Davis L M, Kanter H L, Beyer E C, Saffitz J E
Department of Pediatrics, Washington University School of Medicine, Saint Louis, Missouri 63110.
J Am Coll Cardiol. 1994 Oct;24(4):1124-32. doi: 10.1016/0735-1097(94)90879-6.
We sought to characterize the connexin phenotypes of selected regions of the canine heart with different conduction properties to determine whether variations in connexin expression might contribute to the differences in intercellular resistance and conduction velocity that occur in different cardiac tissues.
Gap junctions connect cardiac myocytes, allowing propagation of action potentials. Intercellular channels with different electrophysiologic properties are formed by different connexin proteins.
To determine which connexins were likely to be expressed in the sinus node, atrioventricular (AV) node and atrial and ventricular myocardium, messenger ribonucleic acids (RNAs) from each of these sites were hybridized with probes for connexin26, connexin31, connexin32, connexin37, connexin40, connexin43, connexin45, connexin46 and connexin50. Immunostaining with monospecific antibodies to connexin40, connexin43 and connexin45 was used to delineate the distribution of connexins in frozen sections of these different cardiac tissues.
Only messenger RNAs coding for connexin40, connexin43 and connexin45 were detected by Northern blot analysis. By immunohistochemical staining, junctions in the sinus and AV nodes and proximal His bundle were virtually devoid of connexin43 but contained both connexin40 and connexin45. Gap junctions in the distal His bundle and the proximal bundle branches stained intensely for connexin40 and connexin43 and to a lesser extent for connexin45. Atrial gap junctions showed abundant staining of connexin43, connexin40 and connexin45. Ventricular gap junctions were characterized by abundant staining of connexin43 and connexin45 and much less staining of connexin40.
Although most cardiac gap junctions contain connexin40, connexin43 and connexin45, the relative amounts of each of these connexins vary considerably in cardiac tissues with different conduction properties.
我们试图对犬心脏中具有不同传导特性的选定区域的连接蛋白表型进行表征,以确定连接蛋白表达的变化是否可能导致不同心脏组织中细胞间电阻和传导速度的差异。
缝隙连接连接心肌细胞,允许动作电位的传播。具有不同电生理特性的细胞间通道由不同的连接蛋白形成。
为了确定哪些连接蛋白可能在窦房结、房室(AV)结以及心房和心室心肌中表达,将来自这些部位的信使核糖核酸(RNA)与连接蛋白26、连接蛋白31、连接蛋白32、连接蛋白37、连接蛋白40、连接蛋白43、连接蛋白45、连接蛋白46和连接蛋白50的探针进行杂交。使用针对连接蛋白40、连接蛋白43和连接蛋白45的单特异性抗体进行免疫染色,以描绘这些不同心脏组织冰冻切片中连接蛋白的分布。
通过Northern印迹分析仅检测到编码连接蛋白40、连接蛋白43和连接蛋白45的信使RNA。通过免疫组织化学染色,窦房结和房室结以及希氏束近端的连接处几乎没有连接蛋白43,但同时含有连接蛋白40和连接蛋白45。希氏束远端和束支近端的缝隙连接对连接蛋白40和连接蛋白43染色强烈,但对连接蛋白45染色较弱。心房缝隙连接显示连接蛋白43、连接蛋白40和连接蛋白45染色丰富。心室缝隙连接的特征是连接蛋白43和连接蛋白45染色丰富,但连接蛋白40染色较少。
虽然大多数心脏缝隙连接含有连接蛋白40、连接蛋白43和连接蛋白45,但这些连接蛋白在具有不同传导特性的心脏组织中的相对含量差异很大。