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inositol-requiring enzyme 1α is required for gut development in Xenopus lavies embryos

World Journal of Gastroenterology 2013年第2期19卷 227-234页

作者： Jing Guo Xin-Xin Li Jiao-Jiao Feng Chen-Yang Yin XueJun Wang Ning Wang Li Yuan Department of Biochemistry and Molecular Biology Nanjing Medical University

AIM:To investigate the role of inositol-requiring enzyme 1α(IRE1α) in gut development of Xenopus lavies ***:Xenopus embryos were obtained with in vitro fertilization and cultured in 0.1 × *** and half nanogram of IRE1α,1 ng of IRE1α-GR mRNA,1 ng of IRE1αΔC-GR mRNA,and 50 ng of IRE1α morpholino oligonucleotide(MO) or XBP1(C)MO were injected into four blastomeres at 4-cell stage for scoring the phenotype and marker gene *** rescue the effect of IRE1α MO,1 ng of IRE1α-GR mRNA was coinjected with 50 ng of *** the activation of the GR-fusion proteins,dexamethasone was prepared as 5 mmol/L stock solutions in 100% ethanol and applied to the mRNA injected embryos at desired stages in a concentration of 10 μmol/L in 0.1 × *** were kept in dexamethasone up to stage ***-mount in situ hybridization was used to determine specific gene expression,such as IRE1α,IRE1β,Xbra and Xsox17α.IRE1α protein expression during Xenopus embryogenesis was detected by Western ***:In the whole-mount in situ hybridization analysis,xenopus IRE1α and IRE1β showed quite different expression pattern during tadpole *** relatively higher expression of IRE1α was observed in the pancreas,and significant transcription of IRE1β was found in the ***1α protein could be detected at all developmental stages analyzed,from stage 1 to stage ***-of-function assay showed that IRE1α mRNA injected embryos at tailbud stage were nearly normal and the expression of the pan-mesodermal marker gene Xbra and the endodermal gene Xsox17α at stage 10.5 was not significantly changed in embryos injected with IRE1α mRNA as compared to uninjected control *** at tadpole stage,the embryos injected with IRE1α-GR mRNA did not display overt phenotype,such as gut-coiling ***-of-function assay demonstrated that the IRE1α MO injected embryos were morphologically normal before the tailbud *** did not observe a significant change of mesodermal and end

关键词： inositol-requiring enzyme 1α XBP1 Xenopus lavies Gut Development

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