Protein phosphatase 1 inhibitor-1 deficiency reduces phosphorylation of renal NaCl cotransporter and causes arterial hypotension

Nicolas Picard, Katja Trompf, Chao-Ling Yang, R. Lance Miller, Monique Carrel, Dominique Loffing-Cueni, Robert A. Fenton, David Ellison, Johannes Loffing

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

The thiazide-sensitive NaCl cotransporter (NCC) of the renal distal convoluted tubule (DCT) controls ion homeostasis and arterial BP. Loss-of-function mutations of NCC cause renal salt wasting with arterial hypotension (Gitelman syndrome). Conversely, mutations in the NCC-regulating WNK kinases or kelch-like 3 protein cause familial hyperkalemic hypertension. Here, we performed automated sorting of mouse DCTs and microarray analysis for comprehensive identification of novel DCT-enriched gene products, which may potentially regulate DCT and NCC function. This approach identified protein phosphatase 1 inhibitor-1 (I-1) as a DCT-enriched transcript, and immunohistochemistry revealed I-1 expression in mouse and human DCTs and thick ascending limbs. In heterologous expression systems, coexpression of NCC with I-1 increased thiazide-dependent Na+ uptake, whereas RNAi-mediated knockdown of endogenous I-1 reduced NCC phosphorylation. Likewise, levels of phosphorylated NCC decreased by approximately 50% in I-1 (I-1-/-) knockout mice without changes in total NCC expression. The abundance and phosphorylation of other renal sodium-transporting proteins, including NaPi-IIa, NKCC2, and ENaC, did not change, although the abundance of pendrin increased in these mice. The abundance, phosphorylation, and subcellular localization of SPAK were similar in wild-type (WT) and I-1-/- mice. Compared with WT mice, I-1-/- mice exhibited significantly lower arterial BP but did not display other metabolic features of NCC dysregulation. Thus, I-1 is a DCT-enriched gene product that controls arterial BP, possibly through regulation of NCC activity.

Original languageEnglish (US)
Pages (from-to)511-522
Number of pages12
JournalJournal of the American Society of Nephrology
Volume25
Issue number3
DOIs
StatePublished - Mar 2014

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Hypotension
Phosphorylation
Kidney
Gitelman Syndrome
Distal Kidney Tubule
Thiazides
Mutation
Microarray Analysis
RNA Interference
Knockout Mice
Genes
protein phosphatase inhibitor-1
Proteins
Homeostasis
Phosphotransferases
Extremities
Salts
Sodium
Immunohistochemistry
Ions

ASJC Scopus subject areas

  • Nephrology

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Protein phosphatase 1 inhibitor-1 deficiency reduces phosphorylation of renal NaCl cotransporter and causes arterial hypotension. / Picard, Nicolas; Trompf, Katja; Yang, Chao-Ling; Miller, R. Lance; Carrel, Monique; Loffing-Cueni, Dominique; Fenton, Robert A.; Ellison, David; Loffing, Johannes.

In: Journal of the American Society of Nephrology, Vol. 25, No. 3, 03.2014, p. 511-522.

Research output: Contribution to journalArticle

Picard, Nicolas ; Trompf, Katja ; Yang, Chao-Ling ; Miller, R. Lance ; Carrel, Monique ; Loffing-Cueni, Dominique ; Fenton, Robert A. ; Ellison, David ; Loffing, Johannes. / Protein phosphatase 1 inhibitor-1 deficiency reduces phosphorylation of renal NaCl cotransporter and causes arterial hypotension. In: Journal of the American Society of Nephrology. 2014 ; Vol. 25, No. 3. pp. 511-522.
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