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Miox基因在爪蟾胚胎发育过程中的时间和空间表达谱分析
傅小娟,陈雪梅,周钦,杜晓刚
0
(重庆医科大学第一附属医院肾脏内科;重庆医科大学第一附属医院急诊科;四川大学华西医院生物治疗国家重点实验室遗传工程与小鼠中心)
摘要:
目的 研究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 利用半定量逆转录聚合酶链反应(RT-PCR)观察Miox 在爪蟾胚胎发育过程中的时间表达模式,利用整体原位杂交的方法(WISH)观察Miox基因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,到胚胎发育第28期开始有微量表达,随着胚胎的发育表达量逐渐增高,到胚胎发育第40期Miox表达明显升高,在第41期表达最高,到胚胎第45期表达有所下调。与胚胎第28,34期相比,第40,41,45期表达明显上调(P<0.05);与胚胎第40期相比,第41期表达明显上调(P<0.05);然而,同胚胎第41期相比,第45期表达又明显下调(P<0.05)。WISH发现在30期以前都没能检测到Miox在爪蟾的任何器官有表达,但从33期开始,我们才发现Miox在爪蟾前肾有很微弱的表达, Miox的表达随着发育的进展逐渐增高。同RT-PCR结果相类似,直到第39-40期,Miox的表达量明显升高,并且在随后的时期都以同样高的水平表达。另外我们还发现,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。
关键词:  Miox 爪蟾 肾脏发育 表达模式
DOI:10.3724/SP.J.1008.2014.00832
投稿时间:2014-01-26修订日期:2014-07-10
基金项目:] 国家自然科学基金(81202318,81370816),国家重点基础研究计划(973计划,2011CB944002),重庆自然科学基金一般项目(No.cstc2012jjA10136).Supported by National Natural Science Foundation of China (81202318,81370816), National Key Basic Research Program of China (2011CB944002),Natural Science Foundation Project of CQ CSTC (No.cstc2012jjA10136)[
Identification the Temporal and Spatial Expression Patterns of Miox in the Evolution of Xenopus laevis Embryos
Fu Xiaojuan,Chen Xue-mei,Zhou Qin,Du Xiao-gang
(Department of Nephrology,the First Affiliated Hospital,Chongqing Medical University;Department of Emergency,the First Affiliated Hospital,Chongqing Medical University;Core Facility of Gene Engineered Mice,State Key Laboratory of Biotherapy and Cancer Center,West China Hospital,Sichuan University,Chengdu ,China;Department of Nephrology,the First Affiliated Hospital,Chongqing Medical University;China;;Department of Emergency,the First Affiliated Hospital,Chongqing Medical University;China;;Core Facility of Gene Engineered Mice,State Key Laboratory of Biotherapy and Cancer Center,West China Hospital,Sichuan University,Chengdu ,China)
Abstract:
Objective To identify the molecular evolution of Miox and its temporal and spatial expression patterns in the evolution of Xenopus laevis embryos. Methods The temporal and spatial expression patterns were analyzed by semi-quantitative reverse-transcriptional PCR (RT-PCR) and whole-mount in situ hybridization, respectively. Results We investigated the temporal expression pattern of Miox during embryogenesis of Xenopus laevis by RT-PCR and found that hardly any expression was detected before stage 26. Miox firstly expressed at stage 28 with a very small amount. Its expression level gradually increased along with the development of the process and reaced its peak at stage 40 and kept a high stable level in later period. Compared with stage 28,34,the expression of stage 40,41,45 were upregulated significantly(P<0.05) .Compared with stage 40,the expression of stage 41 was upregulated markedly(P<0.05).But compared with stage 41,the expresssion of stage 45 was downregulated(P<0.05).The results of whole-mount in situ hybridization showed that Miox started express at stage 33 and the expression trend was consistent with RT-PCR. We also found that Miox only expressed in the pronephros tubules in the whole procss of embryo development. Conclusion Miox was tissue-specific in Xenopus laevis pronephros development, which may provide a marker for later pronephros development in the study of organogenesis.
Key words:  Miox Xenopus laevis kidney development expression patterns