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Biophys. J. BioFAST: First Published June 24, 2005. doi:10.1529/biophysj.105.061663
© 2005 by the Biophysical Society.

A more recent version of this article appeared on September 1, 2005.
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SPECTROSCOPY, IMAGING, OTHER TECHNIQUES

Visualization of synaptic vesicle movement in intact synaptic boutons using fluorescence fluctuation spectroscopy

Randolf Jordan 1, Edward A Lemke 1 and Jurgen Klingauf 1*

1 Max-Planck Institute for Biophysical Chemistry

* To whom correspondence should be addressed. E-mail: jklinga{at}gwdg.de.

Submitted on February 23, 2005
Revised on April 15, 2005
Accepted on 7 June 2005


  Abstract
Not much is known about the mobility of synaptic vesicles inside small synapses of the central nervous system, reflecting a lack of methods for visualizing these dynamics. We adapted confocal spot detection with fluctuation analysis to monitor the mobility of fluorescently labeled synaptic vesicles inside individual boutons of cultured hippocampal neurons. Using Monte-Carlo simulations we were able to propose a simple quantitative model that can quantitatively describe vesicle mobility in small hippocampal boutons under resting conditions and different pharmacological treatments. We find that vesicle mobility in a time window of 20 s is best described by caged diffusion (D ~ 5 10-5 ìm2/s, cage radii of ~50 nm). Mobility can be up-regulated by phosphatase blockage and increased further by actin disruption in a dose-dependent manner. Inhibition of the myosin light chain kinase (MLCK) slowed down vesicle mobility 10-fold, while other kinases like PKC, PKA and CaMKII do not affect mobility in unstimulated boutons.

Key Words: exo-endocytosis, fluorescence, presynaptic mechanisms, vesicle mobility




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Copyright © 2005 by the Biophysical Society.