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  • Title: Altered subcellular distribution of Na+,K+-ATPase in proximal tubules in young spontaneously hypertensive rats.
    Author: Hinojos CA, Doris PA.
    Journal: Hypertension; 2004 Jul; 44(1):95-100. PubMed ID: 15173126.
    Abstract:
    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 alpha1 and gamma 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 alpha1 and gamma distribution toward the basolateral membrane. We now extend previous gene expression studies by analyzing total cellular alpha1 and gamma 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 alpha1 were present in basolateral membrane-enriched fractions of SHR than WKY (21.1+/-1.8% versus 12.3+/-1.8%; P<0.005), and there was a concomitant reduction of alpha1 in late endosomal membrane-enriched fractions of SHR (63.3+/-2.7% versus 74.8+/-4.3%; P<0.05). This finding was confirmed in cell surface biotinylation studies that showed higher alpha1 (1.45+/-0.1-fold greater; P<0.05) and gamma-subunit (3.48+/-0.7-fold greater; P<0.01) abundance in 4-week-old SHR proximal tubule plasma membrane compared with matched WKY samples. These studies support the hypothesis that development of hypertension in SHR may involve an altered subcellular distribution of proximal tubule Na+,K+-ATPase subunits.
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