Abstract
In this study, we highlight a role for the nitric oxide-cGMP-dependent protein kinase (NO-G-kinase) signaling pathway in glial intercellular Ca(2+) wave initiation and propagation. Addition of the NO donor molsidomine (100-500 microM) or puffing aqueous NO onto primary glial cell cultures evoked an increase in [Ca(2+)](i) in individual cells and also local intercellular Ca(2+) waves, which persisted after removal of extracellular Ca(2+). High concentrations of ryanodine (100-200 microM) and antagonists of the NO-G-kinase signaling pathway essentially abrogated the NO-induced increase in [Ca(2+)](i), indicating that NO mobilizes Ca(2+) from a ryanodine receptor-linked store, via the NO-G-kinase signaling pathway. Addition of 10 microM nicardipine to cells resulted in a slowing of the molsidomine-induced rise in [Ca(2+)](i), and inhibition of Mn(2+) quench of cytosolic fura-2 fluorescence mediated by a bolus application of 2 microM aqueous NO to cells, indicating that NO also induces Ca(2+) influx in glia. Mechanical stress of individual glial cells resulted in an increase in intracellular NO in target and neighboring cells and intercellular Ca(2+) 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 Delta[Ca(2+)](i) and the rate of Ca(2+) wave propagation in these cultures. Results from this study suggest that NO-G-kinase signaling is coupled to Ca(2+) mobilization and influx in glial cells and that this pathway plays a fundamental role in the generation and propagation of intercellular Ca(2+) waves in glia.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aminoquinolines / pharmacology
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Animals
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Antineoplastic Agents / pharmacology
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Apyrase / pharmacology
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Astrocytes / chemistry
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Astrocytes / cytology
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Astrocytes / enzymology*
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Caenorhabditis elegans Proteins
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Calcium / metabolism*
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Calcium Channel Blockers / pharmacology
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Cells, Cultured
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Chelating Agents / pharmacology
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Cyclic GMP / analogs & derivatives
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Cyclic GMP / pharmacology
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Cyclic N-Oxides / pharmacology
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Egtazic Acid / pharmacology
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Enzyme Inhibitors / pharmacology
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Estrenes / pharmacology
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Free Radical Scavengers / pharmacology
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GTP-Binding Proteins / metabolism*
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Imidazoles / pharmacology
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Ionomycin / pharmacology
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Ionophores / pharmacology
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Neurons / cytology
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Nicardipine / pharmacology
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Nitric Oxide / metabolism*
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Nitric Oxide Synthase / metabolism
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Phosphodiesterase Inhibitors / pharmacology
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Potassium Chloride / pharmacology
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Prosencephalon / cytology
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Pyrrolidinones / pharmacology
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Rats
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Receptor, Insulin / metabolism
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Ryanodine / pharmacology
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Ryanodine Receptor Calcium Release Channel / physiology
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Signal Transduction / drug effects
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Signal Transduction / physiology*
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Suramin / pharmacology
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Thionucleotides / pharmacology
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Type C Phospholipases / metabolism
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omega-N-Methylarginine / pharmacology
Substances
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8-(4-chlorophenylthio)guanosine 3',5'-cyclic monophosphorothioate
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Aminoquinolines
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Antineoplastic Agents
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Caenorhabditis elegans Proteins
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Calcium Channel Blockers
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Chelating Agents
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Cyclic N-Oxides
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Enzyme Inhibitors
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Estrenes
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Free Radical Scavengers
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Imidazoles
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Ionophores
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Phosphodiesterase Inhibitors
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Pyrrolidinones
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Ryanodine Receptor Calcium Release Channel
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Thionucleotides
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1-(6-((3-methoxyestra-1,3,5(10)-trien-17-yl)amino)hexyl)-1H-pyrrole-2,5-dione
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Ryanodine
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2-phenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide
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omega-N-Methylarginine
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Nitric Oxide
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Egtazic Acid
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Ionomycin
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Suramin
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Potassium Chloride
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6-anilino-5,8-quinolinedione
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Nicardipine
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Nitric Oxide Synthase
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DAF-2 protein, C elegans
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Receptor, Insulin
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Type C Phospholipases
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GTP-Binding Proteins
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Apyrase
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Cyclic GMP
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Calcium