Development of secondary palate requires strict regulation of ECM remodeling: sequential distribution of RECK, MMP-2, MMP-3, and MMP-9.
We have evaluated RECK (reversion-inducing-cysteine-rich protein with Kazal motifs), MMP-2 (matrix metalloproteinase-2), MMP-3, and MMP-9 involvement during palate development in mice by using various techniques. Immunohistochemical features revealed the distribution of RECK, MMP-2, and MMP-3 in the mesenchymal tissue and in the midline epithelial seam at embryonic day 13 (E13), MMPs-2, -3, and -9 being particularly expressed at E14 and E14.5. In contrast, RECK was weakly immunostained at these times. Involvement of MMPs was validated by measuring not only their protein expression, but also their activity (zymograms). In situ hybridization signal (ISH) for RECK transcript was distributed in mesenchymal and epithelial regions within palatal shelves at all periods evaluated. Importantly, the results from ISH analysis were in accord with those obtained by real-time polymerase chain reaction. The expression of RECK was found to be temporally regulated, which suggested possible roles in palatal ontogeny. Taken together, our results clearly show that remodeling of the extracellular matrix is finely modulated during secondary palate development and occurs in a sequential manner.
de Oliveira Demarchi
Ana Claudia Cardoso
AC
Department of Biochemistry, IB, University of Campinas, Campinas, SP, Brazil.
Zambuzzi
Willian Fernando
WF
Paiva
Katiúcia Batista Silva
KB
da Silva-Valenzuela
Maria das Graças
Md
Nunes
Fabio Daumas
FD
de Cássia Sávio Figueira
Rita
R
Sasahara
Regina Maki
RM
Demasi
Marcos Angelo Almeida
MA
Winnischofer
Sheila Maria Brochado
SM
Sogayar
Mari Cleide
MC
Granjeiro
José Mauro
JM
D000818:Animals; D019521:Body Patterning / Q000502:physiology; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D058851:GPI-Linked Proteins; D018507:Gene Expression Regulation, Developmental / Q000502:physiology; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism*; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism*; D045726:Metalloproteases / Q000378:metabolism*; D051379:Mice; D010159:Palate / Q000166:cytology / Q000196:embryology* / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction
D016326:Extracellular Matrix Proteins; D058851:GPI-Linked Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; C492025:Reck protein, mouse; D045726:Metalloproteases; D019278:Matrix Metalloproteinase 3; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9