Experimental
nephrotic syndrome leads to activation of the
epithelial sodium channel (ENaC) by proteolysis and promotes renal
sodium retention. The membrane-anchored
serine protease prostasin (
CAP1/PRSS8) is expressed in the distal nephron and participates in proteolytic ENaC regulation by serving as a scaffold for other
serine proteases. However, it is unknown whether
prostasin is also involved in ENaC-mediated
sodium retention of experimental
nephrotic syndrome. In this study, we used genetically modified knock-in mice with Prss8 mutations abolishing its proteolytic activity (Prss8-S238A) or
prostasin activation (Prss8-R44Q) to investigate the development of
sodium retention in
doxorubicin-induced
nephrotic syndrome. Healthy Prss8-S238A and Prss8-R44Q mice had normal ENaC activity as reflected by the natriuretic response to the ENaC blocker
triamterene. After
doxorubicin injection, all genotypes developed similar
proteinuria. In all genotypes, urinary
prostasin excretion increased while renal expression was not altered. In nephrotic mice of all genotypes,
triamterene response was similarly increased, consistent with ENaC activation. As a consequence, urinary
sodium excretion dropped in all genotypes and mice similarly gained
body weight by + 25 ± 3% in Prss8-wt, + 20 ± 2% in Prss8-S238A and + 28 ± 3% in Prss8-R44Q mice (p = 0.16). In Western blots, expression of fully cleaved α- and γ-ENaC was similarly increased in nephrotic mice of all genotypes. In conclusion, proteolytic ENaC activation and
sodium retention in experimental
nephrotic syndrome are independent of the activation of
prostasin and its enzymatic activity and are consistent with the action of aberrantly filtered
serine proteases or proteasuria.