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体内酶谱分析:研究特定脑细胞中基质金属蛋白酶活性的重要活性检测方法。

In Vivo Zymography: An Essential Activity Assay for Studying the Activity of Matrix Metalloproteinases in Specific Brain Cells.

作者信息

Hodebourg Ritchy

机构信息

Medical University of South Carolina, Charleston, SC, USA.

出版信息

Methods Mol Biol. 2025;2918:21-33. doi: 10.1007/978-1-0716-4482-9_3.

Abstract

Matrix metalloproteinases (MMPs) are a group of zinc endopeptidases that break down the extracellular matrix. MMP-2 and MMP-9 are particularly abundant in the brain and are essential for synaptic plasticity. When studying synaptic plasticity, it is crucial to measure their activity in both normal and pathological conditions. Although gel and in situ zymography are commonly used to assess MMP proteolytic activity, these methods are typically applied to samples and tissues ex vivo and do not allow the quantification of MMP function during a behavioral experiment. Consequently, the in vivo zymography represents the ideal activity assay to study the MMP function. Moreover, we recently developed a method to quantify MMP activity in specific cell types in rat brains. This chapter details the in vivo zymography approach used to accurately quantify the gelatinolytic activity of MMP-2 and MMP-9 in rat brains in both non-cell-specific and cell-specific manners.

摘要

基质金属蛋白酶(MMPs)是一组能分解细胞外基质的锌内肽酶。MMP-2和MMP-9在大脑中含量尤为丰富,对突触可塑性至关重要。在研究突触可塑性时,在正常和病理条件下测量它们的活性至关重要。尽管凝胶和原位酶谱法通常用于评估MMP的蛋白水解活性,但这些方法通常应用于离体样本和组织,无法在行为实验中对MMP功能进行定量。因此,体内酶谱法是研究MMP功能的理想活性测定方法。此外,我们最近开发了一种方法来定量大鼠脑中特定细胞类型的MMP活性。本章详细介绍了用于以非细胞特异性和细胞特异性方式准确量化大鼠脑中MMP-2和MMP-9明胶酶解活性的体内酶谱法。

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