Plasma membrane domains specialized for clathrin-mediated endocytosis in primary cells
Department of Physiology; Program in Molecular Medicine
3T3-L1 Cells; Animals; Cells, Cultured; Clathrin; Endocytosis; Fluorescent Dyes; Male; Mice; Mice, Inbred C57BL
Life Sciences | Medicine and Health Sciences
Clathrin assembly at the plasma membrane is a fundamental process required for endocytosis. In cultured cells, most of the clathrin is localized to large patches that display little lateral mobility. The functional role of these regions is not clear, and it has been thought that they may represent artifacts of cell adhesion of cultured cells. Here we have analyzed clathrin organization in primary adipose cells isolated from mice, which are nonadherent and fully differentiated. The majority of clathrin on the plasma membrane of these cells (>60%) was found in large clathrin patches that displayed virtually no lateral mobility and persisted for many minutes, and a smaller amount was found in small spots that appeared and disappeared rapidly. Direct visualization of transferrin revealed that it bound onto large arrays of clathrin, internalizing through vesicles that emerge from these domains. High resolution imaging (50 images/s) revealed fluorescence intensity fluctuations consistent with the formation and detachment of coated vesicles from within large patches. These results reveal that large clathrin assemblies are active regions of endocytosis in mammalian cells and highlight the importance of understanding the mechanistic basis for this organization.
DOI of Published Version
J Biol Chem. 2006 Jun 9;281(23):16139-46. Epub 2006 Mar 13. Link to article on publisher's site
The Journal of biological chemistry
Bellve, Karl D.; Leonard, Deborah; Standley, Clive; Lifshitz, Lawrence M.; Tuft, Richard A.; Hayakawa, Akira; Corvera, Silvia; and Fogarty, Kevin E., "Plasma membrane domains specialized for clathrin-mediated endocytosis in primary cells" (2006). Open Access Articles. 681.