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The Journal of Neuroscience, October 15, 2001, 21(20):7928-7936
Protein Kinase C Increases the Apparent Affinity of the Release Machinery to Ca2+ by Enhancing the Release Machinery Downstream of the Ca2+ Sensor
Xin-Sheng Wu and Ling-Gang Wu Departments of Anesthesiology and Anatomy and Neurobiology, Washington University School of Medicine, St. Louis, Missouri 63110
Modulation of the release probability of releasable vesicles in response to Ca2+ influx (ProbCa) is involved in mediating several forms of synaptic plasticity, including short-term depression, short-term augmentation, and potentiation induced by protein kinases. Given such an important role, however, the mechanism underlying modulation of the ProbCa is unclear. We addressed this question by investigating how the activation of protein kinase C modulates the ProbCa at a calyx-type nerve terminal in rat brainstem. Various lengths of step depolarization were applied to the nerve terminal to evoke different amounts of Ca2+ currents and capacitance jumps, the latter of which reflect vesicle release. The relationship between the capacitance jump and the Ca2+ current integral was sigmoidal and was fit well with a Hill function. The sigmoidal relationship was shifted significantly to the left during the application of the PKC activator 12-myristate 13-acetate (PMA), suggesting that PMA increases the apparent affinity of the release machinery to Ca2+. This effect was blocked in large part by the application of the PKC inhibitor bisindolylmaleimide, suggesting that the effect is mediated mainly by the activation of PKC. We also found that PMA increased the rate of miniature EPSCs evoked by the application of hypertonic sucrose solution, which triggers release downstream of the Ca2+ influx. Taken together, our results suggest that PKC enhances the apparent affinity of the release machinery to Ca2+ by a mechanism downstream of the binding between Ca2+ and its sensor. These results have provided the first example of the mechanisms underlying modulation of the ProbCa.
Key words: protein kinase C; calyx of Held; release probability; capacitance measurement; releasable pool; vesicle mobilization
Copyright © 2001 Society for Neuroscience 0270-6474/01/21207928-09$05.00/0
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