In Situ Patch-seq Analysis of Microglia Reveals a Lack of Stress Genes As Found in FACS-isolated Microglia

Overview

Journal PLoS One

Specialties General Medicine
Science

Date 2024 Jul 11

PMID 38990806

Authors

Olga Bakina

Thomas Conrad

Nina Mitic

Ramon Oliveira Vidal

Tessa Obrusnik

Somesh Sai

Christiane Nolte

Marcus Semtner

Helmut Kettenmann

Affiliations

Soon will be listed here.

Abstract

We applied the patch-seq technique to harvest transcripts from individual microglial cells from cortex, hippocampus and corpus callosum of acute brain slices from adult mice. After recording membrane currents with the patch-clamp technique, the cytoplasm was collected via the pipette and underwent adapted SMART-seq2 preparation with subsequent sequencing. On average, 4138 genes were detected in 113 cells from hippocampus, corpus callosum and cortex, including microglia markers such as Tmem119, P2ry12 and Siglec-H. Comparing our dataset to previously published single cell mRNA sequencing data from FACS-isolated microglia indicated that two clusters of cells were absent in our patch-seq dataset. Pathway analysis of marker genes in FACS-specific clusters revealed association with microglial activation and stress response. This indicates that under normal conditions microglia in situ lack transcripts associated with a stress-response, and that the microglia-isolation procedure by mechanical dissociation and FACS triggers the expression of genes related to activation and stress.

Citing Articles

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