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Am J Physiol Cell Physiol 294: C893-C906, 2008. First published January 16, 2008; doi:10.1152/ajpcell.00616.2006
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MEMBRANE TRANSPORTERS, ION CHANNELS, AND PUMPS

A hyperpolarization-activated, cyclic nucleotide-gated, (Ih-like) cationic current and HCN gene expression in renal inner medullary collecting duct cells

Juan J. Bolívar,1 Dagoberto Tapia,2 Gabina Arenas,3 Mauricio Castañón-Arreola,4 Haydee Torres,3 and Elvira Galarraga2

1Departamento de Fisiología, Facultad de Medicina, 2Departamento de Biofísica, Instituto de Fisiología Celular, 3Departamento de Microbiología, Facultad de Medicina, Universidad Nacional Autónoma de México, Cuidad de México; and 4Posgrado en Ciencias Genómicas, Universidad Autónoma de la Ciudad de México, Ciudad de México, Mexico

Submitted 11 December 2006 ; accepted in final form 11 January 2008

The cation conductancein primary cultures of rat renal inner medullary collecting duct was studied using perforated-patch and conventional whole cell clamp techniques. Hyperpolarizations beyond –60 mV induced a time-dependent inward nonselective cationic current (Ivti) that resembles the well-known hyperpolarization-activated, cyclic nucleotide-gated Ih and If currents. Ivti showed a half-maximal activation around –102 mV with a slope factor of 25 mV. It had a higher conductance (but, at its reversal potential, not a higher permeability) for K+ than for Na+ (gK+/gNa+ = 1.5), was modulated by cAMP and blocked by external Cd2+ (but not Cs+ or ZD-7288), and potentiated by a high extracellular K+ concentration. We explored the expression of the Ih channel genes (HCN1 to -4) by RT-PCR. The presence of transcripts corresponding to the HCN1, -2, and -4 genes was observed in both the cultured cells and kidney inner medulla. Western blot analysis with HCN2 antibody showed labeling of ~90- and ~120-kDa proteins in samples from inner medulla and cultured cells. Immunocytochemical analysis of cell cultures and inner medulla showed the presence of HCN immunoreactivity partially colocalized with the Na+-K+-ATPase at the basolateral membrane of collecting duct cells. This is the first evidence of an Ih-like cationic current and HCN immunoreactivity in either kidney or any other nonexcitable mammalian cells.

kidney; hyperpolarization-activated current; nonselective cation channel; sodium transport



Address for reprint requests and other correspondence: J. J. Bolívar, Departamento de Fisiología, Facultad de Medicina, Universidad Nacional Autónoma de México, P. O. Box 70-250, México City DF, 04510, México (e-mail: jjboliv{at}servidor.unam.mx)







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