title: Development of secondary palate requires strict regulation of ECM remodeling: sequential distribution of RECK, MMP-2, MMP-3, and MMP-9.
abstract: 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.
authors.affiliation: Department of Biochemistry, IB, University of Campinas, Campinas, SP, Brazil.
authors.forename: Ana Claudia Cardoso
authors.initials: AC
authors.lastname: de Oliveira Demarchi
authors.forename: Willian Fernando
authors.initials: WF
authors.lastname: Zambuzzi
authors.forename: Katiúcia Batista Silva
authors.initials: KB
authors.lastname: Paiva
authors.forename: Maria das Graças
authors.initials: Md
authors.lastname: da Silva-Valenzuela
authors.forename: Fabio Daumas
authors.initials: FD
authors.lastname: Nunes
authors.forename: Rita
authors.initials: R
authors.lastname: de Cássia Sávio Figueira
authors.forename: Regina Maki
authors.initials: RM
authors.lastname: Sasahara
authors.forename: Marcos Angelo Almeida
authors.initials: MA
authors.lastname: Demasi
authors.forename: Sheila Maria Brochado
authors.initials: SM
authors.lastname: Winnischofer
authors.forename: Mari Cleide
authors.initials: MC
authors.lastname: Sogayar
authors.forename: José Mauro
authors.initials: JM
authors.lastname: Granjeiro
mesh_terms: 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
chemical_list: 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