Sulforhodamine 101 as a specific marker of astroglia in the neocortex in vivo
Sulforhodamine 101 as a specific marker of astroglia in the neocortex in vivo
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DOI：
10.1038/nmeth706
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发表时间：
2004-10-01
期刊：
影响因子：
48
通讯作者：
Helmchen, F
中科院分区：
文献类型：
--
作者：
Nimmerjahn, A;Kirchhoff, F;Helmchen, F
神经胶质细胞已被确定为大脑中的关键信号成分;然而，在体内研究其结构和功能的方法一直缺乏。在这里，我们描述了一种新的，高选择性的方法标记星形胶质细胞在完整的啮齿动物新皮层，允许在体内成像使用双光子显微镜。红色荧光染料磺酰罗丹明101（SR 101），特别是采取了由原生质星形胶质细胞短暂暴露于脑表面后。通过免疫组织化学证实特异性。此外，SR 101标记了转基因小鼠中表达增强型绿色荧光蛋白（EGFP）的星形胶质细胞，但不标记小胶质细胞。我们使用SR 101标记来量化星形胶质细胞的形态学特征，并可视化它们与皮质微血管的密切联系。此外，通过将这种方法与细胞群的钙指示剂加载相结合，我们证明了星形胶质细胞和神经元网络中不同的钙动力学。我们期望SR 101染色成为研究体内星形胶质细胞的主要工具。
Glial cells have been identified as key signaling components in the brain; however, methods to investigate their structure and function in vivo have been lacking. Here, we describe a new, highly selective approach for labeling astrocytes in intact rodent neocortex that allows in vivo imaging using two-photon microscopy. The red fluorescent dye sulforhodamine 101 (SR101) was specifically taken up by protoplasmic astrocytes after brief exposure to the brain surface. Specificity was confirmed by immunohistochemistry. In addition, SR101 labeled enhanced green fluorescent protein (EGFP)-expressing astrocytes but not microglial cells in transgenic mice. We used SR101 labeling to quantify morphological characteristics of astrocytes and to visualize their close association with the cortical microvasculature. Furthermore, by combining this method with calcium indicator loading of cell populations, we demonstrated distinct calcium dynamics in astroglial and neuronal networks. We expect SR101 staining to become a principal tool for investigating astroglia in vivo.