Transmembrane Serine Protease 2 and Proteolytic Activation of the Epithelial Sodium Channel in Mouse Kidney

Sure F, Afonso S, Essigke D, Schmidt P, Kalo MZ, Nesterov V, Kißler A, Bertog M, Rinke R, Wittmann S, Broeker KA, Gramberg T, Artunc F, Korbmacher C, Ilyaskin A (2024)

Publication Type: Journal article

Publication year: 2024

Journal

Article Number: 10.1681/ASN.0000000521

DOI: 10.1681/ASN.0000000521

Abstract

Background The renal epithelial sodium channel (ENaC) is essential for sodium balance and blood pressure control. ENaC undergoes complex proteolytic activation by not yet clearly identified tubular proteases. Here, we examined a potential role of transmembrane serine protease 2 (TMPRSS2). Methods Murine ENaC and TMPRSS2 were (co-)expressed in Xenopus laevis oocytes. ENaC cleavage and function were studied in TMPRSS2-deficient murine cortical collecting duct (mCCDcl1) cells and TMPRSS2-knockout (Tmprss2^−/−) mice. Short-circuit currents (ISC) were measured to assess ENaC-mediated transepithelial sodium transport of mCCDcl1 cells. The mCCDcl1 cell transcriptome was studied using RNA sequencing. The effect of low-sodium diet with or without high potassium were compared in Tmprss2^−/− and wildtype mice using metabolic cages. ENaC-mediated whole-cell currents were recorded from microdissected tubules of Tmprss2^−/− and wildtype mice. Results In oocytes, co-expression of murine TMPRSS2 and ENaC resulted in fully cleaved γ-ENaC and ~2-fold stimulation of ENaC currents. High baseline expression of TMPRSS2 was detected in mCCDcl1 cells without a stimulatory effect of aldosterone on its function or transcription. TMPRSS2 knockout in mCCDcl1 cells compromised γ-ENaC cleavage and reduced baseline and aldosterone-stimulated ISC which could be rescued by chymotrypsin. A compensatory transcriptional upregulation of other proteases was not observed. Tmprss2^−/− mice kept on standard diet exhibited no apparent phenotype, but renal γ-ENaC cleavage was altered. In response to a low-salt diet, particularly with high potassium intake, Tmprss2^−/− mice increased plasma aldosterone significantly more than wildtype mice to achieve a similar reduction of renal sodium excretion. Importantly, the stimulatory effect of trypsin on renal tubular ENaC currents was much more pronounced in Tmprss2^−/− mice than that in wildtype mice. This indicated the presence of incompletely cleaved and less active channels at the cell surface of TMPRSS2-deficient tubular epithelial cells. Conclusions TMPRSS2 contributes to proteolytic ENaC activation in mouse kidney in vivo.

Authors with CRIS profile

Involved external institutions

Universitätsklinikum Tübingen Germany (DE) Universität Regensburg Germany (DE)

How to cite

APA:

Sure, F., Afonso, S., Essigke, D., Schmidt, P., Kalo, M.Z., Nesterov, V.,... Ilyaskin, A. (2024). Transmembrane Serine Protease 2 and Proteolytic Activation of the Epithelial Sodium Channel in Mouse Kidney. Journal of the American Society of Nephrology. https://doi.org/10.1681/ASN.0000000521

MLA:

Sure, Florian, et al. "Transmembrane Serine Protease 2 and Proteolytic Activation of the Epithelial Sodium Channel in Mouse Kidney." Journal of the American Society of Nephrology (2024).

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