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Neurotransmitter release is triggered, in part, by activation of nicotinic acetylcholine receptors (nAChRs). However, despite decades of use as a model system for studying exocytosis, the nAChR subtypes involved have not been pharmacologically identified. Quantitative real\u2010time PCR of rat adrenal medulla revealed an abundance of mRNAs for \u03b13, \u03b17, \u03b22, and \u03b24 subunits. Whole\u2010cell patch\u2010clamp electrophysiology of chromaffin cells and subtype\u2010selective ligands were used to probe for nAChRs derived from the mRNAs found in adrenal medulla. A novel conopeptide antagonist, PeIA\u20105469, was created that is highly selective for \u03b13\u03b22 over other nAChR subtypes heterologously expressed in <jats:italic>Xenopus laevis<\/jats:italic> oocytes. Experiments using PeIA\u20105469 and the \u03b13\u03b24\u2010selective \u03b1\u2010conotoxin TxID revealed that rat adrenal medulla contain two populations of chromaffin cells that express either \u03b13\u03b24 nAChRs alone or \u03b13\u03b24 together with the \u03b13\u03b22\u03b24 subtype. Conclusions were derived from observations that acetylcholine\u2010gated currents in some cells were sensitive to inhibition by PeIA\u20105469 and TxID, while in other cells, currents were sensitive only to TxID. Expression of functional \u03b17 nAChRs was determined using three \u03b17\u2010selective ligands: the agonist PNU282987, the positive allosteric modulator PNU120596, and the antagonist \u03b1\u2010conotoxin [V11L,V16D]ArIB. The results of these studies identify for the first time the expression of \u03b13\u03b22\u03b24 nAChRs as well as functional \u03b17 nAChRs by rat adrenal chromaffin cells.<\/jats:p><\/jats:sec><jats:sec><jats:label\/><jats:p>\n<jats:boxed-text content-type=\"graphic\" position=\"anchor\"><jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc14966-fig-0010-m.png\"><jats:alt-text>image<\/jats:alt-text><\/jats:graphic><\/jats:boxed-text>\n<\/jats:p><\/jats:sec>","DOI":"10.1111\/jnc.14966","type":"journal-article","created":{"date-parts":[[2020,1,22]],"date-time":"2020-01-22T13:02:06Z","timestamp":1579698126000},"page":"158-176","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["Expression of \u03b13\u03b22\u03b24 nicotinic acetylcholine receptors by rat adrenal chromaffin cells determined using novel conopeptide antagonists"],"prefix":"10.1111","volume":"154","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9912-0955","authenticated-orcid":false,"given":"Arik J.","family":"Hone","sequence":"first","affiliation":[{"name":"George E. Whalen Veterans Affairs Medical Center Salt Lake City Utah USA"},{"name":"School of Biological Sciences and University of Utah Salt Lake City Utah USA"},{"name":"Departament of Pharmacology and Therapeutics Universidad Aut\u00f3noma de Madrid Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lola","family":"Rueda\u2010Ruzafa","sequence":"additional","affiliation":[{"name":"Departament of Pharmacology and Therapeutics Universidad Aut\u00f3noma de Madrid Madrid Spain"},{"name":"Biomedical Research Center (CINBIO) University of Vigo Vigo Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas J.","family":"Gordon","sequence":"additional","affiliation":[{"name":"School of Biological Sciences and University of Utah Salt Lake City Utah USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Joanna","family":"Gajewiak","sequence":"additional","affiliation":[{"name":"School of Biological Sciences and University of Utah Salt Lake City Utah USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sean","family":"Christensen","sequence":"additional","affiliation":[{"name":"School of Biological Sciences and University of Utah Salt Lake City Utah USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tino","family":"Dyhring","sequence":"additional","affiliation":[{"name":"Saniona A\/S Ballerup Denmark"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3315-9715","authenticated-orcid":false,"given":"Almudena","family":"Albillos","sequence":"additional","affiliation":[{"name":"Departament of Pharmacology and Therapeutics Universidad Aut\u00f3noma de Madrid Madrid Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J. 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