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联合荧光原位杂交（FISH）和免疫组化（IHC）检测小鼠大脑中与淀粉样斑块相关的胶质细胞补体表达的方案。

Protocol for detection of glial complement expression in relation to amyloid plaques in mouse brain with combined FISH and IHC.

作者信息

Rao Sadhna, Yung Joanna, Edick Melissa Gonzalez, Hanson Jesse E

机构信息

Genentech, Inc., South San Francisco, CA 94080, USA.

Genentech, Inc., South San Francisco, CA 94080, USA.

出版信息

STAR Protoc. 2024 Dec 20;5(4):103388. doi: 10.1016/j.xpro.2024.103388. Epub 2024 Oct 16.

DOI:10.1016/j.xpro.2024.103388
PMID:39418161
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11530921/
Abstract

Complement proteins contribute to neurodegeneration in Alzheimer's disease (AD) and are secreted by glia surrounding beta-amyloid (Aβ) plaques. We present an optimized protocol for Aβ plaque detection with tyramide-digoxigenin signal amplification. This is combined with a multiplex mRNA fluorescence in situ hybridization (FISH) panel to assay glial-specific complement expression proximal to Aβ plaques in TauPS2APP mice. We describe steps for tissue preparation and mRNA detection. We then detail steps for the detection of Aβ plaques, image acquisition, and analysis.

摘要

补体蛋白在阿尔茨海默病（AD）中促成神经退行性变，且由围绕β-淀粉样蛋白（Aβ）斑块的神经胶质细胞分泌。我们提出了一种用酪胺-地高辛信号放大法检测Aβ斑块的优化方案。该方案与多重mRNA荧光原位杂交（FISH）检测板相结合，以检测TauPS2APP小鼠中Aβ斑块附近神经胶质细胞特异性补体的表达。我们描述了组织制备和mRNA检测的步骤。然后详细介绍了Aβ斑块检测、图像采集和分析的步骤。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/71c15e2f7129/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/b4fefb8e1146/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/a37c9d2bafdc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/4291dd07aac6/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/1fdbfa857817/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/71c15e2f7129/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/b4fefb8e1146/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/a37c9d2bafdc/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/4291dd07aac6/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/1fdbfa857817/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/49d6/11530921/71c15e2f7129/gr4.jpg

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本文引用的文献

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