Early membrane retrieval following exocytosis in rat mast cells.

The fate of the surplus membrane following exocytosis of mast cell granules was studied by the extracellular tracer Ruthenium red (Ru red). Isolated rat peritoneal mast cells were stimulated with 4 micrograms/ml polylysine, washed and maintained in a culture medium for 80 min. Mast cells were observed both with the light microscope after adding Ru red in physiological solution and with the electron microscope after fixation in Ru red-containing fixatives. Whereas all exocytotic cavities were found to be stained with Ru red immediately after stimulation, a gradual lack of staining was observed in the subsequent period. The exits of the cavities were sealed by membrane fusions which resulted in closed vacuoles containing exocytosed granule remnants. These vacuoles often fused with each other to form a few giant vacuoles. The overwhelming majority of the vacuoles were observed to be closed 30 to 80 min after stimulation. In one experiment a quantitative analysis was performed to assess the degree of membrane recapture by sealing of the exocytotic cavities. A considerable portion of the plasma membrane area was retrieved in this way as early as between 15 and 30 min after stimulation. We conclude that the dominant mechanism of membrane retrieval in the early period following exocytosis is the recapture of large membrane areas by sealing of exocytotic cavities.