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Low Na +  intake activates aldosterone signaling, which increases renal Na +  reabsorption through increased apical activity of the NaCl cotransporter (NCC) and the epithelial Na +  channel (ENaC). Na +  transporter proteins are excreted in urine as an integral part of cell-derived extracellular vesicles (uEVs). It was hypothesized that Na +  transport protein levels in uEVs from healthy humans reflect their physiological regulation by aldosterone. Urine and plasma samples from 10 healthy men (median age: 22.8 yr) were collected after 5 days on a low-Na +  (70 mmol/day) diet and 5 days on a high-Na +  (250 mmol/day) diet. uEVs were isolated by ultracentrifugation and analyzed by Western blot analysis for EV markers (CD9, CD63, and ALIX), transport proteins (Na + -K + -ATPase α 1 -subunit, NCC, ENaC α- and γ-subunits, and aquaporin 2), and the ENaC-cleaving protease prostasin. Plasma renin and aldosterone concentrations increased during the low-Na +  diet. uEV size and concentration were not different between diets by tunable resistive pulse sensing. EV markers ALIX and CD9 increased with the low-Na +  diet, whereas CD63 and aquaporin 2 excretion were unchanged. Full-length ENaC γ-subunits were generally not detectable in uEVs, whereas ENaC α-subunits, NCC, and phosphorylated NCC were consistently detected but not changed by Na +  intake. Prostasin increased with low Na +  in uEVs. uEV excretion of transporters was not correlated with blood pressure, urinary Na +  and K +  excretion, plasma renin, or aldosterone. In conclusion, apical Na +  transporter proteins and proteases were excreted in uEVs, and while the excretion rate and size of uEVs were not affected, EV markers and prostasin increased in response to the low-Na +  diet.

american journal of physiology. renal physiology./american journal of physiology. renal physiology

Dietary Na<sup>+</sup>intake in healthy humans changes the urine extracellular vesicle prostasin abundance while the vesicle excretion rate, NCC, and ENaC are not altered

Dietary Na+intake in healthy humans changes the urine extracellular vesicle prostasin abundance while the vesicle excretion rate, NCC, and ENaC are not altered

Dietary Na⁺intake in healthy humans changes the urine extracellular vesicle prostasin abundance while the vesicle excretion rate, NCC, and ENaC are not altered