Acidosis and 5-(N-ethyl-N-isopropyl)amiloride (EIPA) Attenuate Zinc/Kainate Toxicity in Cultured Cerebellar Granule Neurons

Biochemistry (Mosc). 2015 Aug;80(8):1065-72. doi: 10.1134/S000629791508012X.

Abstract

Cultured cerebellar granule neurons (CGNs) are resistant to the toxic effect of ZnCl2 (0.005 mM, 3 h) and slightly sensitive to the effect of kainate (0.1 mM, 3 h). Simultaneous treatment of CGNs with kainate and ZnCl2 caused intensive neuronal death, which was attenuated by external acidosis (pH 6.5) or 5-(N-ethyl-N-isopropyl)amiloride (EIPA, Na+/H+ exchange blocker, 0.03 mM). Intracellular zinc and calcium ion concentrations ([Zn2+]i and [Ca2+]i) were increased under the toxic action of kainate + ZnCl2, this effect being significantly decreased on external acidosis and increased in case of EIPA addition. Neuronal Zn2+ imaging demonstrated that EIPA increases the cytosolic concentration of free Zn2+ on incubation in Zn2+-containing solution. These data imply that acidosis reduces ZnCl2/kainate toxic effects by decreasing Zn2+ entry into neurons, and EIPA prevents zinc stores from being overloaded with zinc.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acidosis / metabolism*
  • Amiloride / analogs & derivatives*
  • Amiloride / pharmacology
  • Animals
  • Calcium / pharmacology
  • Cations, Divalent
  • Cell Death / drug effects
  • Cells, Cultured
  • Cerebellum / cytology
  • Cerebellum / drug effects
  • Cytoplasmic Granules / pathology
  • Drug Interactions
  • Kainic Acid / metabolism
  • Kainic Acid / toxicity
  • Neurons / drug effects*
  • Neurons / metabolism*
  • Rats
  • Rats, Wistar
  • Receptors, AMPA / metabolism
  • Receptors, Kainic Acid / metabolism
  • Sodium-Hydrogen Exchangers / metabolism
  • Zinc / metabolism
  • Zinc / toxicity

Substances

  • Cations, Divalent
  • Receptors, AMPA
  • Receptors, Kainic Acid
  • Sodium-Hydrogen Exchangers
  • Amiloride
  • Zinc
  • Kainic Acid
  • Calcium
  • ethylisopropylamiloride