Lysosomal uptake of isolated cell organelles microinjected into HeLa cells.
Lysosomes and microsomes were isolated from rat liver and microinjected into the cytoplasm of HeLa cells. The fate of the transplanted organelles and their effects on the recipient cells were followed in the electron microscope at various time intervals after administration. Needle injection with buffer or sucrose did not seem to evoke any ultrastructural alterations, such as induced autophagy or other signs of sublethal cell injury. Recipients of microinjected cell organelles elicited a rapid and conspicuous increase in membrane-bounded cytoplasmic vacuoles, concomitant with the disappearance of the injected material. Golgi complexes became abundant with many small vesicles clustering around their cisternae. The volume density of the lysosomal compartment increased 2-3-fold after organelle injection as compared with control-injected (0.3 M sucrose) or noninjected cells. Our preliminary results show that isolated cell organelles can be microinjected into cells n culture and indicate that the microinjected organelles were segregated from the cytoplasm into membrane-bounded vacuoles probably through autophagolysosome formation. Thus, this technique offers an additional approach for studies on the segregation and degradation of cell organelles in somatic cells and may enable more detailed analyses on the mechanisms of autophagic sequestration of specific cell organelles.
D000818:Animals; D002462:Cell Membrane / Q000648:ultrastructure; D003593:Cytoplasm / Q000648:ultrastructure; D006367:HeLa Cells / Q000648:ultrastructure; D006801:Humans; D008099:Liver / Q000648:ultrastructure; D008247:Lysosomes / Q000648:ultrastructure*; D008845:Microinjections; D008854:Microscopy, Electron; D008862:Microsomes, Liver / Q000648:ultrastructure*; D009940:Organoids / Q000648:ultrastructure*; D051381:Rats; D014617:Vacuoles / Q000648:ultrastructure*