| First Author | Bakina O | Year | 2024 |
| Journal | PLoS One | Volume | 19 |
| Issue | 7 | Pages | e0302376 |
| PubMed ID | 38990806 | Mgi Jnum | J:351141 |
| Mgi Id | MGI:7664978 | Doi | 10.1371/journal.pone.0302376 |
| Citation | Bakina O, et al. (2024) In situ Patch-seq analysis of microglia reveals a lack of stress genes as found in FACS-isolated microglia. PLoS One 19(7):e0302376 |
| abstractText | 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. |
