The Action Mechanism Of Brachyury In Mouse Embryonic Development And Bone Marrow Mesenchynal Stem Cells (BMSCs)

The Erachyury gene is essential for the const ruction of nouse enbryonic nesoderm,and the heterozygous nice with Brachyury gene notation show a short tail phennotype.BMP and FGF signaling pathways can control physiological processes such as embryo fornation and bone development in nouse enbryonic development and bone narrownessenchynal stemoells(EMSCs)The interaction of Brachyuy genes with BMP and FGF signaling pathways may cause short tail Generation of phenotype.In order to reveal the effect of Erachyuy gene notation on the BMP signal pathway during the development of nouse enbryos,the RNA of 7.5dpc~12.5dpc wild-type and not ant mouse enbryos was first extract ed and analyzed by RT-cPCR Expression of Brachyury Gene and Related Genes in BMP Pat hway in Two Model Mouse Enbryos:(1)Brachyury's expression in nornal wild-type and Brachyury not ant mouse enbryos is consistent,reading a peak of 9.5 dpc,however,the expression level of the wild type is higher than that of the not ant type.(2)The expression of Smad1,Smad5,Smad8,BMP2,BMP4,BMP7,RUNX2 and SOX9 did not change significantly from7.5dpc 12.5dpc in wild-type embryos,but Smad1,Smad5,Smad8,BMP2 and The expression of SOX9 was highest at 9.5 dpc.Among them,the expression of SOX9 at 9.5dpc was much higher than other genes The peaks of BMP4 and RUNX2 appeared at 11.5 dpc,but the change of BMP7 expression was not much different from that of wild type.We sought to investigate the potential interaction between Brachyury gene and BMP and FGF signaling pathway in bone narrowmesenchynal stemoells(EMSCs).In this experiment,BMSCs of Brachyury gene natation model mice were oultured by whole bone narrow culture method.Different oonoentrations of FGFR3 inhibitors were added to the cells,and the effects on Brachyury gene and EMP signaling pathways and FGF signaling pathway-related factors were detected.The experi mental results are as follows:(1)Brachyury expresses higher levels of mutant BMSCs with different concentrations of FGFR3 inhibitors than BMSCs without inhibitors,and Brachyury expresses the highest at 25?M Snad1,Snad5,Snad8,and SOX9 have the same trend as Brachyury,(2)In mut ant BMSCs,the FGFR3 inhibitor ooncertration in the range of 5~15?M has a oertain inhibitory effect on FGFR1 and FGFR2,but the strongest inhibitory ooncentration on FGFR3 is 1 ?M and 5?MIt is not difficult to see from the experimental results hat Brachyury,as a transcription factor,has a certain regulatory effect on the expression of two signaling pathway components This research conbined with the team's previous research contents revealed the potential relationship between Brachyury and related componerts of the BMP and FGF signaling pathways.At the genetic level,we studied some mechanisns of Brachyury genes in mouse enbryonic development and BMSCs.