{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T19:48:34Z","timestamp":1767988114720,"version":"3.49.0"},"reference-count":31,"publisher":"The Company of Biologists","license":[{"start":{"date-parts":[[2021,5,18]],"date-time":"2021-05-18T00:00:00Z","timestamp":1621296000000},"content-version":"am","delay-in-days":1233,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0"}],"funder":[{"DOI":"10.13039\/501100001807","name":"Funda\u00e9\u00e3o de Amparo \u00e0 Pesquisa do Estado de S\u00e3o Paulo","doi-asserted-by":"publisher","award":["2013\/10780-3"],"award-info":[{"award-number":["2013\/10780-3"]}],"id":[{"id":"10.13039\/501100001807","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003593","name":"Conselho Nacional de Desenvolvimento Cient\u00edfico e Tecnol\u00f3gico","doi-asserted-by":"crossref","award":["147216\/2014-1"],"award-info":[{"award-number":["147216\/2014-1"]}],"id":[{"id":"10.13039\/501100003593","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:p>Xenopus laevis oocytes are a valuable tool for investigating the function of membrane proteins. However, regulations around the world, specifically in Brazil, render the import of Xenopus laevis frogs impractical, and, in some cases, impossible. Here, as an alternative, we evaluate the usefulness of the North American aquatic bullfrog Lithobates catesebeianus, which is commercially available in Brazil, for the heterologous expression of aquaporin (AQP) proteins. We have developed a method that combines a brief collagenase treatment and mechanical defolliculation for isolating individual oocytes from Lithobates ovaries. We find that they have a similar size, shape, and appearance to Xenopus oocytes and can tolerate and survive following injections with cRNA or water. Furthermore, surface biotinylation, western blot analysis, and measurements of osmotic water permeability (Pf) show that Lithobates oocytes can express AQPs to the plasma membrane and significantly increase the Pf of the oocytes. In fact, the Pf values are similar to historical values gathered from Xenopus oocytes. Due to the presence of a mercury sensitive cysteine (Cys or C) in the throat of the water channel, the Pf of oocytes expressing human (h) AQP1, hAQP1FLAG [FLAG, short protein tag (DYKDDDDK) added to the N-terminus of AQP1], hAQP8, and rat (r) AQP9 was inhibited with the mercurial compound p-chloromercuribenzene sulfonate (pCMBS), whereas AQPs lacking this Cys\u2014hAQP1C189S mutant [residue Cys 189 was replaced by a serine (Ser or S)] and hAQP7\u2014were mercury insensitive. Contrary to previous studies with Xenopus oocytes, rAQP3 was also found to be insensitive to mercury, which is consistent with the mercury sensitive Cys (Cys 11) being located intracellularly. Thus, we consider Lithobates oocytes to be a readily accessible system for the functional expression and study of membrane proteins for international researchers who do not currently have access to Xenopus oocytes.<\/jats:p>","DOI":"10.1242\/bio.031880","type":"journal-article","created":{"date-parts":[[2018,3,13]],"date-time":"2018-03-13T00:20:13Z","timestamp":1520900413000},"source":"Crossref","is-referenced-by-count":7,"title":["<i>Lithobates catesbeianus<\/i>(American Bullfrog) oocytes: a novel heterologous expression system for aquaporins"],"prefix":"10.1242","author":[{"given":"J.","family":"Kabutomori","sequence":"first","affiliation":[{"name":"Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, 05508-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"O.","family":"Beloto-Silva","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, 05508-900, Brazil"},{"name":"Paulista University (UNIP), Sao Paulo, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R. R.","family":"Geyer","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, Institute of Chemistry, University of Sao Paulo, Sao Paulo, 05513-970, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"Musa-Aziz","sequence":"additional","affiliation":[{"name":"Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of Sao Paulo, Sao Paulo, 05508-900, Brazil"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","published-online":{"date-parts":[[2018,1,1]]},"reference":[{"key":"2021042423390787400_BIO031880C1","article-title":"Xenopus oocytes: single cell model to study insect olfactory receptors","author":"Batra","year":"2016","journal-title":"NICE"},{"key":"2021042423390787400_BIO031880C2","doi-asserted-by":"publisher","first-page":"127","DOI":"10.2994\/057.006.0203","article-title":"Widespread occurrence of the American Bullfrog, Lithobates catesbeianus (Shaw, 1802) (Anura: Ranidae), in Brazil","volume":"6","author":"Both","year":"2011","journal-title":"South Am. 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