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目的研究肌醇加氧酶(myo-inositol oxygenase,Miox)基因在物种进化及爪蟾胚胎发育过程中的时间和空间表达的分布特点。方法利用半定量RT-PCR观察Miox在爪蟾胚胎发育过程中的时间表达模式,利用整体原位杂交方法观察Miox在爪蟾胚胎发育过程中的空间表达模式。结果 RT-PCR结果显示Miox基因在胚胎发育第26期以前都没有表达,至胚胎发育第28期开始有微量表达,随着胚胎的发育其表达量逐渐增高;胚胎发育第40期表达明显升高,第41期时达到最高,第45期时表达有所下降。与胚胎发育第28、34期相比,第40、41、45期表达上调(P<0.05);与胚胎发育第40期相比,第41期表达上调(P<0.05);然而,同胚胎发育第41期相比,第45期表达下调(P<0.05)。整体原位杂交方法发现在胚胎发育30期以前均没能检测到Miox在爪蟾任何器官中的表达,从33期开始,Miox在爪蟾前肾有很微弱的表达,且Miox的表达随着发育的进展逐渐升高。整体原位杂交方法结果同RT-PCR结果相类似,直至第39~40期,Miox的表达才明显升高,并且在随后的时期都以同样高的水平表达。另外,Miox基因在爪蟾胚胎发育过程中均仅仅在原肾小管表达。结论 Miox是一个肾脏特异性基因,对于研究肾脏发育可能提供一个特异性标记。
Objective To study the temporal and spatial distribution of myo-inositol oxygenase (Miox) gene during evolution and Xenopus embryo development. Methods The expression pattern of Miox in Xenopus embryo development was observed by semi-quantitative RT-PCR. The spatial expression patterns of Miox in Xenopus embryo development were observed by in situ hybridization. Results The results of RT-PCR showed that Miox gene was not expressed before the 26th stage of embryo development, and reached the micro level at the 28th stage of embryo development. The expression level of Miox gene gradually increased with the development of embryo and increased significantly at the 40th stage of embryo development , Reaching the highest at the 41st and decreasing at the 45th. Compared with the 28th and the 34th embryos, the 40th, 41th and 45th phases were up-regulated (P <0.05). Compared with the 40th embryos, the 41th phase was up-regulated (P <0.05) Compared with developmental stage 41, the 45th expression was down-regulated (P <0.05). Whole-mount in situ hybridization showed that Miox was undetectable in any organs of Xenopus until 30 days after embryonic development. From the 33rd stage, Miox was weakly expressed in Xenopus laevis, and the expression of Miox was increased with The progress of development is gradually increasing. The results of whole-mount in situ hybridization were similar to those of RT-PCR. Miox expression was not significantly increased until the 39th to 40th cycles, and was expressed at the same high level in subsequent stages. In addition, Miox gene is only expressed in primary renal tubules during Xenopus embryo development. Conclusion Miox is a kidney-specific gene that may provide a specific marker for studying kidney development.