Miox基因在爪蟾胎发育中的时间和空间表达谱分析.docVIP

Miox基因在爪蟾胚胎发育中的时间和空间表达谱分析 傅小娟1，陈雪梅2，周钦3，杜晓刚* 重庆医科大学第一附属医院肾脏内科，重庆400016 重庆医科大学第一附属医院急诊科，重庆 400016 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，成都 610000 ［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2834期相比，第4041、,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， 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。WISH同RT-PCR结果相类似，直到第3940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 ［中图分类号］ R 394.1 ［文献标识码］ A Identification the Temporal and Spatial Expression Patterns of iox in the Evolution of Xenopus laevis Embryos FU Xiao-juan1，C Xue-mei2，Z Qin3,DU Xiao-gang1* 1. Department of Nephrology, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China 2. Department of Emergency, the First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China 3. Core Facility of Gene Engineered Mice, State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, Sichuan University, Chengdu 610000,Sichuan,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 sta