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Type: Artigo de periódico
Title: In Situ Zymography and Immunolabeling in Fixed and Decalcified Craniofacial Tissues
Author: Porto, IM
Rocha, LB
Rossi, MA
Gerlach, RF
Abstract: In situ zymography is a very important technique that shows the proteolytic activity in sections and allows researchers to observe the specific sites of proteolysis in tissues or cells. it is normally performed in non-fixed frozen sections and is not routinely performed in calcified tissues. In this study, we describe a technique that maintains proteolytic activity in fixed and decalcified sections obtained after routine paraffin sectioning in conventional microtome and cryostat sections. We used adult rat hemimandibles, which presented bone, enamel, and dentine matrices; the substrate used was dye-quenched-gelatin. Gelatinolytic activity was colocalized with MMP-2 using fluorescent antibodies. Specific proteolytic activity was observed in all sections, compatible with metalloproteinase activity, particularly in dentine and bone. Furthermore, matrix metalloproteinase-2 was colocalized to the sites of green fluorescence in dentine. In conclusion, the technique presented here will allow in situ zymography reactions in fixed, decalcified, and paraffin-embedded tissues, and we showed that paraformaldehyde-lysine-periodate-fixed cryostat sections are suitable for colocalization of gelatinolytic activity and protein labeling with antibodies. (J Histochem Cytochem 57:615-622, 2009)
Subject: craniofacial tissues
in situ zymography
proteolytic activity
fluorescence microscopy
matrix metalloproteinase-2
Country: EUA
Editor: Histochemical Soc Inc
Citation: Journal Of Histochemistry & Cytochemistry. Histochemical Soc Inc, v. 57, n. 7, n. 615, n. 622, 2009.
Rights: fechado
Identifier DOI: 10.1369/jhc.2009.952127
Date Issue: 2009
Appears in Collections:Unicamp - Artigos e Outros Documentos

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