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原文摘要：; w7 m3 E; u+ a2 f6 s& ~
2 s/ U9 D. Q$ x! h: w W9 wProteome of mouse oocytes at different developmental stages5 W5 x( E* I+ m: q! u- e. g
" N$ a. w/ D' C/ Y* |The mammalian oocyte possesses powerful reprogramming factors, which can reprogram terminally differentiated germ cells (sperm) or somatic cells within a few cell cycles. Although it has been suggested that use of oocyte-derived transcripts may enhance the generation of induced pluripotent stem cells, the reprogramming factors in oocytes are undetermined, and even the identified proteins composition of oocytes is very limited. In the present study, 7,000 mouse oocytes at different developmental stages, including the germinal vesicle stage, the metaphase II (MII) stage, and the fertilized oocytes (zygotes), were collected. We successfully identified 2,781 proteins present in germinal vesicle oocytes, 2,973 proteins in MII oocytes, and 2,082 proteins in zygotes through semiquantitative MS analysis. Furthermore, the results of the bioinformatics analysis indicated that different protein compositions are correlated with oocyte characteristics at different developmental stages. For example, specific transcription factors and chromatin remodeling factors are more abundant in MII oocytes, which may be crucial for the epigenetic reprogramming of sperm or somatic nuclei. These results provided important knowledge to better understand the molecular mechanisms in early development and may improve the generation of induced pluripotent stem cells.
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作者简介：) O6 L9 `& \0 F1 f* @2 K, t
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高绍荣 博士3 J7 n0 s# V6 k* j/ S& p
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北京生命科学研究所研究员/ m0 c! |) a0 V. H: e
+ v/ g( i9 l+ i% g; o$ o研究概述： x5 J0 U1 ]5 ?4 z( a1 {
2 k( l. }* J2 P d5 h本实验室工作主要集中在研究哺乳动物体细胞克隆胚胎发育过程中再编程(Reprogramming)的分子机理。将体细胞核通过显微操作移入去除遗传物质的成熟卵母细胞中从而启动胚胎发育是一个由体细胞基因表达逆转进入胚胎细胞基因表达的再编程过程，正是由于我们对卵母细胞再编程的分子机理不清楚才使克隆成功的效率仍然处于非常低的水平。而能够分离纯化卵母细胞中特有的再编程因子(Reprogramming Factors) 将会对克隆效率的提高以及干细胞的研究起到十分重要的意义。我们今后工作重点之一就是要通过核移植并结合生物化学的方法鉴定卵母细胞胞质中的这些重要因子。
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) g% n+ V, Z' E# l5 P近来，人们已经意识到胚胎干细胞的研究对一些疾病治疗的重大前景。通过核移植的方法可以得到与病人遗传信息相同的胚胎干细胞，从而可以消除移植后免疫排斥的可能。我们实验室已经通过核移植的方法建立起了数十株小鼠体细胞克隆胚胎干细胞系并正在进行体外分化为心肌组织的研究和体内移植后功能的鉴定。同时我们正在进行人治疗性克隆的研究以期尽快建立病人来源的体细胞核移植胚胎干细胞系。, P8 z$ u9 z: ?6 ]2 r+ }7 |( \; }2 q) F
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小鼠胚胎发育过程中卵母细胞与早期胚胎极性发生机理的研究也是我们实验室的重要研究方向。5 K& z" u; f, q a! H8 \
3 H7 k4 m) P5 e2 v! L% T因此，本实验室以后的工作主要集中在：
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1. 研究克隆胚胎再编程(Reprogramming)的分子机理，鉴定与纯化卵母细胞中特有的再编程因子(Reprogramming factor)。
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2. 建立克隆胚胎干细胞系并进行体外分化与功能的研究。
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9 X+ e7 D& n- b/ C1 J' J9 N* G( t: K3. 小鼠卵母细胞与早期胚胎极性发生机理的研究。 |
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发表于 2010-9-30 20:59
发表于 2010-9-30 21:21
发表于 2010-10-1 14:05
