Sodium absorption in the aldosterone-sensitive distal nephron is regulated by both steroid and peptide hormones. These stimulate sodium transport via the epithelial sodium channel (ENaC) by increasing both channel activity and number of channels at the apical surface [1]. A PI3-kinase-mTORC2-SGK1 pathway has been implicated in both the rapid effects of the peptide hormone insulin, as well as the delayed responses elicited by steroid hormones [2]. The aim of this study was to compare the mechanisms by which these hormones stimulate ENaC-mediated Na+ absorption. Na+ transport via ENaC was measured by recording the amiloride-sensitive current (Iami) across mpkCCDcl4 cortical collecting duct cells mounted in Ussing chambers. Individual components of the signalling pathway of interest were assessed by monitoring phosphorylation of downstream target proteins using Western blot analysis of whole cell lysates. qRT-PCR was used to monitor SGK1 mRNA abundance. Values are means ± S.E.M., compared by unpaired t-test. Dexamethasone (dex) stimulated Iami from baseline (∆Iami) by 19.6±2.6 µA∙cm-2 after 3h compared to control 1.5±1.9 µA∙cm-2 (n=7, p<0.001). This was associated with a 42.7±8.8 fold increase in SGK1 mRNA abundance (n=6, p<0.001). Phosphorylation a of downstream target of SGK1, NDRG1-Thr346,356,366, increased by 4.1±0.4 fold compared to control (n=10, p<0.01). However, phosphorylation of Akt‑Thr308 (n=9) and Akt‑Ser473 (n=5) remained unchanged following exposure to dex, indicative that PI3K/mTORC2 activity was unaltered. An inhibitor of protein translation cyclohexamide (CHX) reduced the effect of dex on Iami by 48.5±22.4 % (n=7), without altering the associated increase in SGK1 mRNA, 41.9±12.5 fold (n=6). After 1h exposure to insulin, ∆Iami was stimulated by 6.8±1.7 µA∙cm‑2 compared to control 0.2±0.7 µA∙cm-2 (n=6, p<0.01). SGK1 activity was increased indicated by a 1.7±0.1 fold increase in NDRG1-Thr346,356,366 phosphorylation (n=11, p<0.001). PI3K/mTORC2 activity was also increased with a 1.8±0.2 fold increase in Akt-Thr308 (n=17, p<0.01) and 1.7±0.1 fold increase in Akt-Ser473 (n=16, p<0.001) phosphorylation, respectively. CHX did not alter the stimulation of Iami by insulin (n=6), however CHX reduced phosphorylation of all target proteins monitored, indicating reduced PI3K/mTORC2/SGK1 activity. Together these data indicate that dexamethasone stimulates ENaC-mediated Na+ transport by increasing transcription of SGK1 without increasing PI3K/mTORC2 activity. Insulin on the other hand increases PI3K/mTORC2/SGK1 activity, but does not evoke transcription of SGK1. Insulin can however still stimulate ENaC-mediated Na+ transport with significantly reduced activity of all components of this signalling pathway, suggesting either another signalling molecule is involved or that only a low level activity of this cascade is required to permit a response.