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Here we have investigated the involvement of H<jats:sub>2<\/jats:sub>S as possible mediator released downstream following muscarinic (M) activation, by using human bladder and urothelial T24 cell line. Carbachol stimulation enhances H<jats:sub>2<\/jats:sub>S production and in turn cGMP in human urothelium or in T24 cells. This effect is reversed by cysthationine-\u03b2-synthase (CBS) inhibition. The blockade of M1 and M3 receptors reverses the increase in H<jats:sub>2<\/jats:sub>S production in human urothelium. In T24 cells, the blockade of M1 receptor significantly reduces carbachol-induced H<jats:sub>2<\/jats:sub>S production. In the functional studies, the urothelium removal from human bladder strips leads to an increase in carbachol-induced contraction that is mimicked by CBS inhibition. Instead, the CSE blockade does not significantly affect carbachol-induced contraction. The increase in H<jats:sub>2<\/jats:sub>S production and in turn of cGMP is driven by CBS-cGMP\/PKG-dependent phosphorylation at Ser<jats:sup>227<\/jats:sup> following carbachol stimulation. The finding of the presence of this crosstalk between the cGMP\/PKG and H<jats:sub>2<\/jats:sub>S pathway downstream to the M1\/M3 receptor in the human urothelium further implies a key role for H<jats:sub>2<\/jats:sub>S in bladder physiopathology. Thus, the modulation of the H<jats:sub>2<\/jats:sub>S pathway can represent a feasible therapeutic target to develop drugs for bladder disorders.<\/jats:p>","DOI":"10.1038\/srep31491","type":"journal-article","created":{"date-parts":[[2016,8,11]],"date-time":"2016-08-11T09:14:25Z","timestamp":1470906865000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":38,"title":["Urothelium muscarinic activation phosphorylates CBSSer227 via cGMP\/PKG pathway causing human bladder relaxation through H2S production"],"prefix":"10.1038","volume":"6","author":[{"given":"Roberta","family":"d\u2019Emmanuele di Villa Bianca","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Mitidieri","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ferdinando","family":"Fusco","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annapina","family":"Russo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Valentina","family":"Pagliara","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Teresa","family":"Tramontano","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erminia","family":"Donnarumma","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Vincenzo","family":"Mirone","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giuseppe","family":"Cirino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Giulia","family":"Russo","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Raffaella","family":"Sorrentino","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2016,8,11]]},"reference":[{"key":"BFsrep31491_CR1","doi-asserted-by":"publisher","first-page":"F103","DOI":"10.1152\/ajprenal.00098.2005","volume":"290","author":"JM Beckel","year":"2006","unstructured":"Beckel, J. 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