Sasaki Yasunori, O'Kane Sharon, Dixon Jill, Dixon Michael J, Ferguson Mark W J
Faculty of Life Sciences, University of Manchester, Manchester, UK.
Arch Oral Biol. 2007 Mar;52(3):260-7. doi: 10.1016/j.archoralbio.2006.09.012. Epub 2006 Nov 13.
Transforming growth factor-beta (TGF-beta3) gene disruption causes cleft secondary palate. Pax9 and Sonic hedgehog (Shh) genes are involved in the patterning of vertebrate embryonic tissues, including the facial skeleton. We investigated the expression of Pax9 and Shh genes during normal mouse palate development and in the developing cleft palates of TGF-beta3 null embryos. Whole mount in situ hybridization was conducted with use of Pax9 and Shh riboprobes for TGF-beta3 null, heterozygous and wild type mice at E12.5-E16.5. Histological analysis was processed by section in situ hybridization. In the wild type, Pax9 and Shh were expressed in the palate between E12.5-E15.5. Shh expression in the secondary palate was restricted to the rugae and the soft palate. Pax9 expression was predominantly in the palatal medial edge between E14.5 and E15.5. These patterns suggest that Shh and Pax9 may have different functions during palate development. In TGF-beta3 null mice, both genes expression patterns in the palate were different to those in wild type mice. In TGF-beta3 null mice, Pax9 expression was much reduced in the palatal medial edge at the critical time of palatal fusion (E14.5-E15.5). Shh expression in the palates of TGF-beta3 null mice was reduced throughout E12.5-E15.5, whilst Shh expression in heterozygous did not appear down regulated compared with the wild type. These results indicate that Pax9 and Shh expression are altered when the TGF-beta3 gene is deleted and suggest that Pax9 and Shh may be involved in the TGF-beta3 regulation of normal palatal fusion.
转化生长因子-β（TGF-β3）基因缺失会导致腭裂。Pax9和音猬因子（Shh）基因参与脊椎动物胚胎组织（包括面部骨骼）的模式形成。我们研究了Pax9和Shh基因在正常小鼠腭部发育过程中以及TGF-β3基因敲除胚胎发育中的腭裂中的表达情况。使用Pax9和Shh核糖探针，对E12.5 - E16.5的TGF-β3基因敲除、杂合子和野生型小鼠进行了全胚胎原位杂交。通过切片原位杂交进行组织学分析。在野生型中，E12.5 - E15.5期间腭部有Pax9和Shh表达。Shh在继发腭中的表达局限于腭皱襞和软腭。Pax9表达主要在E14.5和E15.5之间的腭内侧边缘。这些模式表明，Shh和Pax9在腭部发育过程中可能具有不同功能。在TGF-β3基因敲除小鼠中，腭部这两个基因的表达模式与野生型小鼠不同。在TGF-β3基因敲除小鼠中，在腭融合的关键时期（E14.5 - E15.5），腭内侧边缘的Pax9表达大幅减少。TGF-β3基因敲除小鼠腭部的Shh表达在整个E12.5 - E15.5期间均减少，而杂合子中的Shh表达与野生型相比未出现下调。这些结果表明，当TGF-β3基因缺失时，Pax9和Shh的表达会发生改变，提示Pax9和Shh可能参与TGF-β3对正常腭融合的调控。
