We performed single cell and bulk RNA-seq analyses on pdgfrb-positive cells to identify candidate pericyte-expressed genes. For bulk RNA-seq, we used wild type pdgfrb-positive samples from Series GSE152759 (GEO ID: GSM4625924, GSM4625925, GSM4625926) for comparison to mutant pdgfrb-positive cells in this Series. These cells had originally been isolated in parallel.
Shih YH, Portman D, Idrizi F, Grosse A et al. Integrated molecular analysis identifies a conserved pericyte gene signature in zebrafish. Development 2021 Dec 1;148(23). PMID: 34751773
Abstract
Pericytes reside in capillary beds where they share a basement membrane with endothelial cells and regulate their function. However, little is known about embryonic pericyte development, in part, due to lack of specific molecular markers and genetic tools. Here, we applied single cell RNA-sequencing (scRNA-seq) of platelet derived growth factor beta (pdgfrb)-positive cells to molecularly characterize pericytes in zebrafish larvae. scRNA-seq revealed zebrafish cells expressing mouse pericyte gene orthologs, and comparison with bulk RNA-seq from wild-type and pdgfrb mutant larvae further refined a pericyte gene set. Subsequent integration with mouse pericyte scRNA-seq profiles revealed a core set of conserved pericyte genes. Using transgenic reporter lines, we validated pericyte expression of two genes identified in our analysis: NDUFA4 mitochondrial complex associated like 2a (ndufa412a), and potassium voltage-gated channel, lsk-related family, member 4 (kcne4). Both reporter lines exhibited pericyte expression in multiple anatomical locations, and kcne4 was also detected in a subset of vascular smooth muscle cells. Thus, our integrated molecular analysis revealed a molecular profile for zebrafish pericytes and allowed us to develop new tools to observe these cells in vivo.