Evaluation of the role of nitric oxide in acid sensing ion channel mediated cell death.
Acid sensing ion channels (ASICs) are widely expressed in central and peripheral nervous system. They are involved in a variety of physiological and pathophysiological processes: synaptic transmission, learning and memory, pain perception, ischemia, etc. During ischemia, metabolic acidosis causes the drop of extracellular pH (pHe) which in turn activates ASICs. Activation of calcium permeable ASIC1a has been implicated in neuronal death. ASICs are modulated by several redox reagents, divalent cations and nitric oxide (NO). Although NO potentiates ASIC mediated currents, the physiological significance of such modulation has not been studied in detail. We have evaluated the role of endogenous NO in cell death at different pH, mediated by the activation of ASICs. At pH 6.1, death rates of ASIC1 expressing Neuro2A (N2A) cells are significantly higher in comparison to the cells that do not express ASICs. Amiloride, a blocker of ASICs protects the cell from acid-injury. Sodium nitroprusside, a potent NO donor not only increases the ASIC mediated currents but also increases cell death at low pH. L-Arg, the precursor of NO also potentiates ASICs in a pH dependent manner. L-Arg-induced NO production and potentiation of ASICs were observed at pHs 7.4, 7.2, 7.0 and 6.8. Lowering the pH below 6.8 did not result in significant production of NO or potentiation of ASICs upon L-Arg stimulation. Our results suggest that potentiation of ASICs by NO and subsequent cell death in vivo depends on the severity of acidosis. During mild and moderate acidosis, NO promotes cell death by potentiating ASICs, whereas this potentiation subsides in severe acidosis due to inhibition of NO synthase.
Jetti
Suresh Kumar
SK
Department of Biotechnology, IIT Madras, Chennai, India.
Swain
Sandip Madhusudan
SM
Majumder
Syamantak
S
Chatterjee
Suvro
S
Poornima
V
V
Bera
Amal Kanti
AK
D062565:Acid Sensing Ion Channels; D000143:Acids / Q000494:pharmacology; D000818:Animals; D016466:CHO Cells; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D006367:HeLa Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D009569:Nitric Oxide / Q000032:analysis / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009599:Nitroprusside / Q000494:pharmacology; D051381:Rats; D017208:Rats, Wistar; D015222:Sodium Channels / Q000378:metabolism*
C568266:ASIC1 protein, human; D062565:Acid Sensing Ion Channels; D000143:Acids; C568268:Asic1 protein, rat; D009419:Nerve Tissue Proteins; D020030:Nitric Oxide Donors; D015222:Sodium Channels; D009599:Nitroprusside; D009569:Nitric Oxide