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  • 1
    ISSN: 1432-1424
    Schlagwort(e): rental tubule transport ; medullary thick ascending limb ; intracellular voltage recording ; ADH ; K+ conductance
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Summary Cellular impalements were used in combination with standard transepithelial electrical measurements to evaluate some of the determinants of the spontaneous lumen-positive voltage,V e , which attends net Cl− absorption,J Cl net , and to assess how ADH might augment bothJ Cl met andV e in the mouse medullary thick ascending limb of Henle microperfusedin vitro. Substituting luminal 5mm Ba++ for 5mm K+ resulted in a tenfold increase in the apical-to-basal membrane resistance ratio,R c /R bl , and increasing luminal K+ from 5 to 50mm in the presence of luminal 10−4 m furosemide resulted in a 53-mV depolarization of apical membrane voltage,V a . Thus K+ accounted for at least 85% of apical membrane conductance. Either with or without ADH. 10−4 m luminal furosemide reducedV e andJ Cl net to near zero values and hyperpolarized bothV a andV bl , the voltage across basolateral membranes; however, the depolarization ofV bl was greater in the presence than in the absence of hormone while the hormone had no significant effect on the depolarization ofV a , Thus ADH-dependent increases inV b were referable to greater depolarizations ofV bl in the presence of ADH than in the absence of ADH 68% of the furosemide-induced hyperpolarization ofV a was referable to a decrease in the K+ current across apical membranes, but, at a minimum, only 19% of the hyperpolarization ofV bl could be accounted for by a furosemide-induced reduction in basolateral membrane Cl− current. Thus an increase in intracellular Cl− activity may have contributed to the depolarization ofV bl during net Cl− absorption, and the intracellular Cl− activity was likely greater with ADH than without hormone. Since ADH increases apical K+ conductance and since the chemical driving force for electroneutral Na+,K+,2Cl− cotransport from lumen to cell may have been less in the presence of ADH than in the absence of hormone, the cardinal effects of ADH may have been to increase the functional number of both Ba++-sensitive conductance K+ channels and electroneutral Na+,K+,2Cl− cotransport units in apical plasma membranes.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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  • 2
    ISSN: 1432-1424
    Schlagwort(e): renal tubule transport ; medullary thick ascending limb ; ADH ; K+ conductance ; Na+,K+,Cl− cotransport ; cell conductance
    Quelle: Springer Online Journal Archives 1860-2000
    Thema: Biologie , Chemie und Pharmazie
    Notizen: Summary This paper reports experiments designed to assess the relations between net salt absorption and transcellular routes for ion conductance in single mouse medullary thick ascending limbs of Henle microperfusedin vitro. The experimental data indicate that ADH significantly increased the transepithelial electrical conductance, and that this conductance increase could be rationalized in terms of transcellular conductance changes. A minimal estimate (G c min ) of the transcellular conductance, estimated from Ba++ blockade of apical membrane K+ channels, indicated thatG c min was approximately 30–40% of the measured transepithelial conductance. In apical membranes, K+ was the major conductive species; and ADH increased the magnitude of a Ba++-sensitive K+ conductance under conditions where net Cl− absorption was nearly abolished. In basolateral membranes, ADH increased the magnitude of a Cl− conductance; this ADH-dependent increase in basal Cl− conductance depended on a simultaneous hormone-dependent increase in the rate of net Cl− absorption. Cl− removal from luminal solutions had no detectable effect onG e , and net Cl− absorption was reduced at luminal K+ concentrations less than 5mm; thus apical Cl− entry may have been a Na+,K+,2Cl− cotransport process having a negligible conductance. The net rate of K+ secretion was approximately 10% of the net rate of Cl− absorption, while the chemical rate of net Cl− absorption was virtually equal to the equivalent short-circuit current. Thus net Cl− absorption was rheogenic; and approximately half of net Na+ absorption could be rationalized in terms of dissipative flux through the paracellular pathway. These findings, coupled with the observation that K+ was the principal conductive species in apical plasma membranes, support the view that the majority of K+ efflux from cell to lumen through the Ba++-sensitive apical K+ conductance pathway was recycled into cells by Na+,K+,2Cl− cotransport.
    Materialart: Digitale Medien
    Bibliothek Standort Signatur Band/Heft/Jahr Verfügbarkeit
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