{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,11]],"date-time":"2024-07-11T02:18:05Z","timestamp":1720664285553},"reference-count":31,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2013,2,19]],"date-time":"2013-02-19T00:00:00Z","timestamp":1361232000000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Alcoholism Clin &amp;amp; Exp Res"],"published-print":{"date-parts":[[2013,6]]},"abstract":"<jats:sec><jats:title>Background<\/jats:title><jats:p>Voltage\u2010gated calcium channels (<jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content>s) in rat neurohypophysial terminals exhibit molecular tolerance to alcohol, including desensitization to the drug and increased current density, after 3\u00a0weeks of alcohol drinking. Moreover, after this time, terminals from drinking rats exhibit diminished alcohol inhibition of vasopressin (<jats:styled-content style=\"fixed-case\">AVP<\/jats:styled-content>) release.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>We took advantage of organotypic cultures (explants) of the hypothalamo\u2010neurohypophysial system (<jats:styled-content style=\"fixed-case\">HNS<\/jats:styled-content>) to extend our analysis of molecular tolerance to 2 classes of the <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content>. The isolated <jats:styled-content style=\"fixed-case\">HNS<\/jats:styled-content> explant allows much finer temporal resolution of molecular tolerance than do voluntary drinking paradigms. After exposure of the <jats:styled-content style=\"fixed-case\">HNS<\/jats:styled-content> explant to alcohol, terminals are isolated by mechanical treatment and plated in a dish. Patch clamp recording techniques are used to obtain <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content> currents, and immunohistochemistry is used to determine <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content> distribution. A release assay is used to provide functional readout of <jats:styled-content style=\"fixed-case\">AVP<\/jats:styled-content> release.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>We show that even a brief, 1\u2010hour exposure to a clinically relevant concentration of alcohol is sufficient to evoke similar changes to those observed after several weeks of exposure. Acute ethanol (<jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>t<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content>) exposure inhibits high <jats:styled-content style=\"fixed-case\"><jats:roman>K<\/jats:roman><\/jats:styled-content><jats:sup>+<\/jats:sup>\u2010induced <jats:styled-content style=\"fixed-case\">AVP<\/jats:styled-content> release from na\u00efve terminals. However, terminals pre\u2010exposed to 20\u00a0mM <jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>t<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content> for 1\u00a0hour become tolerant to <jats:styled-content style=\"fixed-case\">E<\/jats:styled-content>t<jats:styled-content style=\"fixed-case\">OH<\/jats:styled-content>, and subsequent exposure has significantly less effect on high <jats:styled-content style=\"fixed-case\"><jats:roman>K<\/jats:roman><\/jats:styled-content><jats:sup>+<\/jats:sup>\u2010induced <jats:styled-content style=\"fixed-case\">AVP<\/jats:styled-content> release. Electrophysiological recordings indicate that among different types of <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content>s present in the neuronal terminal, the <jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>\u2010type is the most affected by alcohol. The current density of <jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>\u2010type current is significantly increased (approximately 50%), while its responsiveness to alcohol is significantly diminished (approximately 50%), after brief alcohol exposure. Fluorescent imaging results were consistent with the electrophysiology and suggest that the increased current density of <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content>s after brief exposure is attributable to combined synthesis of 1.2 and 1.3 subtypes of the <jats:styled-content style=\"fixed-case\">L<\/jats:styled-content>\u2010type <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content> and redistribution of channel protein into terminal plasma membrane.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>These data indicate that a brief alcohol exposure affects subsequent alcohol sensitivity of <jats:styled-content style=\"fixed-case\">VGCC<\/jats:styled-content>s and neuropeptide release from presynaptic terminals.<\/jats:p><\/jats:sec>","DOI":"10.1111\/acer.12057","type":"journal-article","created":{"date-parts":[[2013,2,19]],"date-time":"2013-02-19T22:46:59Z","timestamp":1361314019000},"page":"933-940","source":"Crossref","is-referenced-by-count":7,"title":["Molecular Tolerance of Voltage\u2010Gated Calcium Channels is Evident After Short Exposures to Alcohol in Vasopressin\u2010Releasing Nerve Terminals"],"prefix":"10.1111","volume":"37","author":[{"given":"Andrzej Z.","family":"Pietrzykowski","sequence":"first","affiliation":[{"name":"Department of Animal Sciences  Rutgers University New Brunswick New Jersey"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sonia","family":"Ortiz\u2010Miranda","sequence":"additional","affiliation":[{"name":"Department of MaPS &amp; Program in Neuroscience  University of Massachusetts Medical School Worcester Massachusetts"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Thomas 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