Apoptosis of Alcohol‐Exposed Human Placental Cytotrophoblast Cells is Downstream of Intracellular Calcium Signaling. (29th May 2014)

• Record Type: Journal Article
• Authors: Bolnick, Jay M.; Karana, Rita; Chiang, Po J.; Kilburn, Brian A.; Romero, Roberto; Diamond, Michael P.; Smith, Susan M.; Armant, D. Randall
• Is Part Of: Alcoholism. Volume 38:Number 6(2014:Jun.)
• Page Start: 1646
• Page End: 1653
• Publication Date: 2014-05-29
• Subjects: Alcoholism -- Periodicals; Alcoholism -- Periodicals; Alcoolisme; Electronic journals; Périodique électronique (Descripteur de forme); Ressource Internet (Descripteur de forme); 616.861005
• DOI: 10.1111/acer.12417
• Languages: English
• ISSNs: 0145-6008

Abstract

<abstract abstract-type="main" id="acer12417-abs-0001"> <title> <x xml:space="preserve">Abstract</x> </title> <sec id="acer12417-sec-0001" sec-type="section"> <title>Background</title> <p>Apoptosis is induced by ethanol (EtOH) in human placental trophoblast cells, possibly disrupting placentation and contributing to intrauterine growth restriction in fetal alcohol spectrum disorder (FASD). EtOH induces programmed cell death in several embryonic tissues by raising intracellular Ca<sup>2+</sup>. Therefore, the role of Ca<sup>2+</sup> signaling in EtOH‐induced apoptosis was examined using human first trimester cytotrophoblast cell lines, examining the hypothesis that apoptosis is dependent on intracellular Ca<sup>2+</sup> signaling.</p> </sec> <sec id="acer12417-sec-0002" sec-type="section"> <title>Methods</title> <p>Using HTR‐8/SVneo and SW.71 cytotrophoblast cell lines, real‐time intracellular Ca<sup>2+</sup> concentration was monitored by fluo‐4 epifluorescence microscopy and apoptosis was assessed by flow cytometry of cells fluorescently labeled for DNA fragmentation (TUNEL) and annexin V binding.</p> </sec> <sec id="acer12417-sec-0003" sec-type="section"> <title>Results</title> <p>Intracellular Ca<sup>2+</sup> concentrations increased synchronously in all cells within 10 seconds of exposure to 50 mM EtOH, but not at lower EtOH concentrations (10 to 25 mM) incapable of inducing apoptosis. Trophoblast cells treated with inhibitors of Ca<sup>2+</sup> signaling (BAPTA‐AM, U73122, xestospongin D, BAPTA, SKF‐96365) produced no intracellular Ca<sup>2+</sup> transients after exposure to 50 mM EtOH and were protected from cell death induced by EtOH.</p> </sec> <sec id="acer12417-sec-0004" sec-type="section"> <title>Conclusions</title> <p>EtOH‐induced apoptosis in human cytotrophoblast cells, identified by DNA fragmentation and externalized phosphatidylserine, was dependent upon Ca<sup>2+</sup> signaling. Both intracellular Ca<sup>2+</sup> mobilization and extracellular Ca<sup>2+</sup> influx were required, as well as phosphatidylinositol signaling. Inhibition by SKF‐96365 suggests that the capacitative Ca<sup>2+</sup> entry mechanism that utilizes TRPC channels was activated by EtOH. Apoptosis occurs downstream of Ca<sup>2+</sup> signaling in trophoblasts and may contribute to placental insufficiency and poor fetal growth associated with FASD.</p> </sec> </abstract>