Cloning, expression, and localization of a rat hepatocyte inwardly rectifying potassium channel.
Bile formation involves anion accumulation within the apical lumen of hepatocytes. Potassium flux through hepatocellular basolateral membrane channels may provide the counterion for apical anion efflux. Here we cloned a molecular candidate for maintaining charge balance during bile secretion. Two transcripts resembling the Kir4.2 subclass of inwardly rectifying potassium channels were found. The longer deduced isoform (4.2a) has 30 additional NH(3)-terminal amino acids, which identifies this as a new isoform. The short-form isoform shared 86-91% identity with the mouse, human, and guinea pig channels. Whole cell currents of either rat isoform expressed in HEK293T cells demonstrated time independence and inward rectification. Antibodies against a COOH-terminal fragment recognized bands between 40 and 45 kDa and at 90 kDa and recognized a high molecular mass band around 200 kDa in overexpressing HEK cells. Immunohistology of liver tissue shows hepatocellular plasma membrane localization. In hepatocyte couplets, Kir4.2 was predominantly localized to the basolateral membrane. Results demonstrate expression of a new Kir4.2 isoform in the rat hepatocyte whose functional properties are compatible with a role in maintaining electrical integrity of bile-generating hepatocytes.
Hill
Ceredwyn E
CE
Gastrointestinal Diseases Research Unit and Department of Physiology, Queen's University, Kingston, Ontario K7L 5G2, Canada. hillc@post.queensu.ca
Briggs
M Martha
MM
Liu
Junjun
J
Magtanong
Leslie
L
D000818:Animals; D000906:Antibodies; D000918:Antibody Specificity; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D015870:Gene Expression / Q000502:physiology; D022781:Hepatocytes / Q000737:chemistry* / Q000502:physiology*; D006801:Humans; D007668:Kidney / Q000166:cytology; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008969:Molecular Sequence Data; D018408:Patch-Clamp Techniques; D024661:Potassium Channels, Inwardly Rectifying / Q000032:analysis / Q000235:genetics* / Q000276:immunology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D015398:Signal Transduction / Q000502:physiology; D014162:Transfection
D000906:Antibodies; D024661:Potassium Channels, Inwardly Rectifying; D012333:RNA, Messenger