Localization of gap junction proteins, connexins 32 and 26, in rat and guinea pig liver as revealed by quick-freeze, deep-etch immunoelectron microscopy

By use of site-specific antibodies against synthetic oligopeptides, we examined the localizations of the gap junction proteins connexin 32 (Cx32) and connexin 26 (Cx26) in rat and guinea pig liver. Double-labeling immunofluorescence microscopy revealed that in guinea pig liver both proteins were spread throughout the liver lobules and seemed to localize together within the same gap junction plaque. In rat liver, co-localization of both Cx32 and Cx26 in the same plaques was also suggested in periportal zones. Quick-freeze, deep-etch immunoelectron microscopy showed that immunolabeling of isolated guinea pig liver gap junction plaques with either Cx32 or Cx26 antiserum yielded complete and dense antibody decoration of the cytoplasmic surface of the plaques. In isolated rat liver plaques, the cytoplasmic surfaces were densely decorated with Cx32 antiserum, whereas Cx26 labeling yielded diffuse decoration with variable intensity of the plaques. In both species we did not observe any focal or patchy clusters of the labeling in any plaques examined. Double-labeling immunoelectron microscopy confirmed that both Cx32 and Cx26 are co-localized in the same gap junction plaques. These results suggest that in hepatocytes expressing both Cx32 and Cx26, both types of gap junction proteins are not segregated but intermingle randomly within the same plaques.