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Effects of long‐term aldosterone administration on mRNA expression levels in the renal distal nephron examined by FACS and RNA sequencing (1088.13)
Author(s) -
Poulsen Søren,
Praetorius Jeppe,
Fenton Robert,
Christensen Birgitte
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.1088.13
Subject(s) - aldosterone , nephron , messenger rna , distal convoluted tubule , medicine , endocrinology , rna , cell sorting , microbiology and biotechnology , biology , epithelial sodium channel , kidney , aldosterone synthase , renal sodium reabsorption , gene , chemistry , blood pressure , renin–angiotensin system , reabsorption , sodium , genetics , flow cytometry , organic chemistry
The renal distal nephron [distal convoluted tubule 2 (DCT2) and connecting tubule (CNT)] is crucial for sodium and blood pressure regulation, and is tightly controlled by aldosterone. Due to difficulties in isolating the DCT2 and CNT, only few studies have examined the regulation of cellular processes in these segments. We have generated a mouse model expressing EGFP in the DCT2 and CNT, making it possible to collect EGFP cells using FACS sorting. Combined with the emergence of next generation sequencing (RNA sequencing), we utilized these tools to generate an improved catalog of aldosterone‐regulated mRNA transcripts in the distal nephron. The EGFP mice were administrated saline (n=7) or aldosterone (n=7) for six days. Following, a population of EGFP positive cells was collected using FACS sorting (purity~90%). RNA sequencing results confirmed that mRNA transcripts classically known to be regulated by aldosterone were changed in abundance [NCC and αENaC tended to be increased (p=0.058 and 0.147, respectively), Sgk1 was increased (p=0.003), and Nedd4‐2 was decreased (p=0.011)]. Based on regular p‐values (significance level set to 0.05), 562 genes differentially regulated by aldosterone were identified. In particular, genes involved in processes in the nucleus, mitochondrion, and plasma membrane were highly represented. The results provide us with the opportunity to identify novel mRNAs involved in renal blood pressure regulation.