A high-throughput single-cell RNA expression profiling method identifies human pericyte markers.

AIMS: We sought to identify and optimize a universally available histological marker for pericytes in the human brain. Such a marker could be a useful tool for researchers. Further, identifying a gene expressed relatively specifically in human pericytes could provide new insights into the biological functions of this fascinating cell type.

METHODS: We analysed single-cell RNA expression profiles derived from different human and mouse brain regions using a high-throughput and low-cost single-cell transcriptome sequencing method called EasySci. Throughthis analysis, we were able to identify specific gene markers for pericytes, some of which had not been previously characterized. We then used commercially (and therefore universally) available antibodies to immunolabel the pericyte-specific gene products in formalin-fixed paraffin-embedded (FFPE) human brains, and also performed immunoblots to determine whether appropriately sized proteins were recognized.

RESULTS: In the EasySci data sets, highly pericyte-enriched expression was notable for SLC6A12 and SLC19A1. Antibodies against these proteins recognized bands of the correct size in immunoblots of human brain extracts. Following optimization of the immunohistochemical technique, staining for both antibodies was strongly positive in small blood vessels, and was far more effective than a PDGFRB antibody at staining pericyte-like cells in FFPE human brain sections. In an exploratory sample of other human organs (kidney, lung, liver, muscle), immunohistochemistry did not show the same pericyte-like pattern of staining.

CONCLUSIONS: The SLC6A12 antibody was well suited for labelling pericytes in human FFPE brain sections, based on the combined results of single-cell RNA-seq analyses, immunoblots, and immunohistochemical studies.