Fluorescence visualization of receptor-bound low density lipoprotein in human fibroblasts.

AbstractPrevious electron microscopic studies have shown that receptors for plasma low density lipoprotein (LDL) in human fibroblasts are located predominantly in coated pits on the cell surface. In the current studies, LDL receptors were visualized at the light microscopic level by two methods: 1) indirect immuno-fluorescent staining of receptor-bound LDL, and 2) direct visualization of LDL particles that were reconstituted with a fluorescent cholesteryl ester derivative. By both techniques, LDL receptors were seen to be clustered in minute foci that were aligned in linear arrays on the cell surface. These arrays corresponded to linear arrays of coated pits that were visualized by immunofluorescent staining of the coat protein, clathrin. During the internalization of receptor-bound LDL, each fluorescent focus of LDL particles seemed to enter the cell as an individual unit without the formation of large patches. Immediately after the reconstituted fluorescent LDL entered the cell, the fluorescent dots bec...

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