Synaptophysin is targeted to similar microvesicles in CHO and PC12 cells.
Synaptophysin, an integral membrane protein of small synaptic vesicles, was expressed by transfection in fibroblastic CHO-K1 cells. The properties and localization of synaptophysin were compared between transfected CHO-K1 cells and native neuroendocrine PC12 cells. Both cell types similarly glycosylate synaptophysin and sort it into indistinguishable microvesicles. These become labeled by endocytic markers and are primarily concentrated below the plasmalemma and at the area of the Golgi complex and the centrosomes. A small pool of synaptophysin is transiently found on the plasma membrane. In CHO-K1 cells synaptophysin co-localizes with transferrin that has been internalized by receptor-mediated endocytosis. These findings suggest that synaptophysin in transfected CHO-K1 cells and neuroendocrine PC12 cells is directed into a pathway of recycling microvesicles which, in CHO cells, is shown to coincide with that of the transferrin receptor. They further indicate that fibroblasts have the ability to sort a synaptic vesicle membrane protein. Our results suggest a pathway for the evolution of small synaptic vesicles from a constitutively recycling organelle which is normally present in all cells.
Johnston
P A
PA
Howard Hughes Medical Institute, UT Southwestern Medical Center, Dallas, TX 75235.
Cameron
P L
PL
Stukenbrok
H
H
Jahn
R
R
De Camilli
P
P
Südhof
T C
TC
D000310:Adrenal Gland Neoplasms / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D005455:Fluorescent Antibody Technique; D006031:Glycosylation; D015151:Immunoblotting; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D008854:Microscopy, Electron; D009419:Nerve Tissue Proteins / Q000032:analysis / Q000378:metabolism*; D009490:Neurosecretory Systems / Q000166:cytology / Q000378:metabolism*; D015388:Organelles / Q000032:analysis / Q000378:metabolism; D010673:Pheochromocytoma / Q000378:metabolism; D051381:Rats; D013572:Synaptic Vesicles / Q000378:metabolism*; D016708:Synaptophysin; D014162:Transfection; D014168:Transferrin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D016708:Synaptophysin; D014168:Transferrin