Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry.
Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry.
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DOI：
10.1016/j.xpro.2021.100375
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发表时间：
2021-03-19
期刊：
影响因子：
--
通讯作者：
Dasgupta B
中科院分区：
文献类型：
--
作者：
Muraleedharan R;Nardini D;Waclaw RR;Dasgupta B
反应性星形胶质细胞增生的特点是星形胶质细胞表型因所有中枢神经系统损伤而发生深刻变化。在这里，我们提出了修订后的原位杂交和免疫组织化学 (IHC) 方案来标记小鼠大脑中的反应性星形胶质细胞。量化星形胶质细胞反应性的几种方法缺乏区分整个光谱的敏感性。我们优化了原位杂交，然后进行 IHC。我们提供了定量测量星形胶质细胞反应性的染色方案，作为反应性神经胶质增生程度的独立确认。有关该协议的使用和执行的完整详细信息，请参阅。定量测量小鼠大脑中反应性神经胶质增生 双重染色可识别响应慢性 CNS 病理而发生的反应性神经胶质增生 杂交阶段包括 LCN2 和 C3 mRNA 杂交和信号发育 免疫染色阶段包括 GFAP 蛋白的抗体检测 反应性星形胶质细胞增生的特点是星形胶质细胞表型因所有 CNS 损伤而发生深刻变化。在这里，我们提出了修订后的原位杂交和免疫组织化学 (IHC) 方案来标记小鼠大脑中的反应性星形胶质细胞。量化星形胶质细胞反应性的几种方法缺乏区分整个光谱的敏感性。我们优化了原位杂交，然后进行 IHC。我们提供了定量测量星形胶质细胞反应性的染色方案，作为反应性神经胶质增生程度的独立确认。
Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised in situ hybridization and immunohistochemistry (IHC) protocol to label the reactive astrocytes in the mouse brain. Several approaches for quantifying astrocyte reactivity lacked sensitivity to discriminate across the spectrum. We optimized in situ hybridization followed by IHC. We provide a staining protocol for quantitative measures of astrocyte reactivity as an independent confirmation of the magnitude of reactive gliosis. For complete details on the use and execution of this protocol, please refer to. Quantitative measurement of reactive gliosis in mouse brain Dual staining identifies reactive gliosis occurs in response to chronic CNS pathology Hybridization phase includes LCN2 and C3 mRNA hybridization and signal development Immunostaining phase includes antibody detection of GFAP protein Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised in situ hybridization and immunohistochemistry (IHC) protocol to label the reactive astrocytes in the mouse brain. Several approaches for quantifying astrocyte reactivity lacked sensitivity to discriminate across the spectrum. We optimized in situ hybridization followed by IHC. We provide a staining protocol for quantitative measures of astrocyte reactivity as an independent confirmation of the magnitude of reactive gliosis.
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DOI：
10.1523/jneurosci.6221-11.2012
发表时间：
2012-05-02
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Zamanian JL;Xu L;Foo LC;Nouri N;Zhou L;Giffard RG;Barres BA
通讯作者：
Barres BA
影响因子：
9.9
作者：
通讯作者：
--
影响因子：
8.8
作者：
Muraleedharan R;Gawali MV;Tiwari D;Sukumaran A;Oatman N;Anderson J;Nardini D;Bhuiyan MAN;Tkáč I;Ward AL;Kundu M;Waclaw R;Chow LM;Gross C;Rao R;Schirmeier S;Dasgupta B
通讯作者：
Dasgupta B
影响因子：
6.2
作者：
Wirenfeldt, M;Dalmau, I;Finsen, B
通讯作者：
Finsen, B
影响因子：
5.3
作者：
Lian, Hong;Litvinchuk, Alexandra;Zheng, Hui
通讯作者：
Zheng, Hui