Astrocytes are a major type of glial cell in the mammalian brain, essentially regulating neuronal development and function. Quantitative imaging represents an important approach to study astrocytic signaling in neural circuits. Focusing on astrocytic Ca2+ activity, a key pathway implicated in astrocye-neuron interaction, we here report a strategy combining fast light sheet fluorescence microscopy (LSFM) and correlative screening-based time series analysis, to map activity domains in astrocytes in living mammalian nerve tissue. Light sheet of micron-scale thickness enables wide-field optical sectioning to image astrocytes in acute mouse brain slices. Using both chemical and genetically encoded Ca2+ indicators, we demonstrate the complementary advantages of LSFM in mapping Ca2+ domains in astrocyte populations as compared to epifluorescence and two-photon microscopy. Our approach then revealed distinct kinetics of Ca2+ signals in cortical and hypothalamic astrocytes in resting conditions and following the activation of adrenergic G protein coupled receptor (GPCR). This observation highlights the activity heterogeneity across regionally distinct astrocyte populations, and indicates the potential of our method for investigating dynamic signals in astrocytes.

中文翻译：

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通过光片成像和时空相关筛选绘制星形胶质细胞活动域

星形胶质细胞是哺乳动物大脑中一种主要类型的神经胶质细胞，主要调节神经元的发育和功能。定量成像是研究神经回路中星形胶质细胞信号传导的重要方法。专注于星形胶质细胞 Ca2+ 活性，这是一种与星形细胞-神经元相互作用有关的关键途径，我们在这里报告了一种结合快速光片荧光显微镜 (LSFM) 和基于相关筛选的时间序列分析的策略，以绘制活体哺乳动物神经组织中星形胶质细胞的活性域. 微米级厚度的光片使宽视野光学切片能够对急性小鼠脑切片中的星形胶质细胞进行成像。使用化学和遗传编码的 Ca2+ 指示剂，与落射荧光和双光子显微镜相比，我们证明了 LSFM 在映射星形胶质细胞群中 Ca2+ 结构域方面的互补优势。我们的方法随后揭示了皮层和下丘脑星形胶质细胞在静息条件下和肾上腺素能 G 蛋白偶联受体 (GPCR) 激活后 Ca2+ 信号的不同动力学。这一观察结果突出了区域不同星形胶质细胞群体的活动异质性，并表明了我们研究星形胶质细胞动态信号的方法的潜力。