Differential distribution of Kir2.1 and Kir2.3 subunits in canine atrium and ventricle.
Ventricular inward rectifier K(+) current (I(K1)) is substantially larger than atrial, producing functionally important action potential differences. To evaluate possible molecular mechanisms, we recorded I(K1) with patch-clamp techniques and studied Kir2.1 and Kir2.3 subunit expression. I(K1) density was >10-fold larger in the canine ventricle than atrium. Kir2.1 protein expression (Western blot) was 78% greater (P < 0.01) in the ventricle, but Kir2.3 band density was 228% greater (P < 0.01) in the atrium. Immunocytochemistry showed transverse tubular localization of Kir2.1 in 89% (17 of 19) of ventricular and 26% (5 of 19, P < 0.0001) of atrial cells. Both exhibited a weakly positive Kir2.1 signal at intercalated disks. Kir2.3 was strongly expressed at the intercalated disks in all cells and in the transverse tubular regions in 78% (14 of 18) of atrial and 22% (4 of 18, P < 0.001) of ventricular cells. Tissue immunohistochemical results qualitatively resembled isolated cell data. We conclude that the expression density and subcellular localization of Kir2.1 and Kir2.3 subunits differ in the canine atrium versus ventricle. Overall protein density differences are insufficient to explain I(K1) discrepancies, which may be related to differences in subcellular distribution.
Department of Medicine and Research Center, Montreal Heart Institute, Montreal, Quebec H1T 1C8, Canada.
Peter
P
Melnyk
Liming
L
Zhang
Alvin
A
Shrier
Stanley
S
Nattel
D000818:Animals; D015153:Blotting, Western; D004285:Dogs; D006321:Heart / Q000502:physiology; D006325:Heart Atria / Q000737:chemistry / Q000166:cytology; D006352:Heart Ventricles / Q000737:chemistry / Q000166:cytology; D007150:Immunohistochemistry; D008564:Membrane Potentials / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000737:chemistry / Q000502:physiology; D009206:Myocardium / Q000737:chemistry* / Q000166:cytology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000032:analysis / Q000502:physiology; D024661:Potassium Channels, Inwardly Rectifying / Q000032:analysis* / Q000502:physiology*
D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying