Single-molecule detection of transient dimerization of opioid receptors 2: Heterodimer blockage reduces morphine tolerance

Single-molecule detection of transient dimerization of opioid receptors 2: Heterodimer blockage reduces morphine tolerance | 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 Single-molecule detection of transient dimerization of opioid receptors 2: Heterodimer blockage reduces morphine tolerance Peng Zhou, Rinshi S. Kasai, Wakako Fujita, Taka A. Tsunoyama, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4991895/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 Heterodimerization of opioid receptors (ORs), MOR, KOR, and DOR, is implied in their functional regulation and diversification, and thus its understanding is crucial for developing better analgesic treatments. However, our knowledge on OR heterodimerization/heterodimers remains limited. Here, using single-molecule imaging and functional analysis, we found that MOR, the main morphine receptor, repeatedly forms transient (≈250 ms) heterodimers with DOR every 1-10 seconds, but not with KOR, whereas DOR and KOR also form transient heterodimers. We obtained all the heterodimer-monomer equilibrium constants and rate constants with/without agonists. We identified the critical heterodimer binding sites in the extracellular domains, in addition to the less-specific transmembrane domains, and developed soluble peptide blockers for MOR-DOR and DOR-KOR heterodimerization, using amino-acid sequences mimicking the extracellular binding sites. With these peptide blockers, we dissected the monomer/dimer roles in OR internalization and signaling. The soluble MOR-DOR heterodimer blocker reduced the development of long-term morphine tolerance in mice. Biophysics Opioids Opioid receptor Single molecule imaging Heterodimer Heterodimer-monomer equilibrium constant Heterodimer blocker Tolerance Full Text Additional Declarations The authors declare potential competing interests as follows: P.Z. and A.K. have a patent pending for DpepDMs, MpepMDs and peptides from EL3 domains of ORs. The remaining authors have no conflicts of interest to declare. Supplementary Files heteroSupplementaryinformation.docx.pdf 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-4991895","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":346389331,"identity":"b4d0878e-1bff-427d-828e-063b57ec7283","order_by":0,"name":"Peng Zhou","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAzUlEQVRIie3RsQqCQBjA8Q+EbjmabfEZPjjQBOlZLgQnH8IHsHfpEU6+oUWaFYOCIBoabDNw6IyGtru2oPsPn3D4Az8PwOX63RQAK6YJ3heEKz3lV8SX+ikt3o3Yhq4wHgJsz1X1GIBFJhKX+2wJ/CKwyyRx/WFxYSDY5CGCT+ttlyNNu6AykeNNE9SkrbEarEjDxQmkJg1HxW1IXOYhSEViUU+7ZL55l4jVou9HCuY7ovuQJKnxjyHA7HUj7/wULYjXfx6sjMTlcrn+rieRz0P/T8R06wAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-3082-1462","institution":"Okinawa Institute of Science and Technology Graduate University","correspondingAuthor":true,"prefix":"","firstName":"Peng","middleName":"","lastName":"Zhou","suffix":""},{"id":346389332,"identity":"ac72d207-5ce5-420a-b4c2-7d293f90c18e","order_by":1,"name":"Rinshi S. 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