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  • Title: SMAD2 overexpression rescues the TGF-β3 null mutant mice cleft palate by increased apoptosis.
    Author: AlMegbel AM, Shuler CF.
    Journal: Differentiation; 2020; 111():60-69. PubMed ID: 31677482.
    Abstract:
    UNLABELLED: During palatal development, medial edge epithelium (MEE) disappearance is one of the crucial steps in the process of fusion. The fate of these cells is still debated, and controversies remain. During secondary palate fusion, TGF-β3 signaling mediated in the cell through the SMAD2 protein plays an important role and leads to the disappearance of the midline epithelial seam (MES) and the confluence of the palatal mesenchyme. In mice, TGF-β3 knock-out is lethal and mice are born with a cleft in the secondary palate. This phenotype has been rescued by targeted overexpression of SMAD2 in the medial edge epithelium (MEE). The goal of this research was to understand the mechanism of palatal fusion in the rescue mice. METHODS: The heads of embryos with four different genotypes (wild-type, K14-SMAD2/TGF-β3(-/-), K14-SMAD2/TGF-β3(±), and TGF-β3 null) were collected at embryonic day E14.5, genotyped, fixed and embedded in paraffin. Serial sections were studied for detection of apoptosis and epithelial mesenchymal transition using immunofluorescence. RESULTS: TGF-β3 null mice developed a cleft in the secondary palate while both mice with K14-SMAD2 overexpression had fusion of the secondary palate. The MEE of both the rescue mice and K14-SMAD2 overexpression had a much higher ratio of apoptotic cells than wild-type mice. The increase in apoptosis was correlated with increased phospho-SMAD2 in the MEE. CONCLUSION: SMAD2 overexpression rescued the cleft in the secondary palate by increasing apoptosis in the medial edge epithelium.
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