","affiliation":[{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Department of Pharmacology and Neuroscience, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Rhea","family":"Ramani","sequence":"additional","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Cade","family":"Perkins","sequence":"additional","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3209-1982","authenticated-orcid":false,"given":"Clinton C.","family":"MacDonald","sequence":"additional","affiliation":[{"name":"Department of Cell Biology and Biochemistry; Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joshua","family":"Theriot","sequence":"additional","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7361-6066","authenticated-orcid":false,"given":"Quynh Hoa","family":"Do","sequence":"additional","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1325-9700","authenticated-orcid":false,"given":"Michaela","family":"Jansen","sequence":"additional","affiliation":[{"name":"Department of Cell Physiology and Molecular Biophysics, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"},{"name":"Center for Membrane Protein Research, Texas Tech University Health Sciences Center School of Medicine, Lubbock, TX, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"54368","reference":[{"key":"2026021801250912000_2026.02.15.706004v1.1","doi-asserted-by":"crossref","first-page":"4527","DOI":"10.3390\/molecules27144527","article-title":"Speculation on How RIC-3 and Other Chaperones Facilitate \u03b17 Nicotinic Receptor Folding and Assembly","volume":"27","year":"2022","journal-title":"Molecules"},{"key":"2026021801250912000_2026.02.15.706004v1.2","doi-asserted-by":"crossref","first-page":"2455","DOI":"10.3390\/biomedicines12112455","article-title":"Unraveling the Serotonergic Mechanism of Stress-Related Anxiety: Focus on Co-Treatment with Resveratrol and Selective Serotonin Reuptake Inhibitors","volume":"12","year":"2024","journal-title":"Biomedicines"},{"key":"2026021801250912000_2026.02.15.706004v1.3","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00015.2010"},{"key":"2026021801250912000_2026.02.15.706004v1.4","doi-asserted-by":"publisher","DOI":"10.1016\/S0074-7742(06)78008-4"},{"key":"2026021801250912000_2026.02.15.706004v1.5","doi-asserted-by":"publisher","DOI":"10.1038\/nature01788"},{"key":"2026021801250912000_2026.02.15.706004v1.6","doi-asserted-by":"crossref","unstructured":"Modular Design of Cys-loop Ligand-gated Ion Channels: Functional 5-HT 3 and GABA 1 Receptors Lacking the Large Cytoplasmic M3M4 Loop. 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