Chloride Conductance and P
i
Transport are Separate Functions Induced by the Expression of NaPi-1 in Xenopus Oocytes |
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Authors: | S Bröer A Schuster CA Wagner A Bröer I Forster J Biber H Murer A Werner F Lang AE Busch |
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Institution: | Institute of Physiological Chemistry, Eberhard-Karls-Universit?t Tübingen, D-72076 Tübingen, Germany, DE Institute of Physiology, Eberhard-Karls-Universit?t Tübingen, D-72076 Tübingen, Germany, DE Institute of Physiology, University of Zürich, Winterhurerstrasse 190, CH-8057 Zürich, Switzerland, CH
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Abstract: | Expression of the protein NaPi-1 in Xenopus oocytes has previously been shown to induce an outwardly rectifying Cl− conductance (GCl), organic anion transport and Na+-dependent P
i
-uptake. In the present study we investigated the relation between the NaPi-1 induced GCl and P
i
-induced currents and transport. NaPi-1 expression induced P
i
-transport, which was not different at 1–20 ng/oocyte NaPi-1 cRNA injection and was already maximal at 1–2 days after cRNA
injection. In contrast, GCl was augmented at increased amounts of cRNA injection (1–20 ng/oocyte) and over a five day expression period. Subsequently
all experiments were performed on oocytes injected with 20 ng/oocytes cRNA. P
i
-induced currents (Ip) could be observed in NaPi-1 expressing oocytes at high concentrations of P
i
(≥ 1 mm P
i
). The amplitudes of Ip correlated well with GCl. Ip was blocked by the Cl− channel blocker NPPB, partially Na+-dependent and completely abolished in Cl− free solution. In contrast, P
i
-transport in NaPi-1 expressing oocytes was not NPPB sensitive, stronger depending on extracellular Na+ and weakly affected by Cl− substitution. Endogenous P
i
-uptake in water-injected oocytes amounted in all experiments to 30–50% of the Na+-dependent P
i
-transport observed in NaPi-1 expressing oocytes. The properties of the endogenous P
i
-uptake system (K
m
for P
i
> 1 mm; partial Na+- and Cl−-dependence; lack of NPPB block) were similar to the NaPi-1 induced P
i
-uptake, but no Ip could be recorded at P
i
-concentrations ≤3 mm. In summary, the present data suggest that Ip does not reflect charge transfer related to P
i
-uptake, but a P
i
-mediated modulation of GCl.
Received: 22 October 1997/Revised: 24 March 1998 |
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Keywords: | : Cl− Channel — Organic Anions — Phosphate — NPPB |
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