Differential distribution of release-related proteins in the hippocampal CA3 area as revealed by freeze-fracture replica labeling

A. Hagiwara, Y. Fukazawa, M. Deguchi Tawarada, T. Ohtsuka, R. Shigemoto, Journal of Comparative Neurology 489 (2005) 195–216.

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Journal Article | Published
Author
Hagiwara, Akari; Fukazawa, Yugo; Deguchi-Tawarada, Maki; Ohtsuka, Toshihisa; Shigemoto, RyuichiIST Austria
Abstract
Synaptic vesicle release occurs at a specialized membrane domain known as the presynaptic active zone (AZ). Several membrane proteins are involved in the vesicle release processes such as docking, priming, and exocytotic fusion. Cytomatrix at the active zone (CAZ) proteins are structural components of the AZ and are highly concentrated in it. Localization of other release-related proteins including target soluble N-ethylmaleimide-sensitive-factor attachment protein receptor (t-SNARE) proteins, however, has not been well demonstrated in the AZ. Here, we used sodium dodecyl sulfate-digested freeze-fracture replica labeling (SDS-FRL) to analyze quantitatively the distribution of CAZ and t-SNARE proteins in the hippocampal CA3 area. The AZ in replicated membrane was identified by immunolabeling for CAZ proteins (CAZ-associated structural protein [CAST] and Bassoon). Clusters of immunogold particles for these proteins were found on the P-face of presynaptic terminals of the mossy fiber and associational/commissural (AJC) fiber. Co-labeling with CAST revealed distribution of the t-SNARE proteins syntaxin and synaptosomal-associated protein of 25 kDa (SNAP-25) in the AZ as well as in the extrasynaptic membrane surrounding the AZ (SZ). Quantitative analysis demonstrated that the density of immunoparticles for CAST in the AZ was more than 100 times higher than in the SZ, whereas that for syntaxin and SNAP-25 was not significantly different between the AZ and SZ in both the A/C and mossy fiber terminals. These results support the involvement of the t-SNARE proteins in exocytotic fusion in the AZ and the role of CAST in specialization of the membrane domain for the AZ.
Publishing Year
Date Published
2005-08-22
Journal Title
Journal of Comparative Neurology
Volume
489
Issue
2
Page
195 - 216
IST-REx-ID

Cite this

Hagiwara A, Fukazawa Y, Deguchi Tawarada M, Ohtsuka T, Shigemoto R. Differential distribution of release-related proteins in the hippocampal CA3 area as revealed by freeze-fracture replica labeling. Journal of Comparative Neurology. 2005;489(2):195-216. doi:10.1002/cne.20633
Hagiwara, A., Fukazawa, Y., Deguchi Tawarada, M., Ohtsuka, T., & Shigemoto, R. (2005). Differential distribution of release-related proteins in the hippocampal CA3 area as revealed by freeze-fracture replica labeling. Journal of Comparative Neurology, 489(2), 195–216. https://doi.org/10.1002/cne.20633
Hagiwara, Akari, Yugo Fukazawa, Maki Deguchi Tawarada, Toshihisa Ohtsuka, and Ryuichi Shigemoto. “Differential Distribution of Release-Related Proteins in the Hippocampal CA3 Area as Revealed by Freeze-Fracture Replica Labeling.” Journal of Comparative Neurology 489, no. 2 (2005): 195–216. https://doi.org/10.1002/cne.20633.
A. Hagiwara, Y. Fukazawa, M. Deguchi Tawarada, T. Ohtsuka, and R. Shigemoto, “Differential distribution of release-related proteins in the hippocampal CA3 area as revealed by freeze-fracture replica labeling,” Journal of Comparative Neurology, vol. 489, no. 2, pp. 195–216, 2005.
Hagiwara A, Fukazawa Y, Deguchi Tawarada M, Ohtsuka T, Shigemoto R. 2005. Differential distribution of release-related proteins in the hippocampal CA3 area as revealed by freeze-fracture replica labeling. Journal of Comparative Neurology. 489(2), 195–216.
Hagiwara, Akari, et al. “Differential Distribution of Release-Related Proteins in the Hippocampal CA3 Area as Revealed by Freeze-Fracture Replica Labeling.” Journal of Comparative Neurology, vol. 489, no. 2, Wiley-Blackwell, 2005, pp. 195–216, doi:10.1002/cne.20633.

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