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MBC in Press, published online ahead of print July 11, 2003
Mol. Biol. Cell 10.1091/mbc.E03-01-0050

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Submitted on January 28, 2003
Revised on May 29, 2003
Accepted on June 9, 2003

Cell surface orifices of caveolae and localization of caveolin to the necks of caveolae in adipocytes

Hans Thorn1, Karin G. Stenkula1, Margareta Karlsson1, Unn Örtegren1, Fredrik H. Nystrom1, Johanna Gustavsson1, and Peter Strålfors1*

1 Department of Cell Biology, Faculty of Health Sciences, Linköping University, SE58185 Linköping, Sweden

* Corresponding author. E-mail address: peter.stralfors{at}ibk.liu.se.

Caveolae are noncoated invaginations of the plasma membrane that form in the presence of the protein caveolin. Caveolae are found in most cells, but are especially abundant in adipocytes. By high-resolution electron microscopy of plasma membrane sheets the detailed structure of individual caveolae of primary rat adipocytes was examined. Caveolin-1 and -2 binding was restricted to the membrane proximal region, such as the ducts or necks attaching the caveolar bulb to the membrane. This was confirmed by transfection with myc-tagged caveolin-1 and -2. Essentially the same results were obtained with human fibroblasts. Hence caveolin does not form the caveolar bulb in these cells, but rather the neck and may thus act to retain the caveolar constituents, indicating how caveolin participates in the formation of caveolae. Caveolae, randomly distributed overthe plasma membrane, were very heterogeneous varying in size between 25 and 150 nm. There was about one million caveolae in an adipocyte, which increased the surface area of the plasma membrane by 50%. Half of the caveolae, those larger than 50 nm, had access to the out-side of the cell via ducts and 20 nm orifices at the cell surface. The rest of the caveolae, those smaller than 50 nm, were not open to the cell exterior. Cholesterol depletion destroyed both caveolae and the cell surface orifices.




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