High resolution in situ zymography reveals matrix metalloproteinase activity at glutamatergic synapses
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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MMP activity co-localizes with excitatory synapses.
“Consistent with previous studies in the hippocampus (Wilczynski et al., 2008; Bozdagi et al., 2007; Gawlak et al., 2009), we demonstrated that MMP activity co-localizes with excitatory synapses.”
From Light reintroduction after dark exposure reactivates plasticity in adults via perisynaptic activation of MMP-9 (2017) · cites this paper as Gawlak et al., 2009
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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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