Differential distribution of release‐related proteins in the hippocampal CA3 area as revealed by freeze‐fracture replica labeling
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 (A/C) 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 mos…
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The mossy fiber-CA3 PN synapse is accessible to quantitative structural analysis by electron microscopy, high-pressure freezing, and freeze-fracture replica labeling.
“Finally, it is accessible to quantitative structural analysis by EM [34,35], high-pressure freezing (HPF) [36–38], and freeze-fracture replica labeling (FRL) [39].”
From Presynaptic cAMP-PKA-mediated potentiation induces reconfiguration of synaptic vesicle pools and channel-vesicle coupling at hippocampal mossy fiber boutons (2024) · cites this paper as [39]
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Self-citationDYNAMIQUE DE L'ASSEMBLAGE MOLÉCULAIRE SYNAPTIQUE : ÉTUDE DE LA DIFFUSION LATÉRALE DE LA SYNTAXIN 1A
Ribrault, C. (2010) · HAL
- Authors
- Hagiwara, A., Fukazawa, Y., Deguchi‐Tawarada, M., Ohtsuka, T., & Shigemoto, R.
- Affiliations
- The Graduate University for Advanced Studies, SOKENDAI (JP)
- KAN Research Institute (JP)
- Corresponding author
- Hagiwara, Akari
- Venue
- Journal of Comparative Neurology
- Publisher
- Wiley
- Published
- 2005-06-27
- Volume
- 489
- Issue
- 2
- Pages
- 195-216
- ISSN
- 0021-9967 (ISSN-L)
- 1096-9861
- OpenAlex
- W2024791462
- Cited by
- 90
- References
- 86
- Citations by year
- 2024: 2 · 2023: 4 · 2022: 6 · 2021: 5 · 2020: 3 · 2019: 1 · 2018: 5 · 2017: 4 · 2016: 5 · 2015: 3 · 2014: 5 · 2013: 3 · 2012: 7
- FWCI
- 2.98
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- The Journal of Comparative Neurology (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
- Keywords
- active zone, synaptic vesicle release, syntaxin, SNAP-25, CAST, bassoon, hippocampal CA3, mossy fiber terminals
- MeSH terms
- Animals, Freeze Fracturing, Hippocampus, Immunohistochemistry, Male, Membrane Proteins, Nerve Tissue Proteins, Microscopy, Immunoelectron, Rats, Sprague-Dawley, Presynaptic Terminals, Receptors, Metabotropic Glutamate, Vesicular Transport Proteins, SNARE Proteins, Qa-SNARE Proteins, Synaptosomal-Associated Protein 25, Rats
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