目的:探讨X盒结合蛋白1(X-box binding protein 1,XBP1)对非洲爪蟾胚胎发育的影响。方法:利用Western blot、原位杂交和免疫染色法检测XBP1在胚胎发育各个阶段的表达;通过显微注射基因特异性反义寡核苷酸实现基因敲降,利用6T- PC6和整体胚胎原位杂交法检测基因表达变化。结果:XBP1主要表达于非洲爪蟾胚胎发育期的神经、前肾、胰腺等器官中;敲降XBP1s在胚胎发育早期影响非洲爪蟾胚层形成相关基因,在胚胎发育后期影响胰腺发育相关因。结论:XBP1s在胚胎发育早期可能影响胚层形成,在晚期可能影响胰腺的发育。
Abstract:
Objective:To investigate the effect of X-box binding protein 1 (XBP1) on embryogenesis of Xenopus laevis. Methods: The expression of XBP1 during each stage of embryogenesis was detected by Western blotting assay, the whole mount in situ hybridization,and immunostaining. The micro-injection of morpholino (MO) was performed to knockdown XBP1. The expression of marker genes was tested during germ layer formation,as well as embryo development by whole mount in situ hybridization and real-time PCR. Results: XBP1 expressions of the nerve,anterior kidney,pancreas,and other organs were detected during embryogenesis of Xenopus laevis. Knockdown of XBP1 inhibited the expression of germ layer related genes at the early stage of embryogenesis,and then inhibited pancreas development related gene in Xenopus laevis embryos at the late stage of embryogenesis. Conclusion :XBP1 may be required for the germ layer formation at the early stage of embryogenesis and pancreas development at the late stage of embryogenesis.