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首页> 外文期刊>Journal of Neuroscience Methods >Transgenic mice expressing a cameleon fluorescent Ca2+ indicator in astrocytes and Schwann cells allow study of glial cell Ca2+ signals in situ and in vivo.
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Transgenic mice expressing a cameleon fluorescent Ca2+ indicator in astrocytes and Schwann cells allow study of glial cell Ca2+ signals in situ and in vivo.

机译：在星形胶质细胞和雪旺氏细胞中表达喀麦隆荧光Ca2 +指示剂的转基因小鼠允许在原位和体内研究神经胶质细胞Ca2 +信号。

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摘要

Glial cell Ca2+ signals play a key role in glial-neuronal and glial-glial network communication. Numerous studies have thus far utilized cell-permeant and injected Ca2+ indicator dyes to investigate glial Ca2+ signals in vitro and in situ. Genetically encoded fluorescent Ca2+ indicators have emerged as novel probes for investigating cellular Ca2+ signals. We have expressed one such indicator protein, the YC 3.60 cameleon, under the control of the S100beta promoter and directed its expression predominantly in astrocytes and Schwann cells. Expression of YC 3.60 extended into the entire cellular cytoplasmic compartment and the fine terminal processes of protoplasmic astrocytes and Schwann cell Cajal bands. In the brain, all the cells known to express S100beta in the adult or during development, expressed YC 3.60. While expression was most extensive in astrocytes, other glial cell types that express S100beta, such as NG2 and CNP-positive oligodendrocyte progenitor cells (OP cells), microglia, and some of the large motor neurons in the brain stem, also contained YC 3.60 fluorescence. Using a variety of known in situ and in vivo assays, we found that stimuli known to elicit Ca2+ signals in astrocytes caused substantial and rapid Ca2+ signals in the YC 3.60-expressing astrocytes. In addition, forepaw stimulation while imaging astrocytes through a cranial window in the somatosensory cortex in live mice, revealed robust evoked and spontaneous Ca2+ signals. These results, for the first time, show that genetically encoded reporter is capable of recording activity-dependent Ca2+ signals in the astrocyte processes, and networks.
机译：神经胶质细胞Ca 2+信号在神经胶质-神经元和神经胶质-神经胶质网络通信中起关键作用。迄今为止，许多研究已经利用细胞渗透性和注入的Ca2 +指示剂染料在体外和原位研究神经胶质Ca2 +信号。遗传编码的荧光Ca2 +指示剂已成为研究细胞Ca2 +信号的新型探针。我们已经在S100beta启动子的控制下表达了一种这样的指示蛋白YC 3.60 cameleon，并指导其在星形胶质细胞和雪旺氏细胞中的表达。 YC 3.60的表达延伸到整个细胞质室以及原生质星形胶质细胞和雪旺氏细胞Cajal带的精细末端过程。在大脑中，所有已知在成年或发育过程中表达S100beta的细胞均表达YC 3.60。虽然星形胶质细胞中表达最广泛，但其他表达S100beta的神经胶质细胞类型（例如NG2和CNP阳性少突胶质祖细胞（OP细胞），小胶质细胞和脑干中的一些大型运动神经元）也包含YC 3.60荧光。使用各种已知的原位和体内试验，我们发现已知在星形胶质细胞中引起Ca2 +信号的刺激在表达YC 3.60的星形胶质细胞中引起大量且快速的Ca2 +信号。此外，在活体小鼠的体感皮层中通过颅窗对星形胶质细胞成像时，前爪刺激显示出强大的诱发和自发的Ca2 +信号。这些结果首次表明，遗传编码的报告基因能够在星形胶质细胞的过程和网络中记录依赖活性的Ca2 +信号。

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