利用杂交结合免疫组织化学分析小鼠大脑中的反应性星形胶质细胞增生。

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

机构信息

Division of Oncology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.

Division of Experimental Hematology and Cancer Biology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH 45229, USA.

出版信息

STAR Protoc. 2021 Mar 2;2(1):100375. doi: 10.1016/j.xpro.2021.100375. eCollection 2021 Mar 19.

DOI:10.1016/j.xpro.2021.100375

PMID:33718893

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7933799/

Abstract

Reactive astrogliosis is characterized by a profound change in astrocyte phenotype in response to all CNS injuries. Here, we present a revised 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 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).

摘要

反应性星形胶质细胞增生以星形胶质细胞表型的深刻变化为特征，以响应所有中枢神经系统损伤。在这里，我们提出了一种改进的杂交和免疫组织化学（IHC）方案，以标记小鼠大脑中的反应性星形胶质细胞。几种量化星形胶质细胞反应性的方法缺乏敏感性来区分整个谱。我们优化了杂交，然后是 IHC。我们提供了一种染色方案，用于定量测量星形胶质细胞反应性，作为对反应性神经胶质形成程度的独立验证。有关此方案的使用和执行的完整详细信息，请参阅 Muraleedharan 等人。（2020 年）。