ZFIN ID: ZDB-PUB-110207-33
Differential in vivo zymography: A method for observing matrix metalloproteinase activity in the zebrafish embryo
Keow, J.Y., Herrmann, K.M., Crawford, B.D.
Date: 2011
Source: Matrix biology : journal of the International Society for Matrix Biology   30(3): 169-177 (Journal)
Registered Authors: Crawford, Bryan D.
Keywords: extracellular matrix remodeling, matrix metalloproteinases, in vivo zymography, zebrafish, Xenopus
MeSH Terms:
  • Animals
  • Brain/metabolism
  • Electrophoresis
  • Enzyme Assays/methods*
  • Fluorescence Resonance Energy Transfer
  • Fluorescent Dyes
  • Gene Silencing
  • Matrix Metalloproteinases/genetics
  • Matrix Metalloproteinases/metabolism*
  • Peptides/metabolism
  • Trigeminal Ganglion/metabolism
  • Xenopus laevis/embryology
  • Xenopus laevis/metabolism
  • Zebrafish/embryology
  • Zebrafish/metabolism*
PubMed: 21292002 Full text @ Matrix Biol.
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ABSTRACT
Investigations into the molecular mechanisms of, and cellular signaling pathways modulating ECM remodeling are especially challenging due to the complex post-translational regulation of the primary effectors of ECM catabolism - the matrix metalloproteinases (MMPs). Recently a variety of approaches to the detection of MMP activity have been developed, and the prospect of visualizing ECM remodeling activity in living tissues is now opening exciting avenues of research for matrix biologists. In particular the use of FRET-quenched MMP substrates, which generate a fluorescent signal upon hydrolysis, are becoming increasingly popular, especially because linkers with defined and/or restricted proteolytic sensitivity can be used to bind fluorophore-quencher pairs, making these probes useful in characterizing the activity of specific proteases. We have taken advantage of the transparency and amenability to reverse genetics of the zebrafish embryo, in combination with these fluorogenic MMP substrates, to develop a multiplex in vivo assay for MMP activity that we dub "differential in vivo zymography."
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