Analysis of reactive astrogliosis in mouse brain using in situ hybridization combined with immunohistochemistry

Summary: 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 quantifyin...

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Bibliographic Details
Main Authors: Ranjithmenon Muraleedharan, Diana Nardini, Ronald Raymond Waclaw, Biplab Dasgupta
Format: Article
Language:English
Published: Elsevier 2021-03-01
Series:STAR Protocols
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Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721000824
Description
Summary:Summary: 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 Muraleedharan et al. (2020).
ISSN:2666-1667