{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,2,21]],"date-time":"2025-02-21T02:26:32Z","timestamp":1740104792648,"version":"3.37.3"},"reference-count":25,"publisher":"Wiley","issue":"7","license":[{"start":{"date-parts":[[2019,7,13]],"date-time":"2019-07-13T00:00:00Z","timestamp":1562976000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Neurourology and Urodynamics"],"published-print":{"date-parts":[[2019,9]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Aims<\/jats:title><jats:p>To explore caveolae\u2010 and clathrin\u2010mediated internalization of muscarinic M2 and M3 receptors, recycling and degradation in formalin\u2010fixed paraffin\u2010embedded detrusor sections; to study alterations possibly involved in the pathophysiology of the bladder functional disorder, interstitial cystitis\/bladder pain syndrome (IC\/BPS).<\/jats:p><\/jats:sec><jats:sec><jats:title>Materials and Methods<\/jats:title><jats:p>Samples of IC\/BPS (n\u2009=\u200911) and cystectomy patients (n\u2009=\u200911) were analyzed. Proximity ligation assay (PLA) was used to detect interactions of M2 and M3 with endocytotic regulators (Cav\u20101, clathrin, Rab7, and Rab11) by Cy3 labeling. Analyses of three\u2010dimensional (3D)\u2010reconstructed z\u2010stacks (63\u2009\u00d7\u2009 Oil 1.4) were done with Huygens software. We determined the object density for quantification and assessed membrane localization.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Receptor\/protein complexes were detected as well\u2010demarcated 3D objects. Interactions of M2 with Cav\u20101, clathrin, Rab11, and Rab7 were significantly increased in IC\/BPS. M3\/clathrin and M3\/Rab11 complexes were higher in IC\/BPS, while M3\/Cav\u20101 and M3\/Rab7 were not. A significant shift of complexes from the membrane to cytoplasm was observed in conjunction with increased internalization via clathrin vesicles or caveolae in IC\/BPS.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>High numbers of M3\/clathrin and M3\/Rab11 complexes reflect the well\u2010documented clathrin\u2010mediated desensitization of M3 and speak in favor with enhanced receptor protein expression in IC\/BPS. Increased amounts of M2\/Cav\u20101, M2\/clathrin, and M2\/Rab11 complexes represent altered M2 internalization and recycling leading to high abundance in IC\/BPS. In this regard, caveolae\u2010localized M2 could be possibly associated with the activation of nitric oxide (NO) synthase and NO production.<\/jats:p><\/jats:sec>","DOI":"10.1002\/nau.24087","type":"journal-article","created":{"date-parts":[[2019,7,13]],"date-time":"2019-07-13T07:31:07Z","timestamp":1563003067000},"page":"1818-1827","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["IC\/BPS\u2010associated alterations of M2 and M3 muscarinic acetylcholine receptor trafficking in human detrusor"],"prefix":"10.1002","volume":"38","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-2212-2693","authenticated-orcid":false,"given":"Mandy","family":"Berndt\u2010Paetz","sequence":"first","affiliation":[{"name":"Department of Urology, Research Laboratories University of Leipzig  Leipzig 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