Temperature dependence of fusion kinetics and fusion pores in Ca2+-triggered exocytosis from PC12 cells

The Journal of General Physiology
Zhen Zhang, Meyer B Jackson

Abstract

The temperature dependence of Ca(2+)-triggered exocytosis was studied using carbon fiber amperometry to record the release of norepinephrine from PC12 cells. Single-vesicle fusion events were examined at temperatures varying from 12 to 28 degrees C, and with release elicited by depolarization. Measurements were made of the initial and maximum frequencies of exocytotic events, of fusion pore lifetime, flux through the open fusion pore, kiss-and-run versus full-fusion probability, and parameters associated with the shapes of amperometric spikes. The fusion pore open-state flux, and all parameters associated with spike shape, including area, rise time, and decay time, had weak temperature dependences and activation energies in the range expected for bulk diffusion in an aqueous solution. Kiss-and-run events also varied with temperature, with lower temperatures increasing the relative probability of kiss-and-run events by approximately 50%. By contrast, kinetic parameters relating to the frequency of exocytotic events and fusion pore transitions depended much more strongly on temperature, suggesting that these processes entail structural rearrangements of proteins or lipids or both. The weak temperature dependence of spike shape su...Continue Reading

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Citations

Sep 17, 2008·Pflügers Archiv : European journal of physiology·Salma T HannaPatrick E MacDonald
Jul 4, 2008·Nature Structural & Molecular Biology·Meyer B Jackson, Edwin R Chapman
Feb 6, 2010·Proceedings of the National Academy of Sciences of the United States of America·Erdem KaratekinJames E Rothman
Oct 16, 2009·Molecular Biology of the Cell·Zhen ZhangMeyer B Jackson
Jun 25, 2010·Molecular Biology of the Cell·Zhen ZhangMeyer B Jackson
May 3, 2013·The Journal of Neuroscience : the Official Journal of the Society for Neuroscience·Ana Isabel CalejoRobert Zorec
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Feb 9, 2017·The Journal of General Physiology·Che-Wei ChangMeyer B Jackson
Feb 1, 2019·Proceedings of the National Academy of Sciences of the United States of America·Fabio MancaMatthieu Caruel
Jun 2, 2010·Biophysical Journal·Zhen Zhang, Meyer B Jackson
Sep 28, 2010·Journal of Neuroscience Methods·Reut Friedrich, Uri Ashery

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