High resolution in situ zymography reveals matrix metalloproteinase activity at glutamatergic synapses
| Autor: | Grzegorz M. Wilczynski, Maciej Gawlak, Adam Gorlewicz, T. Górkiewicz, Filip A. Konopacki, Leszek Kaczmarek |
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| Rok vydání: | 2009 |
| Předmět: |
Male
Proteomics Nervous system Tissue Fixation Fluorescent Antibody Technique Glutamic Acid Biology Receptors N-Methyl-D-Aspartate Synapse Extracellular medicine Animals Gelatinase Receptors AMPA Rats Wistar Epilepsy Tissue Embedding General Neuroscience Glutamate receptor Brain Colocalization Neurochemistry Long-term potentiation Matrix Metalloproteinases Extracellular Matrix Rats Cell biology medicine.anatomical_structure Microscopy Fluorescence Biochemistry Synapses Synaptic plasticity Biological Assay |
| Zdroj: | Neuroscience. 158:167-176 |
| ISSN: | 0306-4522 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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