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

STAR Protocols

Ronald Raymond Waclaw¹  Biplab Dasgupta²  Diana Nardini²  Ranjithmenon Muraleedharan³ 

[1] Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45229, USA;Division of Experimental Hematology and Cancer Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA;Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA;

关键词: Cancer;    Metabolism;    Microscopy;    Model organisms;    In situ hybridization;    Neuroscience;   

DOI  :  

来源: DOAJ

【 摘 要 】

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).

【 授权许可】

Unknown