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

Reactive astrogliosis is characterized by a profound change in astrocyte pheno-type in response to all CNS injuries. Here, we present a revised in situ hybridiza-tion 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 hybridiza-tion followed by IHC. We provide a staining protocol for quantitative measures of astrocyte reactivity as an independent confirmation of the magnitude of reac-tive gliosis. For complete details on the use and execution of this protocol, please refer to Muraleedharan et al. (2020).

Automated summary

Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to CNS injuries. This study presents a revised protocol for labeling reactive astrocytes in mouse brains and provides a staining protocol for quantitative measures of astrocyte reactivity.