title: Intracellular calcium plays a critical role in the alcohol-mediated death of cerebellar granule neurons.
abstract: Alcohol is a potent neuroteratogen that can trigger neuronal death in the developing brain. However, the mechanism underlying this alcohol-induced neuronal death is not fully understood. Utilizing primary cultures of cerebellar granule neurons (CGN), we tested the hypothesis that the alcohol-induced increase in intracellular calcium [Ca(2+)](i) causes the death of CGN. Alcohol induced a dose-dependent (200-800 mg/dL) neuronal death within 24 h. Ratiometric Ca(2+) imaging with Fura-2 revealed that alcohol causes a rapid (1-2 min), dose-dependent increase in [Ca(2+)](i), which persisted for the duration of the experiment (5 or 7 min). The alcohol-induced increase in [Ca(2+)](i) was observed in Ca(2+) -free media, suggesting intracellular Ca(2+) release. Pre-treatment of CGN cultures with an inhibitor (2-APB) of the inositol-triphosphate receptor (IP(3) R), which regulates Ca(2+) release from the endoplasmic reticulum (ER), blocked both the alcohol-induced rise in [Ca(2+)](i) and the neuronal death caused by alcohol. Similarly, pre-treatment with BAPTA/AM, a Ca(2+) -chelator, also inhibited the alcohol-induced surge in [Ca(2+) ](i) and prevented neuronal death. In conclusion, alcohol disrupts [Ca(2+)](i) homeostasis in CGN by releasing Ca(2+) from intracellular stores, resulting in a sustained increase in [Ca(2+)](i). This sustained increase in [Ca(2+)](i) may be a key determinant in the mechanism underlying alcohol-induced neuronal death.
authors.affiliation: Department of Anatomy and Cell Biology, Carver College of Medicine, The University of Iowa, Iowa City, IA, USA.
authors.forename: Dimitrios E
authors.initials: DE
authors.lastname: Kouzoukas
authors.forename: Guiying
authors.initials: G
authors.lastname: Li
authors.forename: Maysaam
authors.initials: M
authors.lastname: Takapoo
authors.forename: Thomas
authors.initials: T
authors.lastname: Moninger
authors.forename: Ramesh C
authors.initials: RC
authors.lastname: Bhalla
authors.forename: Nicholas J
authors.initials: NJ
authors.lastname: Pantazis
mesh_terms: D000818:Animals; D002118:Calcium / Q000502:physiology*; D020013:Calcium Signaling / Q000187:drug effects / Q000502:physiology; D016923:Cell Death / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D002492:Central Nervous System Depressants / Q000633:toxicity; D002531:Cerebellum / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D003470:Culture Media / Q000494:pharmacology; D003594:Cytoplasmic Granules / Q000187:drug effects / Q000502:physiology; D000431:Ethanol / Q000633:toxicity*; D005260:Female; D063647:Fetal Alcohol Spectrum Disorders / Q000378:metabolism* / Q000473:pathology*; D042541:Intracellular Space / Q000187:drug effects / Q000502:physiology; D008297:Male; D051379:Mice; D057507:Mice, 129 Strain; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D011247:Pregnancy
chemical_list: D002492:Central Nervous System Depressants; D003470:Culture Media; D000431:Ethanol; D002118:Calcium