Release of endosomal content induced by plasma membrane tension: video image intensification time lapse analysis.
Rhodamine-labeled vinculin microinjected into chicken embryo fibroblasts and rhodamine-labeled alpha 2-macroglobulin added to the fibroblast culture medium were sequestered in endocytotic vesicles and digested. When a sealed microcapillary coated with fibronectin or polylysine was attached to the fibroblasts and pulled at speeds of 100-200 microns/h, stretching the plasma membrane, a variable fraction of the endosomes released the rhodamine label. Release from individual vesicles was rapid, reaching completion in less than 30 to 120 s. The microfilament disrupting agent cytochalasin B prevented release, as did the microtubule stabilizing drug taxol. Colcemide, which disrupts microtubules, did not inhibit the release. Release was dependent on extracellular calcium, as it was prevented by 10 mM EGTA in the incubation medium. We postulate that opening of vesicular channels consequent to centripetal transmission of tension generated in the plasma membrane along microfilaments may be a mechanism of release of endosomal content.
Friedrich Miescher Institut, Basel, Switzerland.
J
J
Hagmann
D
D
Dagan
M M
MM
Burger
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D002462:Cell Membrane / Q000502:physiology*; D002642:Chick Embryo; D003571:Cytochalasin B / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D007089:Image Enhancement; D008247:Lysosomes / Q000378:metabolism*; D008845:Microinjections; D008856:Microscopy, Fluorescence; D017239:Paclitaxel; D013314:Stress, Mechanical; D014741:Video Recording
D000470:Alkaloids; D003571:Cytochalasin B; D004533:Egtazic Acid; D017239:Paclitaxel; D002118:Calcium; D003703:Demecolcine