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Am J Physiol Cell Physiol (December 18, 2003). doi:10.1152/ajpcell.00074.2003
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Submitted on February 25, 2003
Accepted on December 9, 2003

A voltage-gated K+ current in renal inner medullary collecting duct cells

Laura I Escobar1*, Julio C Martinez-Tellez1, Monica Salas1, Salvador A Castilla1, Roland Carrisoza1, Dagoberto Tapia2, Mario Vazquez1, Jose Bargas2, and Juan J Bolivar1

1 Departamento de Fisiologia, Facultad de Medicina, Universidad Nacional Autonoma de Mexico, Mexico, DF, Mexico
2 Departamento de Biofisica, Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, Mexico, DF, Mexico

* To whom correspondence should be addressed. E-mail: laurae{at}servidor.unam.mx.

We studied the K+ selective conductances in primary cultures of rat renal inner medullary collecting duct (IMCD) using perforated-patch and conventional whole cell techniques. Depolarizations above -20 mV induced a time-dependent outward K+ current (Ivto) similar to a delayed-rectifier. Ivto showed a half maximal activation around 5.6 mV with a slope factor of 6.8 mV. Its K+/Na+ selectivity ratio was 11.7. It was inhibited by TEA, quinidine, 4-aminopyridine and Ba2+, and was not Ca2+-dependent. The delayed rectifying characteristics of Ivto prompted us to screen the expression of Kv1 and Kv3 families by RT-PCR. Analysis of RNA isolated from cell cultures revealed the presence of three Kv {alpha}-subunits (Kv1.1, Kv1.3 and Kv1.6). Western blot analysis with Kv a-subunits antibodies for Kv1.1 and Kv1.3 showed labeling of about 70 kDa proteins from inner medulla plasmatic and microsome membranes. Immunocytochemical analysis of cell culture and kidney inner medulla showed that Kv1.3 is colocalized with the Na+/K+-ATPase at the basolateral membrane, although it is also in the cytoplasm. This is the first evidence of recording, protein expression and localization of a voltage-gated Kv1 in the kidney IMCD cells.




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