Cytoplasmic surface ultrastructures of cardiac gap junctions as revealed by quick-freeze, deep-etch replicas

Rapid-freeze, deep-etch, rotary-shadow replica studies were performed to examine the cytoplasmic surface membrane of the cardiac gap junctions of rats, mice, and guinea pigs. In quick-frozen fresh cardiac muscles, while the nonjunctional cytoplasmic surfaces were covered with filamentous materials, the cytoplasmic surface membrane continuous with freeze-fractured gap junction plaques were relatively free of such filaments and revealed particulate patterns. After brief rinsing in high K buffer, gap junction membranes showed granular substructures resembling a tiled surface made of round tiles of various sizes. After prolonged rinsing for more than 20 min, however, cytoplasmic surfaces of gap junctions became less particulate but rather smooth. The particulate substructures observed in the rapid-freeze deep-etch replicas may correspond to the fuzzy cytoplasmic layer in thin sections and serine protease sensitive peptide moiety in sodium dodecyl sulfate-polyacrylamide gel electrophoresis reported in isolated cardiac gap junction pellets. These cytoplasmic components, which are absent in liver gap junctions, seem to be specific in cardiac and neural gap junctions and may be related to the large electrical current passed by these junctions.