Sex-dependent polymorphism in urinary output is reversed upon deletion of exchange protein directly activated by cAMP (Epac)

It is well-described that there are notable sex-dependent differences in urinary production with females having a smaller volume and greater urinary concentrating ability. Apart from contribution of the androgenic inputs, the underlying signaling intermediates remain largely unexplored. We previously showed that exchange protein directly activated by cAMP (Epac) is a critical regulator of water-solute transport in the proximal tubule and collecting duct. Deletion of either Epac1 or Epac2 isoforms increases urinary output and Na + excretion. Here, we used metabolic cage balance studies to test whether Epac signaling mediates sex-dependent differences in urinary production. We found that concomitant deletion of Epac1&2 moderately increases 24h urinary volume by 38% (from 1.82+/-0.13 to 2.52+/-0.21 ml) in males, but drastically augment urinary production in females by 320% (from 1.39+/-0.10 to 4.45+/-0.37 ml). This was associated with only a mild though significant reduction in urinary osmolarity in males by 35%, whereas urine osmolarity was markedly decreased from 2511+/-5 to 977+/-60 mOsm in females. Consistently, plasma osmolarity was not different in EpacWT versus Epac1&2−/− males, but became significantly more hypertonic in Epac1&2−/− females. We further determine the functional consequences of impaired Epac cascade on fluid reabsorption in the proximal tubule and collecting duct in males and females. Deletion of Epac1 and Epac2 decreases NHE-3 expression in the proximal tubule in an additive manner. In addition, Na + /H + -dependent recovery after acidification was drastically reduced in freshly isolated split-opened proximal tubules from Epac1&2−/− mice when compared to EpacWT. Interestingly, the reduction of NHE-3 expression was more pronounced in Epac1&2−/− females (by 89%) than in males (by 59%). Immunofluorescence confocal images revealed much more diffuse intracellular distribution of AQP2 water channel in Epac1&2−/− females when compared to males. Furthermore, osmotically-driven water influx was reduced in principal cells within split-opened collecting ducts of Epac1&2−/− females but not males, when compared to respective EpacWT. In summary, our results reveal a substantially greater role of Epac signaling in renal water handling in females than in males. Interestingly, Epac1&2 deletion leads to a reversed pattern of sexual dimorphism in urinary production with females excreting larger volumes of hypotonic urine than males. This research was supported by NIH-NIDDK DK117865, DK119170, AHA EIA35260097 (to O. Pochynyuk). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.