Altered Subcellular Distribution of Na+,K+-ATPase in Proximal Tubules in Young Spontaneously Hypertensive Rats

During early development of hypertension, the spontaneously hypertensive rat (SHR) demonstrates increased proximal tubule sodium reabsorption. Our previous observations of reduced Na + ,K + -ATPase catalytic α1 and γ subunit transcript abundance in SHR proximal tubule led us to test the hypothesis that increased proximal tubule sodium reabsorption may be attributable to altered subunit protein abundance, post-translational modification, or a shift in subcellular α1 and γ distribution toward the basolateral membrane. We now extend previous gene expression studies by analyzing total cellular α1 and γ protein abundance in proximal tubule from SHR compared with matched Wistar–Kyoto (WKY) controls. We also used sucrose density-gradient centrifugation to isolate basolateral, early, and late endosomal membrane–enriched fractions as well as cell surface biotinylation to test the hypothesis of altered subunit subcellular distribution in the SHR proximal tubule. At 4 weeks of age, significantly greater amounts of α1 were present in basolateral membrane–enriched fractions of SHR than WKY (21.1±1.8% versus 12.3±1.8%; P P P P + ,K + -ATPase subunits.