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A new approach to inhibiting astrocytic IP3-induced intracellular calcium increase in an astrocyte-neuron co-culture system.

机译:在星形胶质细胞-神经元共培养系统中抑制星形细胞IP3诱导的细胞内钙增加的新方法。

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摘要

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.
机译:星形胶质细胞通过肌醇1,4,5-三磷酸诱导的Ca2 +释放(IICR)依赖性机制表现出动态的Ca2 +动员,例如Ca2 +波和Ca2 +振荡。然而,人们对星形细胞Ca 2+动员的生理功能知之甚少。为了研究这个问题,我们创建了一个编码增强了绿色荧光蛋白标签的肌醇1,4,5-三磷酸吸收蛋白的质粒,并在培养的星形胶质细胞中表达了该质粒。该蛋白的表达抑制了培养的星形胶质细胞中的IICR和Ca2 +波。通过将此方法与单细胞电穿孔技术相结合,我们能够在星形胶质细胞-神经元共培养系统中特异性抑制星形胶质细胞中的IICR。我们的方法为直接检查星形细胞Ca2 +信号传导对神经元功能的生理作用提供了有用的工具。

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