目的 :探讨结蛋白对非洲爪蛙胚胎早期发育的影响。方法:利用实时定量RT-PCR和原位杂交方法检测结蛋白在胚胎发育各阶段的表达;通过显微注射特异性反义寡核苷酸进行敲降,利用原位杂交和实时定量RT-PCR方法检测各胚层标志基因表达的变化。结果:结蛋白低水平表达起始于囊胚期,自神经胚期起表达量增加,在随后的发育阶段持续高表达;结蛋白敲降导致胚孔闭合延迟,胚层相关标志基因的表达受抑制。结论:结蛋白自囊胚期起表达于非洲爪蛙胚胎发育各阶段,敲降结蛋白导致胚胎发育迟缓,胚层分化受抑。
Objective:To investigate the effect of desmin on embryo development of Xenopus laevis. Methods:The expression of desmin during embryogenesis was detected by real-time RT-PCR and the whole mount in situ hybridization. The specific antisense oligonucleotides was performed to knockdown the expression of desmin by microinjeetion. The expression of marker genes was tested during germ layer formation by whole mount in situ hybridization and quantitative RT-PCR. Results:The weak expression of desmin starts from the early blastula stage and then detected through all periods of organogenesis. Knockdown of desmin inhibited the blastopore closure,and suppressed the expression of the endodermal,mesodermal and exodermal marker genes. Conclusion:The expression of desmin starts from the blastula during embryogenesis of Xenopus lavies. Loss function of desmin leads to embryonic growth retardation and depressed germ-layer formation.