Single-cell analysis of mesenchymal cells in permeable neural vasculature reveals novel diverse subpopulations of fibroblasts

Abstract

Background: In the choroid plexus and pituitary gland, vasculature is known to have a permeable, fenestrated phenotype which allows for the free passage of molecules in contrast to the blood brain barrier observed in the rest of the CNS. The endothelium of these compartments, along with secretory, neural-lineage cells (choroid epithelium and pituitary endocrine cells) have been studied in detail, but less attention has been given to the perivascular mesenchymal cells of these compartments.

Methods: The Hic1^CreERT2 Rosa26^LSL-TdTomato mouse model was used in conjunction with a Pdgfra^H2B-EGFP mouse model to examine mesenchymal cells, which can be subdivided into Pdgfra⁺ fibroblasts and Pdgfra^- pericytes within the choroid plexus (CP) and pituitary gland (PG), by histological, immunofluorescence staining and single-cell RNA-sequencing analyses.

Results: We found that both CP and PG possess substantial populations of distinct Hic1⁺ mesenchymal cells, including an abundance of Pdgfra⁺ fibroblasts. Within the pituitary, we identified distinct subpopulations of Hic1⁺ fibroblasts in the glandular anterior pituitary and the neurosecretory posterior pituitary. We also identified multiple distinct markers of CP, PG, and the meningeal mesenchymal compartment, including alkaline phosphatase, indole-n-methyltransferase and CD34.

Conclusions: Novel, distinct subpopulations of mesenchymal cells can be found in permeable vascular interfaces, including the CP, PG, and meninges, and make distinct contributions to both organs through the production of structural proteins, enzymes, transporters, and trophic molecules.

Keywords: Hic1; Blood brain barrier; Choroid plexus; Fibroblasts; Meninges; Mesenchymal cells; Neurovascular; Pericytes; Pituitary; scRNA-seq.