back to Hyun Jun Jung's Database

RNA-Seq in Mouse Kidney ASDN with MR (NR3C2) Knockout

RNA sequencing (RNA-seq)-based transcriptomic profiling was performed in aldosterone-sensitive distal nephron (ASDN), including connecting tubule and cortical collecting duct tubule, microdissected from kidneys of MR (Nr3c2) knockout (KO) mice.

Nr3c2-flox/Pax8‐rtTA/LC1 mice were used as a doxycycline (DOX)-inducible Nr3c2 gene KO model. For these studies, four treatment groups were examined, comparing control and MR-KO mice fed different diets (Control, Normal Potassium, NK) or that either stimulate aldosterone (High Potassium Diet, HK) or limit hyperkalemia (Low Potassium Diet, LK), a significant pathophysiological sequela of MR-KO. RNA-Seq analysis was carried out in the microdissected connecting tubule and cortical collecting duct tubule segments (5-6 mice per group; ~10 fresh ASDN tubules per mouse). [Potassium diet] Normal K, 1% KCl; High K, 2.1% KCl; Low K, <0.003% KCl. [MR/GR binding] Tier1, -1kb stream to +100bp of TSS; Tier2, -3kb stream to +1kb of TSS; Tier3, -5kb upstream of TSS and gene body. FDR(False discovery rate, BH-corrected) < 0.05 is considered as significantly changed genes. As expected, the transcriptomes of control mice on a HK diet, which stimulates aldosterone secretion, and of MR-KO mice on a LK rescue diet, which protects against hyperkalemia, were most different and had the most significant number of differentially expressed genes. The differentially expressed genes between these groups comprise the MR-regulated gene atlas, as they best define aldosterone-MR action without confounding effects of hyperkalemia.

Raw sequence data available at NCBI GEO GSE269263. Please contact to Hyun Jun Jung (hynjn.jung@gmail.com) or Paul A. Welling (pwelling1@jhmi.edu, @Welling Lab).