An Msx2 -Sp6-Follistatin Pathway Operates During Late Stages of Tooth Development to Control Amelogenesis

Frontiers in Physiology
Intan RuspitaMarianna Bei

Abstract

Ameloblasts are epithelially derived cells responsible for enamel formation through a process known as amelogenesis. Amongst the several transcription factors that are expressed during amelogenesis, both Msx2 and Sp6 transcription factors play important role. Msx2 and Sp6 mouse mutants, exhibit similar amelogenesis defects, namely enamel hypoplasia, while humans with amelogenesis imperfecta (AI) carry mutations in the human homologues of MSX2 or SP6 genes. These across species similarities in function indicate that these two transcription factors may reside in the same developmental pathway. In this paper, we test whether they work in a coordinated manner to exert their effect during amelogenesis. Two different dental epithelial cell lines, the mouse LS8 and the rat G5 were used for either overexpression or silencing of Msx2 or Sp6 or both. Msx2 mutant mouse embryos or pups were used for in vivo studies. In situ hybridization, semi-quantitative and quantitative real time PCR were employed to study gene expression pattern. MatInspector was used to identify several potential putative Msx2 binding sites upstream of the murine Sp6 promoter region. Chromatin Immunoprecipitation (chIP) was used to confirm the binding of Msx2 to Sp6 p...Continue Reading

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Methods Mentioned

BETA
transgenic
transfection
gene knock down
PCR
Immunoprecipitation
Assay
chIP
reverse transcription PCR
gene knockdown

Software Mentioned

Primer
GraphPad prism
UCSC MatInspector
ImageJ

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