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The Journal of Neuroscience, March 1, 2000, 20(5):1767-1779
A Fundamental Role for the Nitric Oxide-G-Kinase Signaling Pathway in Mediating Intercellular Ca2+ Waves in Glia
Nicholas J. Willmott, Kay Wong, and Anthony J. Strong Department of Clinical Neuroscience, Institute of Psychiatry, King's College London, London SE5 8AF
In this study, we highlight a role for the nitric oxide-cGMP-dependent protein kinase (NO-G-kinase) signaling pathway in glial intercellular Ca2+ wave initiation and propagation. Addition of the NO donor molsidomine (100-500 µM) or puffing aqueous NO onto primary glial cell cultures evoked an increase in [Ca2+]i in individual cells and also local intercellular Ca2+ waves, which persisted after removal of extracellular Ca2+. High concentrations of ryanodine (100-200 µM) and antagonists of the NO-G-kinase signaling pathway essentially abrogated the NO-induced increase in [Ca2+]i, indicating that NO mobilizes Ca2+ from a ryanodine receptor-linked store, via the NO-G-kinase signaling pathway. Addition of 10 µM nicardipine to cells resulted in a slowing of the molsidomine-induced rise in [Ca2+]i, and inhibition of Mn2+ quench of cytosolic fura-2 fluorescence mediated by a bolus application of 2 µM aqueous NO to cells, indicating that NO also induces Ca2+ influx in glia. Mechanical stress of individual glial cells resulted in an increase in intracellular NO in target and neighboring cells and intercellular Ca2+ waves, which were NO, cGMP, and G-kinase dependent, because incubating cells with nitric oxide synthase, guanylate cyclase, and G-kinase inhibitors, or NO scavengers, reduced
[Ca2+]i and the rate of Ca2+ wave propagation in these cultures. Results from this study suggest that NO-G-kinase signaling is coupled to Ca2+ mobilization and influx in glial cells and that this pathway plays a fundamental role in the generation and propagation of intercellular Ca2+ waves in glia.
Key words: nitric oxide; glia; calcium waves; mobilization; influx; ryanodine receptors; nitric oxide synthase; DAF-2; phospholipase C; astrocytes
Copyright © 2000 Society for Neuroscience 0270-6474/00/2051767-13$05.00/0
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