Cited by 101Protease and Inhibitor MechanismsNeuroscience and Neuropharmacology ResearchNeuroinflammation and Neurodegeneration Mechanisms

High resolution in situ zymography reveals matrix metalloproteinase activity at glutamatergic synapses

Gawlak, M., Górkiewicz, T., Gorlewicz, A., Konopacki, F.A., Kaczmarek, L., Wilczynski, G.M.·2009·Neuroscience

Abstract

Synaptic plasticity involves remodeling of extracellular matrix. This is mediated, in part, by enzymes of the matrix metalloproteinase (MMP) family, in particular by gelatinase MMP-9. Accordingly, there is a need of developing methods to visualize gelatinolytic activity at the level of individual synapses, especially in the context of neurotransmitters receptors. Here we present a high-resolution fluorescent in situ zymography (ISZ), performed in thin sections of the alcohol-fixed and polyester wax-embedded brain tissue of the rat (Rattus norvegicus), which is superior to the current ISZ protocols. The method allows visualization of structural details up to the resolution-limit of light microscopy, in conjunction with immunofluorescent labeling. We used this technique to visualize and quantify gelatinolytic activity at the synapses in control and seizure-affected rat brain. In particular, we demonstrated, for the first time, frequent colocalization of gelatinase(s) with synaptic N-methyl-D-aspartic acid (NMDA)- and AMPA-type glutamate receptors. We believe that our method represents a valuable tool to study extracellular proteolytic processes at the synapses, it could be used, as well, to investigate proteinase involvement in a range of physiological and pathological phenomena in the nervous system.

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Crossref · 10.1016/j.neuroscience.2008.05.045

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Authors
Gawlak, M., Górkiewicz, T., Gorlewicz, A., Konopacki, F. A., Kaczmarek, L., & Wilczynski, G. M.
Affiliations
Instytut Biologii Doświadczalnej im. Marcelego Nenckiego (PL)
Corresponding author
  • Kaczmarek, L.
  • Wilczynski, G.M.
Venue
Neuroscience
Publisher
Elsevier BV
Published
2009-01
Volume
158
Issue
1
Pages
167-176
ISSN
0306-4522 (ISSN-L)
OpenAlex
W1989777389
Cited by
101
References
32
Citations by year
2026: 3 · 2025: 2 · 2024: 3 · 2023: 4 · 2022: 4 · 2021: 5 · 2020: 5 · 2019: 4 · 2018: 3 · 2017: 5 · 2016: 7 · 2015: 9 · 2014: 6 · 2013: 8 · 2012: 13
FWCI
2.26
Type
journal article
Language
en
Open access
closed
Locations
  • Neuroscience (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Protease and Inhibitor Mechanisms
  • Life Sciences › Neuroscience › Cellular and Molecular Neuroscience › Neuroscience and Neuropharmacology Research
  • Life Sciences › Neuroscience › Neurology › Neuroinflammation and Neurodegeneration Mechanisms
Keywords
matrix metalloproteinases, MMP-9, gelatinase activity, glutamatergic synapses, synaptic plasticity, extracellular matrix remodeling, NMDA receptor, AMPA receptors, seizures, rat brain
MeSH terms
Animals, Biological Assay, Brain, Epilepsy, Extracellular Matrix, Fluorescent Antibody Technique, Male, Microscopy, Fluorescence, Neurochemistry, Synapses, Receptors, N-Methyl-D-Aspartate, Tissue Embedding, Tissue Fixation, Rats, Wistar, Receptors, AMPA, Glutamic Acid, Matrix Metalloproteinases, Proteomics, Rats

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