TY - JOUR
T1 - Fluorescence visualization of receptor-bound low density lipoprotein in human fibroblasts
AU - Anderson, Richard G W
AU - Goldstein, Joseph I.
AU - Brown, Michael S.
N1 - Funding Information:
This research was supported by USPHS Grant PO1 HL-20948 from the National Institutes of Health. We thank Brenda Shaw, Gloria Y. Brunschede, and Michael Gaisbauer for their expert technical assistance.
PY - 1980
Y1 - 1980
N2 - Previous 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 became larger in size and fewer in number, suggesting rapid fusion of endocytic vesicles. After internalization of LDL, new unoccupied receptors rapidly appeared on the cell surface where they were again distributed in linear arrays. In fibroblasts from a patient with the LDL internalization defect (J.D.), LDL receptors fail to be incorporated in coated pits. By immuno-fluorescence, LDL that was bound to these mutant receptors produced fluorescent foci that tended to be smaller and more numerous than those of normal fibroblasts. Moreover, the fluorescent foci of LDL
AB - Previous 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 became larger in size and fewer in number, suggesting rapid fusion of endocytic vesicles. After internalization of LDL, new unoccupied receptors rapidly appeared on the cell surface where they were again distributed in linear arrays. In fibroblasts from a patient with the LDL internalization defect (J.D.), LDL receptors fail to be incorporated in coated pits. By immuno-fluorescence, LDL that was bound to these mutant receptors produced fluorescent foci that tended to be smaller and more numerous than those of normal fibroblasts. Moreover, the fluorescent foci of LDL
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U2 - 10.3109/10799898009039253
DO - 10.3109/10799898009039253
M3 - Article
C2 - 6271950
AN - SCOPUS:0019123045
SN - 1079-9893
VL - 1
SP - 17
EP - 39
JO - Journal of Receptors and Signal Transduction
JF - Journal of Receptors and Signal Transduction
IS - 1
ER -