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| Item Type: | Article |
|---|---|
| Title: | In situ Patch-seq analysis of microglia reveals a lack of stress genes as found in FACS-isolated microglia |
| Creators Name: | Bakina, O., Conrad, T., Mitic, N., Vidal, R.O., Obrusnik, T., Sai, S., Nolte, C., Semtner, M. and Kettenmann, H. |
| 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. |
| Keywords: | Flow Cytometry, Inbred C57BL Mice, Microglia, Patch-Clamp Techniques, Single-Cell Analysis, Physiological Stress, Animals, Hippocampus, Mice |
| Source: | PLoS ONE |
| ISSN: | 1932-6203 |
| Publisher: | Public Library of Science |
| Volume: | 19 |
| Number: | 7 |
| Page Range: | e0302376 |
| Date: | 11 July 2024 |
| Official Publication: | https://doi.org/10.1371/journal.pone.0302376 |
| PubMed: | View item in PubMed |
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