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Journal of Neuroscience, Vol 16, 2002-2011, Copyright © 1996 by Society for Neuroscience
Reduced nicotinamide adenine dinucleotide-selective stimulation of inositol 1,4,5-trisphosphate receptors mediates hypoxic mobilization of calcium
AI Kaplin, SH Snyder and DJ Linden
Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
To evaluate the relationship of inositol 1,4,5-trisphosphate (IP3)
receptor-mediated signal transduction and cellular energy dynamics, we have
characterized effects of nucleotides on IP3 receptor (IP3R)- mediated
calcium (Ca2+) flux in purified IP3 receptors reconstituted in lipid
vesicles (IP3RV) and examined hypoxia-induced augmentation of intracellular
Ca2+ in intact cells. Reduced nicotinamide adenine dinucleotide (NADH)
increases IP3-mediated Ca2+ flux in IP3RV. This effect is highly specific
for NADH. Hypoxia elicited by brief exposure of nerve growth
factor-differentiated PC12 cells or cerebellar Purkinje cells to cyanide
elicits rapid increased in internal [Ca2+], which derives from
IP3-sensitive stores. Blockade of this effect by 2- deoxyglucose and
inhibition of glyceraldehyde-3-phosphate dehydrogenase implicates enhanced
glycolytic production of NADH in the Ca2+ stimulation. Internal [Ca2+] is
markedly and specifically increased by direct intracellular injection of
NADH, and this effect is blocked by heparin, further implicating IP3R
stores. These findings indicate that direct regulation of IP3R by NADH is
responsible for elevated cytoplasmic [Ca2+] occurring in the earliest phase
of hypoxia. This link of IP3R activity with cellular energy dynamics may be
relevant to both hypoxic damage and metabolic regulation of IP3 signaling
processes.
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