Work

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

Year: 2024

Type: article

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. • Detection of astrocytes and microglia-specific complement expression using mRNA FISH • Signal-amplified detection of Aβ plaques in the TauPS2APP mouse model of AD • Measurement of glial complement expression in relation to Aβ plaques using mRNA FISH • Workflow for whole-slide imaging and spatial gene expression analysis within a week Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103388

Authors: Sadhna Rao, Joanna Yung, Melissa Gonzalez Edick, Jesse E. Hanson

Institution: Genentech

Language: English

Topic: Alzheimer's disease research and treatments

Subfield: Physiology

Field: Medicine

Domain: Health Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500