This Article Full Text Full Text (PDF) supplemental All Versions of this Article: biophysj.105.061663v1 89/3/2091    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Jordan, R. Articles by Klingauf, J. Search for Related Content PubMed PubMed Citation Articles by Jordan, R. Articles by Klingauf, J.

Visualization of Synaptic Vesicle Movement in Intact Synaptic Boutons Using Fluorescence Fluctuation Spectroscopy

Max-Planck Institute for Biophysical Chemistry, Department of Membrane Biophysics, Am Fassberg 11, 37077 Goettingen, Germany

Correspondence: Address reprint requests to Dr. Jurgen Klingauf, Dept. of Membrane Biophysics, Max-Planck Institute for Biophysical Chemistry, Am Fassberg 11, 37077 Goettingen, Germany. Tel.: 49-551-201-1629; Fax: 49-551-201-1688; E-mail: J.Klingauf{at}tmpi-bpc.mpg.de.

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 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 can be well described by caged diffusion (D 5x10^–5 µm²/s, cage sizes of 50 nm). Mobility can be upregulated by phosphatase blockage and increased further by actin disruption in a dose-dependent manner. Inhibition of the myosin light chain kinase slows down vesicle mobility 10-fold, whereas other kinases like protein kinase C (PKC), A (PKA), and calmodulin kinase II (caMKII) do not affect mobility in unstimulated boutons.

This article has been cited by other articles:

				V. Westphal, S. O. Rizzoli, M. A. Lauterbach, D. Kamin, R. Jahn, and S. W. Hell Video-Rate Far-Field Optical Nanoscopy Dissects Synaptic Vesicle Movement Science, April 11, 2008; 320(5873): 246 - 249. [Abstract] [Full Text] [PDF]

				G. Srinivasan, J. H. Kim, and H. von Gersdorff The Pool of Fast Releasing Vesicles Is Augmented by Myosin Light Chain Kinase Inhibition at the Calyx of Held Synapse J Neurophysiol, April 1, 2008; 99(4): 1810 - 1824. [Abstract] [Full Text] [PDF]

				V. Jensen, S. I. Walaas, S. Hilfiker, A. Ruiz, and O. Hvalby A delayed response enhancement during hippocampal presynaptic plasticity in mice J. Physiol., August 15, 2007; 583(1): 129 - 143. [Abstract] [Full Text] [PDF]

				C. Yeung, M. Shtrahman, and X.-l. Wu Stick-and-Diffuse and Caged Diffusion: A Comparison of Two Models of Synaptic Vesicle Dynamics Biophys. J., April 1, 2007; 92(7): 2271 - 2280. [Abstract] [Full Text] [PDF]

				H. Tokuoka and Y. Goda Myosin Light Chain Kinase Is Not a Regulator of Synaptic Vesicle Trafficking during Repetitive Exocytosis in Cultured Hippocampal Neurons. J. Neurosci., November 8, 2006; 26(45): 11606 - 11614. [Abstract] [Full Text] [PDF]

				E. A. Lemke and J. Klingauf Single Synaptic Vesicle Tracking in Individual Hippocampal Boutons at Rest and during Synaptic Activity J. Neurosci., November 23, 2005; 25(47): 11034 - 11044. [Abstract] [Full Text] [PDF]