The permeability of isolated and in situ mouse hepatic gap junctions studied with enzymatic tracers.
We have studied the effects of phospholipase C from Clostridium welchii on gap junctions in the intact mouse liver and in a junction-rich fraction prepared from mouse liver. Treatment of the isolated junctions results in the disappearance of both the 20 A gap and of the polygonal lattice visible with lanthanum. The junctions are morphologically unaltered, however, when whole livers are perfused with phospholipase via the portal vein. These results suggest that extracellular phospholipase cannot diffuse into the junctional area, but that the enzyme may affect structures within the gap from its cytoplasmic surfaces which become exposed in the isolated preparations. Horseradish peroxidase, which has physical dimensions similar to those of Clostridium phospholipase is also denied access to the 20 A gap in whole liver, while peroxidase reaction product can be seen in the gap in isolated preparations. Beef liver catalase, however, a tracer molecule much larger than peroxidase, cannot penetrate even in isolated fractions. If the cytoplasmic approaches to the gap junction used by peroxidase and phospholipase are available in vivo, and have not been created during the process of mechanical isolation, they may play a role in cell-to-cell passage of molecules larger than ions.
D000818:Animals; D002374:Catalase / Q000494:pharmacology*; D002458:Cell Fractionation; D002463:Cell Membrane Permeability / Q000187:drug effects*; D003013:Clostridium; D005615:Freezing; D006651:Histocytochemistry; D007365:Intercellular Junctions / Q000187:drug effects; D007811:Lanthanum; D008099:Liver / Q000166:cytology*; D051379:Mice; D008854:Microscopy, Electron; D010447:Peptide Hydrolases / Q000494:pharmacology; D010544:Peroxidases / Q000494:pharmacology*; D010740:Phospholipases / Q000494:pharmacology*
D007811:Lanthanum; D010544:Peroxidases; D002374:Catalase; D010740:Phospholipases; D010447:Peptide Hydrolases