Defects in nephrogenesis result in an expansion of the Foxd1+ stromal progenitor population
Reciprocal signaling interactions coordinate multiple aspects of kidney development. While signals from the stroma have been shown to regulate nephron progenitor cell (NPC) differentiation, much less is known about regulation of the stromal progenitor population. We sought to further examine how defects in the nephron lineage may non-autonomously affect stromal development. To aid in this analysis, we generated single nuclei RNA-seq using 10x genomics from paired E15.5 control kidneys (cre-negative and Six2creTGC) and mutant kidneys (Six2cre;Wt1c/c) to identify additional markers and gene expression changes of the Foxd1+ stromal progenitor population at this timepoint, and validated the expression of early B cell factor 3 (Ebf3), aldehyde oxidase 3 (Aox3), fibroblast activation protein (Fap), insulin growth factor 1 (Igf1), and glutamate ionotropic receptor AMPA type subunit 1 (Gria1) in control and mutant kidneys. Overall design: Kidneys were harvested from E15.5 mouse embryos by timed pregnant matings. Nuclei isolation was performed for 10x genomics single nuclei RNA-seq, targeting 5000 nuclei with 50K reads/nuclei from Six2creTGC and mutant samples and 5000 nuclei with 90K reads/nuclei from one cre-negative and mutant sample.