{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T01:22:06Z","timestamp":1791249726282,"version":"4.1.0"},"reference-count":48,"publisher":"Cambridge University Press (CUP)","issue":"1","license":[{"start":{"date-parts":[[2013,7,22]],"date-time":"2013-07-22T00:00:00Z","timestamp":1374451200000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Zygote"],"published-print":{"date-parts":[[2015,2]]},"abstract":"<jats:title>Summary<\/jats:title><jats:p>Studies on bovine oocytes have revealed that the activation of adenosine monophosphate activated protein kinase (AMPK) by millimolar concentrations of metformin controls nuclear maturation. Tuberous sclerosis complex 2 (TSC2) has been identified as a downstream target of AMPK. The objective of this study was to investigate the effects of addition of low concentrations of metformin (1 nM to 10 \u03bcM) on the percentage of cultured cumulus\u2013oocyte complexes (COC) giving rise to cleavage-stage embryos and AMPK-mediated TSC2 activation. Metformin was supplemented either throughout <jats:italic>in vitro<\/jats:italic> embryo production (IVP) or only during <jats:italic>in vitro<\/jats:italic> fertilization (IVF). COC were matured <jats:italic>in vitro<\/jats:italic>, inseminated, and presumptive zygotes cultured for a further 72 h post insemination before the percentage of COC that gave rise to zygotes and early embryo development was assessed. The presence of TSC2 in bovine embryos and its possible AMPK-induced activation were assessed by immunocytochemistry. Metformin had a dose-dependent effect on the numbers of cultured COC that gave rise to embryos. Drug treatment either throughout IVP or only during IVF decreased the percentage of \u22658-cell embryos (1 \u03bcM, <jats:italic>P<\/jats:italic> &lt; 0.05; 10 \u03bcM, <jats:italic>P<\/jats:italic> &lt; 0.01; and 0.1 \u03bcM, 10 \u03bcM, <jats:italic>P<\/jats:italic> &lt; 0.01, respectively) and increased the percentage of 2-cell embryos (10 \u03bcM, <jats:italic>P<\/jats:italic> &lt; 0.01 and <jats:italic>P<\/jats:italic> &lt; 0.05 respectively). The percentage of cultured COC that gave rise to zygotes was not affected by metformin. TSC2 is expressed in early embryos. Metformin (10 \u03bcM) either throughout IVP or during IVF only, increased AMPK-induced Phospho<jats:sup>S1387<\/jats:sup>-TSC2 immunoreactivity (<jats:italic>P<\/jats:italic> &lt; 0.01) and this increase corresponded to the total TSC2 protein levels expressed in cells. Our results suggest that there is a dose-dependent negative effect of metformin on the ability of oocytes to cleave following insemination, possibly mediated through an AMPK-induced activation of TSC2.<\/jats:p>","DOI":"10.1017\/s0967199413000300","type":"journal-article","created":{"date-parts":[[2013,7,22]],"date-time":"2013-07-22T10:24:41Z","timestamp":1374488681000},"page":"58-67","source":"Crossref","is-referenced-by-count":12,"title":["Effects of metformin on fertilisation of bovine oocytes and early embryo development: possible involvement of AMPK3-mediated TSC2 activation"],"prefix":"10.1017","volume":"23","author":[{"given":"Olympia","family":"Pikiou","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Vasilaki","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"George","family":"Leondaritis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nikos","family":"Vamvakopoulos","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ioannis E.","family":"Messinis","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"56","published-online":{"date-parts":[[2013,7,22]]},"reference":[{"key":"S0967199413000300_ref36","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2004.06.007"},{"key":"S0967199413000300_ref32","doi-asserted-by":"publisher","DOI":"10.1038\/nrc1974"},{"key":"S0967199413000300_ref19","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)80657-1"},{"key":"S0967199413000300_ref41","doi-asserted-by":"publisher","DOI":"10.1210\/en.2005-0301"},{"key":"S0967199413000300_ref38","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.271.2.611"},{"key":"S0967199413000300_ref39","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod64.3.904"},{"key":"S0967199413000300_ref17","doi-asserted-by":"publisher","DOI":"10.2165\/00003495-200363180-00001"},{"key":"S0967199413000300_ref34","doi-asserted-by":"publisher","DOI":"10.1126\/science.1157535"},{"key":"S0967199413000300_ref16","doi-asserted-by":"publisher","DOI":"10.1017\/S0967199409005255"},{"key":"S0967199413000300_ref14","doi-asserted-by":"publisher","DOI":"10.1111\/j.1440-1681.2008.05131.x"},{"key":"S0967199413000300_ref13","doi-asserted-by":"publisher","DOI":"10.1111\/j.1432-1033.1997.00259.x"},{"key":"S0967199413000300_ref40","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(01)00332-4"},{"key":"S0967199413000300_ref47","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.01.016"},{"key":"S0967199413000300_ref45","doi-asserted-by":"publisher","DOI":"10.1161\/01.RES.0000256090.42690.05"},{"key":"S0967199413000300_ref21","doi-asserted-by":"publisher","DOI":"10.1530\/REP-09-0517"},{"key":"S0967199413000300_ref43","doi-asserted-by":"publisher","DOI":"10.1677\/joe.1.06828"},{"key":"S0967199413000300_ref12","first-page":"214","article-title":"AMPK phosphorylation of Raptor mediates a metabolic checkpoint","volume":"30","author":"Gwinn","year":"2008","journal-title":"Cell"},{"key":"S0967199413000300_ref35","doi-asserted-by":"publisher","DOI":"10.1038\/nrc2676"},{"key":"S0967199413000300_ref31","doi-asserted-by":"crossref","unstructured":"Potter C.J. & Xu T. (2001). Mechanisms of size control. Curr Opin Genet Dev 11, 279\u2013477.","DOI":"10.1016\/S0959-437X(00)00191-X"},{"key":"S0967199413000300_ref9","doi-asserted-by":"publisher","DOI":"10.1101\/gad.901101"},{"key":"S0967199413000300_ref25","doi-asserted-by":"publisher","DOI":"10.1016\/j.tibs.2003.09.003"},{"key":"S0967199413000300_ref42","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.106.050831"},{"key":"S0967199413000300_ref46","doi-asserted-by":"publisher","DOI":"10.1167\/iovs.03-0835"},{"key":"S0967199413000300_ref48","doi-asserted-by":"publisher","DOI":"10.1172\/JCI13505"},{"key":"S0967199413000300_ref10","doi-asserted-by":"publisher","DOI":"10.1111\/j.1439-0531.2011.01770.x"},{"key":"S0967199413000300_ref8","doi-asserted-by":"publisher","DOI":"10.2337\/db07-0074"},{"key":"S0967199413000300_ref11","doi-asserted-by":"publisher","DOI":"10.1016\/j.ccr.2007.05.008"},{"key":"S0967199413000300_ref7","doi-asserted-by":"publisher","DOI":"10.1006\/dbio.2002.0613"},{"key":"S0967199413000300_ref30","first-page":"373","article-title":"Metformin therapy in the management of polycystic ovary syndrome: endocrine, metabolic and reproductive effects","volume":"15","author":"Nardo","year":"2001","journal-title":"Gynecol. Endocrinol."},{"key":"S0967199413000300_ref33","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2007.03.003"},{"key":"S0967199413000300_ref28","doi-asserted-by":"publisher","DOI":"10.1200\/JCO.2008.20.0766"},{"key":"S0967199413000300_ref4","doi-asserted-by":"publisher","DOI":"10.1097\/00060793-199702000-00006"},{"key":"S0967199413000300_ref29","doi-asserted-by":"publisher","DOI":"10.2337\/diabetes.51.7.2074"},{"key":"S0967199413000300_ref1","doi-asserted-by":"publisher","DOI":"10.1530\/rep.1.00871"},{"key":"S0967199413000300_ref24","doi-asserted-by":"publisher","DOI":"10.2217\/fon.09.174"},{"key":"S0967199413000300_ref26","doi-asserted-by":"publisher","DOI":"10.2337\/diab.40.7.850"},{"key":"S0967199413000300_ref18","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(03)00929-2"},{"key":"S0967199413000300_ref3","doi-asserted-by":"publisher","DOI":"10.1007\/s00018-009-0163-7"},{"key":"S0967199413000300_ref27","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.106.057828"},{"key":"S0967199413000300_ref2","doi-asserted-by":"publisher","DOI":"10.1093\/molehr\/gap092"},{"key":"S0967199413000300_ref23","doi-asserted-by":"publisher","DOI":"10.1136\/bmj.327.7421.951"},{"key":"S0967199413000300_ref44","doi-asserted-by":"publisher","DOI":"10.1095\/biolreprod.107.060848"},{"key":"S0967199413000300_ref15","doi-asserted-by":"publisher","DOI":"10.1016\/0093-691X(94)00043-T"},{"key":"S0967199413000300_ref5","doi-asserted-by":"publisher","DOI":"10.1002\/mrd.20574"},{"key":"S0967199413000300_ref20","first-page":"31","article-title":"The pharmacokinetics of metformin: a comparison of the properties of a rapid-release and a sustained release preparation. Int.","volume":"21","author":"Karttunen","year":"1983","journal-title":"J. Clin. Pharmacol."},{"key":"S0967199413000300_ref37","doi-asserted-by":"publisher","DOI":"10.1126\/science.1120781"},{"key":"S0967199413000300_ref6","doi-asserted-by":"publisher","DOI":"10.1101\/gad.1199104"},{"key":"S0967199413000300_ref22","doi-asserted-by":"publisher","DOI":"10.1016\/S1262-3636(03)72792-X"}],"container-title":["Zygote"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.cambridge.org\/core\/services\/aop-cambridge-core\/content\/view\/S0967199413000300","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,4,21]],"date-time":"2019-04-21T04:09:06Z","timestamp":1555819746000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.cambridge.org\/core\/product\/identifier\/S0967199413000300\/type\/journal_article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2013,7,22]]},"references-count":48,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2015,2]]}},"alternative-id":["S0967199413000300"],"URL":"https:\/\/doi.org\/10.1017\/s0967199413000300","relation":{},"ISSN":["0967-1994","1469-8730"],"issn-type":[{"value":"0967-1994","type":"print"},{"value":"1469-8730","type":"electronic"}],"subject":[],"published":{"date-parts":[[2013,7,22]]}}}