Abstract
Astrocytes exhibit dynamic Ca2+ mobilization, such as Ca2+ wave and Ca2+ oscillation, via an inositol 1,4,5-triphosphate-induced Ca2+ release (IICR)-dependent mechanism. The physiological functions of astrocytic Ca2+ mobilization, however, are poorly understood. To investigate this issue, we created a plasmid encoding an enhanced green fluorescent protein-tagged inositol 1,4,5-triphosphate absorbent protein and expressed it in cultured astrocytes. Expression of this protein inhibited both IICR and the Ca2+ wave in cultured astrocytes. By combining this method to the single cell electroporation technique, we were able to inhibit IICR specifically in astrocytes in an astrocyte-neuron co-culture system. Our approach provides a useful tool for direct examination of the physiological role of astrocytic Ca2+ signaling on neuronal function.
Publication types
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Comparative Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Adenosine Triphosphate / pharmacology
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Animals
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Astrocytes / drug effects*
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Astrocytes / metabolism
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Brain / cytology
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Calcium / metabolism*
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Calcium Channels / genetics
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Calcium Channels / metabolism
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Calcium Signaling / drug effects*
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Cell Communication
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Cells, Cultured
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Coculture Techniques / methods
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Drug Interactions
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Electroporation / methods
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Enzyme Inhibitors / pharmacology
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Extracellular Space / drug effects
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Fura-2 / metabolism
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Glycoproteins
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Green Fluorescent Proteins / biosynthesis
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Inositol 1,4,5-Trisphosphate / pharmacology*
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Inositol 1,4,5-Trisphosphate Receptors
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Mice
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Mutation
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Neurons / physiology*
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Potassium Chloride / pharmacology
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Receptors, Cytoplasmic and Nuclear / genetics
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Receptors, Cytoplasmic and Nuclear / metabolism
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Thapsigargin / pharmacology
Substances
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Calcium Channels
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Enzyme Inhibitors
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Glycoproteins
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Inositol 1,4,5-Trisphosphate Receptors
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Receptors, Cytoplasmic and Nuclear
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Green Fluorescent Proteins
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Potassium Chloride
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Thapsigargin
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Inositol 1,4,5-Trisphosphate
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Adenosine Triphosphate
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Calcium
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Fura-2