The Journal of General Physiology
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Published online January 14, 2008
doi:10.1085/jgp.200709891
The Journal of General Physiology, Vol. 131, No. 2, 117-124
The Rockefeller University Press, 0022-1295 $30.00
© 2008 Zhang et al.
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ARTICLE

Temperature Dependence of Fusion Kinetics and Fusion Pores in Ca2+-triggered Exocytosis from PC12 Cells



Zhen Zhang and Meyer B. Jackson

Department of Physiology and Molecular and Cellular Pharmacology Ph.D. Program, University of Wisconsin School of Medicine and Public Health, Madison, WI 53706

Correspondence to Meyer Jackson: mjackson{at}physiology.wisc.edu

The temperature dependence of Ca2+-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°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 ~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 suggests that after the fusion pore has started to expand, structural transitions of membrane components are no longer kinetically limiting. This indicates that the content of a vesicle is expelled completely after fusion pore expansion.


Abbreviation used in this paper: RMS, root-mean-square.


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Z. Zhang and M. B. Jackson
Temperature Dependence of Fusion Kinetics and Fusion Pores in Ca2+-triggered Exocytosis from PC12 Cells
J. Cell Biol., January 28, 2008; 180(2): i8 - i8.
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