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Drug Alcohol Res."],"abstract":"<jats:p>Despite the significant number of people who may be taking pregnenolone supplements while drinking alcohol (ethanol), the widely documented cerebrovascular actions of pregnenolone and ethanol, and the critical dependence of cerebrovascular function on cerebral artery diameter, there are no studies addressing the effect of pregnenolone + ethanol in combination on cerebral artery diameter. We investigated this by evaluating the effect of this combination on middle cerebral artery diameter in male and female C57BL\/6J mice, both <jats:italic>in vivo<\/jats:italic> and <jats:italic>in vitro<\/jats:italic>. The use of de-endothelialized, <jats:italic>in vitro<\/jats:italic> pressurized middle cerebral artery segments allowed us to conduct a concentration-response study of constriction induced by pregnenolone \u00b1 ethanol, in which drug action could be evaluated independently of circulating and endothelial factors. In both male and female animals, pregnenolone at lower concentrations (\u22641\u00a0\u00b5M) was found to synergize with 50\u00a0mM ethanol to cause vasoconstriction. In both sexes, this synergism was lost as one or both vasoconstrictors approached their maximally effective concentrations (75\u00a0mM and 10\u00a0\u00b5M for ethanol and pregnenolone, respectively), whether this was evaluated <jats:italic>in vitro<\/jats:italic> or <jats:italic>in vivo<\/jats:italic> using a cranial window. Vasoconstriction by pregnenolone + ethanol was abolished by 1\u00a0\u00b5M paxilline, indicating BK channel involvement. Moreover, cell-free recordings of BK channel activity in cerebral artery myocyte membranes showed that 10\u00a0\u00b5M pregnenolone and pregnenolone +50\u00a0mM ethanol reduced channel activity to an identical extent, suggesting that these drugs inhibit cerebrovascular BK channels <jats:italic>via<\/jats:italic> a common mechanism or mechanisms. Indeed, pregnenolone was found to disrupt allosteric coupling to Ca<jats:sup>2+<\/jats:sup>-driven gating, as previously reported for ethanol.<\/jats:p>","DOI":"10.3389\/adar.2023.11735","type":"journal-article","created":{"date-parts":[[2023,8,14]],"date-time":"2023-08-14T04:11:20Z","timestamp":1691986280000},"update-policy":"https:\/\/doi.org\/10.3389\/crossmark-policy","source":"Crossref","is-referenced-by-count":0,"title":["Alcohol and pregnenolone interaction on cerebral arteries through targeting of vascular smooth muscle Ca2+- and voltage-gated K+ channels of big conductance"],"prefix":"10.3389","volume":"3","author":[{"given":"Kelsey C.","family":"North","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew A.","family":"Shaw","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"Jr.","given":"Luiz","family":"Moreira","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anna N.","family":"Bukiya","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alex M.","family":"Dopico","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1965","published-online":{"date-parts":[[2023,8,14]]},"reference":[{"key":"B1","doi-asserted-by":"publisher","first-page":"1017","DOI":"10.1037\/e627412011-001","article-title":"Alcohol misuse","volume":"2011","author":"Coulton","year":"2011","journal-title":"BMJ Clin Evid"},{"key":"B2","unstructured":"Binge drinking is a serious but preventable public health problem2021"},{"volume-title":"Center for behavioral health statistics and quality","year":"2021","key":"B3"},{"key":"B4","doi-asserted-by":"publisher","first-page":"717","DOI":"10.1016\/j.amepre.2017.02.014","article-title":"Drinking beyond the binge threshold: predictors, consequences, and changes in the U.S","volume":"52","author":"Hingson","year":"2017","journal-title":"Am J Prev Med"},{"volume-title":"Center for behavioral statistics and quality","year":"2019","key":"B5"},{"key":"B6","doi-asserted-by":"publisher","first-page":"315","DOI":"10.1111\/j.1360-0443.1995.tb03779.x","article-title":"Alcohol amnesia","volume":"90","author":"Goodwin","year":"1995","journal-title":"Addiction"},{"key":"B7","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1080\/07448480209595713","article-title":"Trends in college binge drinking during a period of increased prevention efforts. 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