ATP and nitric oxide modulate intracellular calcium in isolated pillar cells of the guinea pig cochlea.
Supporting cells in the mammalian cochlea have recently received attention as potential targets of neurotransmitters, neuromodulators, and neurohumoral agents. Calcium homeostasis in Deiters' and Hensen's cells, for example, is regulated by ATP and nitric oxide. We studied the intracellular calcium concentration [Ca2+]i in isolated pillar cells of the guinea pig cochlea in response to extracellular ATP and nitric oxide using the fluorescent indicator fluo-3. [Ca2+]i increased rapidly and significantly throughout the pillar cell in response to a bolus of ATP or 2-methylthio ATP while alpha,beta-methylene ATP was ineffective. The response to ATP was inhibited by suramin and Cibacron Blue but not by pyridoxal phosphate 6-azophenyl-2',4'-disulfonic acid. This pharmacological profile is consistent with a [Ca2+]i increase largely mediated by P2Y receptors. In Ca2+-free medium supplemented with EGTA, the response to extracellularATP was reduced by 33%, suggesting a contribution of calcium influx to the overall effect. The ATP-induced increase of [Ca2+] was attenuated by NO donors (sodium nitroprusside or diethylamine NONOate), and this attenuation was reversed by KT5823, an antagonist to protein kinase G. The results indicate the involvement of purinergic mechanisms and the nitric oxide/cyclic GMP/protein kinase G pathway in the regulation of [Ca2+]i in cochlear pillar cells.
Chung
J W
JW
Kresge Hearing Research Institute, University of Michigan, Ann Arbor 48109-0506, USA.
Schacht
J
J
D000255:Adenosine Triphosphate / Q000494:pharmacology / Q000502:physiology*; D000818:Animals; D002118:Calcium / Q000032:analysis / Q000378:metabolism*; D003051:Cochlea / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D006152:Cyclic GMP / Q000502:physiology; D006168:Guinea Pigs; D007425:Intracellular Membranes / Q000378:metabolism*; D007552:Isotonic Solutions / Q000737:chemistry / Q000494:pharmacology; D008297:Male; D009569:Nitric Oxide / Q000502:physiology*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D009994:Osmolar Concentration; D058913:Purinergic Agonists; D058914:Purinergic Antagonists
C072670:Hanks Balanced Salt Solution; D007552:Isotonic Solutions; D020030:Nitric Oxide Donors; D058913:Purinergic Agonists; D058914:Purinergic Antagonists; D009569:Nitric Oxide; D000255:Adenosine Triphosphate; D006152:Cyclic GMP; D002118:Calcium