Diffusion of injected macromolecules within the cytoplasm of living cells.
The diffusion of macromolecules introduced into the cytoplasm of human fibroblasts by erythrocyte-mediated microinjection was measured by the fluorescence recovery after photobleaching technique. The apparent diffusion coefficients for fluorescein-labeled IgG and fluorescein-labeled bovine serum albumin were approximately 10(-8) cm2/sec at 22 degrees C, consistent with the kinetics of spreading of the fluorescent probe following microinjection and approximately 1/70 the values in aqueous buffer. The diffusion of labeled bovine serum albumin was shown to be strongly dependent on temperature and, in fact, similar to that expected in a 61% aqueous sucrose solution. However, the marked reduction in diffusion at 5 degrees C could be fully reversed by incubation with 0.1 mM colchicine. These findings suggest that cytoplasmic diffusion rates are reduced relative to rates in aqueous media as a result of increased aqueous phase viscosity or the impedence provided by structural elements. Several simple models to account for the data are presented.
D002478:Cells, Cultured; D003078:Colchicine / Q000494:pharmacology; D003593:Cytoplasm / Q000502:physiology / Q000648:ultrastructure*; D004058:Diffusion; D005452:Fluoresceins; D006801:Humans; D007074:Immunoglobulin G; D008297:Male; D010777:Photochemistry; D012710:Serum Albumin, Bovine; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014783:Viscosity
D005452:Fluoresceins; D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D003078:Colchicine