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© The Rockefeller University Press, 0021-9525/2000/2/755/ $5.00
The Journal of Cell Biology, Volume 148, Number 4, February 21, 2000 755-768

Original Article

Exocytotic Insertion of Calcium Channels Constrains Compensatory Endocytosis to Sites of Exocytosis

Robert M. Smitha, Boris Baibakova, Yoshihide Ikebuchia, Benjamin H. Whiteb, Nevin A. Lamberta,c, Leonard K. Kaczmarekb, and Steven S. Vogela
a Medical College of Georgia, Augusta, Georgia 30912-2630
b Yale University School of Medicine, New Haven, Connecticut 06510
c Veterans Affairs Medical Center, Augusta, Georgia 30912-2630

Correspondence to: Steven S. Vogel, Medical College of Georgia, Institute of Molecular Medicine and Genetics, 1120 15th Street CB2803, Augusta, GA 30912-2630. Tel:(706) 721-1604 Fax:(706) 721-7915 E-mail:stevev{at}immag.mcg.edu.

Proteins inserted into the cell surface by exocytosis are thought to be retrieved by compensatory endocytosis, suggesting that retrieval requires granule proteins. In sea urchin eggs, calcium influx through P-type calcium channels is required for retrieval, and the large size of sea urchin secretory granules permits the direct observation of retrieval. Here we demonstrate that retrieval is limited to sites of prior exocytosis. We tested whether channel distribution can account for the localization of retrieval at exocytotic sites. We find that P-channels reside on secretory granules before fertilization, and are translocated to the egg surface by exocytosis. Our study provides strong evidence that the transitory insertion of P-type calcium channels in the surface membrane plays an obligatory role in the mechanism coupling exocytosis and compensatory endocytosis.

Key Words: agatoxin, conotoxin, P-type calcium channels, sea urchins, microscopy


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