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A more recent version of this article appeared on January 1, 2003
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Submitted on January 29, 2002
Revised on September 12, 2002
Accepted on September 30, 2002
1 Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3200 (present address: Misaki Marine Biological Station, University of Tokyo, Misaki, Miura, Kanagawa 238-0225, Japan)
2 Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3200
* Corresponding author. E-mail address: rsteinha{at}socrates.berkeley.edu.
We previously found that a micro-disruption of the plasma membrane evokes Ca2+-regulated exocytosis near the wound site, which is essential for membrane resealing. We demonstrate here that repeated membrane disruption reveals long-term potentiation of Ca2+-regulated exocytosis in 3T3 fibroblasts, which is closely correlated with faster membrane resealing rates. This potentiation of exocytosis is cAMP-dependent protein kinase (PKA)-dependent in the early stages (minutes), in the intermediate-term (hrs.) requires protein synthesis, and for long-term (24 hrs) depends on the activation of cAMP responsive element-binding protein (CREB). We were able to demonstrate that wounding cells activated CREB within 3.5 hrs. In all three phases the increase in the amount of exocytosis was correlated with an increase in the rate of membrane resealing. However, a brief treatment with forskolin, which is effective for short-term potentiation and which could also activate CREB, was not sufficient to induce long-term potentiation of resealing. These results imply that long-term potentiation by CREB required activation by another, cAMP-independent, pathway.
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