Cholesterol Modulates Galanin Receptor Subtype 1 but Not Subtype 2: Insights from Live-Cell Imaging

Cholesterol Modulates Galanin Receptor Subtype 1 but Not Subtype 2: Insights from Live-Cell Imaging | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL British Journal of Pharmacology This is a preprint and has not been peer reviewed. Data may be preliminary. 26 February 2025 V1 Latest version Share on Cholesterol Modulates Galanin Receptor Subtype 1 but Not Subtype 2: Insights from Live-Cell Imaging Authors : Tianyi Li [email protected] , Vladana Vukojević , Sho Oasa , Yunfei Bai , Yutao Yang , Hui Li , Yi Tang , Lars Terenius , Tomas Hökfelt , Per Svenningsson , and Zhiqing Xu Authors Info & Affiliations https://doi.org/10.22541/au.174060368.80090212/v1 Published British Journal of Pharmacology Version of record Peer review timeline 350 views 197 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract Abstract Background and Purpose Galanin receptor subtypes 1 (GALR1) and 2 (GALR2) are G protein-coupled receptors (GPCRs) that mediate galanin’s diverse physiological roles, including neurotransmission and neuronal modulation. Although both receptors share functional similarities, they exhibit distinct differences in signaling pathways. While previous studies have focused on galanin binding and G-protein selectivity, the role of plasma membrane-specific mechanisms, particularly cholesterol’s influence, remains unclear. This study investigates cholesterol’s role in regulating GALR1 and GALR2 trafficking and function in live cells. Experimental Approach We employed real-time fluorescence techniques—Fluorescence Correlation Spectroscopy (FCS), Fluorescence Cross-Correlation Spectroscopy (FCCS), and Fluorescence Recovery After Photobleaching (FRAP)—to assess receptor-ligand interactions and lateral mobility in PC12 cells expressing EGFP-tagged GALR1 or GALR2. Key Results Both receptors co-localized, co-trafficked, and internalized with galanin, with receptor-peptide complexes dissociating prior to lysosomal degradation. Cholesterol selectively restricted GALR1’s lateral diffusion and enhanced galanin binding and complex formation, while GALR2 remained unaffected. Interestingly, galanin binding relieved GALR1 from cholesterol-mediated restriction, increasing receptor mobility and suggesting a dynamic, cholesterol-dependent regulatory mechanism. Conclusions and Implications Cholesterol selectively modulates GALR1 trafficking and ligand interactions, while GALR2 operates independently of cholesterol, revealing distinct regulatory mechanisms for each receptor subtype. These findings provide new insights into the interplay between membrane composition and receptor function, with potential implications for developing targeted therapies for galanin-related disorders. Supplementary Material File (main text and figures.docx) Download 4.52 MB Information & Authors Information Version history V1 Version 1 26 February 2025 Peer review timeline Published British Journal of Pharmacology Version of Record 18 Sep 2025 Published Copyright This work is licensed under a Non Exclusive No Reuse License. Collection British Journal of Pharmacology Keywords depression gpcr imaging ligands neuropharmacology receptor theory receptor/channel trafficking Authors Affiliations Tianyi Li [email protected] Capital Medical University View all articles by this author Vladana Vukojević Karolinska Institute View all articles by this author Sho Oasa Karolinska Institute View all articles by this author Yunfei Bai Capital Medical University View all articles by this author Yutao Yang Capital Medical University View all articles by this author Hui Li Capital Medical University View all articles by this author Yi Tang Xuanwu Hospital Capital Medical University View all articles by this author Lars Terenius Karolinska Institute View all articles by this author Tomas Hökfelt Karolinska Institute View all articles by this author Per Svenningsson Karolinska University Hospital View all articles by this author Zhiqing Xu Capital Medical University View all articles by this author Metrics & Citations Metrics Article Usage 350 views 197 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Tianyi Li, Vladana Vukojević, Sho Oasa, et al. Cholesterol Modulates Galanin Receptor Subtype 1 but Not Subtype 2: Insights from Live-Cell Imaging. Authorea . 26 February 2025. DOI: https://doi.org/10.22541/au.174060368.80090212/v1 If you have the appropriate software installed, you can download article citation data to the citation manager of your choice. Simply select your manager software from the list below and click Download. For more information or tips please see 'Downloading to a citation manager' in the Help menu . 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