Intracellular calcium plays a critical role in the alcohol-mediated death of cerebellar granule neurons

J Neurochem. 2013 Feb;124(3):323-35. doi: 10.1111/jnc.12076. Epub 2012 Dec 6.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Calcium / physiology*
  • Calcium Signaling / drug effects
  • Calcium Signaling / physiology
  • Cell Death / drug effects
  • Cell Death / physiology
  • Cells, Cultured
  • Central Nervous System Depressants / toxicity
  • Cerebellum / drug effects
  • Cerebellum / metabolism
  • Cerebellum / pathology*
  • Culture Media / pharmacology
  • Cytoplasmic Granules / drug effects
  • Cytoplasmic Granules / physiology
  • Ethanol / toxicity*
  • Female
  • Fetal Alcohol Spectrum Disorders / metabolism*
  • Fetal Alcohol Spectrum Disorders / pathology*
  • Intracellular Space / drug effects
  • Intracellular Space / physiology
  • Male
  • Mice
  • Mice, 129 Strain
  • Mice, Inbred C57BL
  • Neurons / drug effects
  • Neurons / metabolism
  • Neurons / pathology*
  • Pregnancy

Substances

  • Central Nervous System Depressants
  • Culture Media
  • Ethanol
  • Calcium