牛-山羊异种体细胞克隆胚胎线粒体形态超微结构观察  

The Mitochondrial Ultrastructure of Interspecies Cloned Embryos of Bovine-Goat

作　　者：敬晓棋[1,2] 雷安民[1] 屈雷[2] 窦忠英[1] 

机构地区：[1]西北农林科技大学国家干细胞工程研究中心陕西分中心,陕西杨凌712100 [2]榆林学院生命科学学院生命科学研究中心,陕西榆林719000

出　　处：《中国农业科学》2010年第12期2548-2554,共7页Scientia Agricultura Sinica

基　　金：国家高技术研究发展"863"计划项目(2002AA216161);国家自然科学基金项目(302000137);国家教育部重大项目(03160)

摘　　要：【目的】通过对正常受精胚胎、山羊同克隆胚胎(山羊去核卵与布尔山羊耳皮肤成纤维细胞重组)以及牛-山羊异种克隆胚胎(山羊去核卵与布尔山羊耳皮肤成纤维细胞重组)中线粒体超微结构的观察与比对,从亚细胞水平揭示异种克隆胚胎出现异常的原因,为异种克隆技术的进一步研究提供新思路。【方法】利用透射电镜分别对山羊正常受精胚胎、山羊同种克隆胚胎及牛-山羊异种克隆胚胎的原核、2-细胞、4-细胞、8-细胞及桑椹胚阶段的早期胚胎进行线粒体超微结构观察。【结果】山羊自然受精胚胎、山羊同种克隆胚胎的线粒体均为帽状,随着胚胎的发育,线粒体由电子密度高、嵴少的未成熟状态发育为电子密度低、嵴丰富的成熟线粒体,而牛-山羊异种克隆胚胎从2-细胞胚胎至桑椹胚都存在一种具有多个分叶的线粒体,但这种形态异常的线粒体同样也能够形成电子密度低、嵴丰富的成熟线粒体。【结论】牛卵母细胞与山羊体细胞之间不能有效的进行核-质互作,本研究中具体表现为出现多分叶形线粒体,从而影响牛-山羊异种克隆胚胎的发育。【Objective】 By comparing the mitochondrial ultrastructure among the normal goat embryos,cloned intraspecies goat embryos（reconstructed by goat enucleated oocytes and boer goat ear skin fibroblast） and cloned bovine-goat interspecies embryos（reconstructed by bovine enucleated oocytes and boer goat ear skin fibroblast）,the cause of poor development of cloned interspecies embryos was found at subcellsular level.The result would give a new thinking from different angles to study cloned interspecies embryos.【Method】 The mitochondrial ultrastructure of normal goat embryos,cloned intraspecies goat embryos and cloned bovine-goat interspecies embryos were detected by transmission electron microscope at pronucleus,2-cell,4-cell,8-cell and morulae stages.【Result】 The mitochondria of nomal embryos and cloned intraspecies embryos were cap-like and developed from high electron density and less cristaes to low electron density and rich cristaes with the development of embryo.However,a kind of abnormal-shaped mitochondria of multi-filiation was appeared in the cloned bovine-goat interspecies embryos.These mitochondria could form the normal and rich cristaes,which means they become matured.【Conclusion】 These results showed that the crosstalking between nuclear and cytoplasm could not be regulated effectively,which means that the bovine ooplasm can not reprogamme the goat somatic cell nuclear,and conversely,the goat somatic cell nuclear can not instruct the mitochondria in bovine oocyte efficiently.

关 键 词：牛 山羊 异种克隆 超微结构 线粒体 

分 类 号：S823[农业科学—畜牧学] S827[农业科学—畜牧兽医]