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Mouse round spermatids were electrofused with homologous mature oocytes to examine the be-haviour of their nuclei within the ooplasm and the abilities of development. A single spermatid was injected in the perivitelline space of a mature oocyte and an electron fusion pulse was given. The best round spermatid-oocyte pairs (RS-O) fusion took place at 20-30 s AC (1 MHz, 50V/cm) followed by a single fusion DC pulse (3 700-3 800 V/cm, 25 μs) and another 30 s AC current. The total survival rate and fusion rate of RS-O were 89.0% (575/646) and 61. 9% (356/575), respectively. 49.2% (175/356) of fused oocytes developed to 2PN stage . The concentration of Ca2 in the fusion medium produced no significant effect on the above targets. The 2PN development rate of the fused RS-O from the oocytes collected 14-16 h after hCG injection was higher than others. 32.6% (57/175) of the 2PN oocytes had fully developed spermatid (male) and oocyte (female) pronuclei. The rest spermatid-derived pronu-clei remained small in size 相似文献
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大熊猫供核体细胞在兔卵胞质中可去分化而支持早期重构胚发育 总被引:26,自引:0,他引:26
体外培养的成年大熊猫骨骼肌细胞、子宫上皮细胞和乳腺细胞 ,分别作为供核细胞移植进入去核兔卵母细胞中以构建异种重构胚 .3种组织来源的体细胞在去核兔卵质中均可去分化、恢复全能性和替代合子核进行卵裂 ,并支持早期重构胚发育 .其中以乳腺细胞效果最好 ,子官上皮细胞次之 ,骨骼肌细胞最差 .对比实验表明 ,胞质直接注射法结合重构卵在体培养可比电融合法加体外培养方案获得更大比例的囊胚率 .染色体分析表明异种重构胚的核遗传物质来自大熊猫供体体细胞核 .线粒体DNA分析表明重构囊胚中存在有大熊猫线粒体 .这些结果初步说明 :( 1 )卵胞质使体细胞核去分化不具种特异性 ;( 2 )哺乳动物异种重构胚早期发育中 ,异种细胞核与细胞质之间是相容的 . 相似文献
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钙调素(CaM)拮抗剂三氟拉嗪(TFP)在10~100 μmol/L浓度范围内对豚鼠精子获能和顶体反应具有极显著的促进作用,且不依赖于外源Ca2+的存在.Forskolin,dbcAMP和咖啡因对非同步培养系统获能早期豚鼠精子的顶体反应有显著的促进作用,但对同步培养系统获能早期精子的顶体反应无显著影响.若在同步培养系统获能培养较长时间(15h)使精子获能,则这3种药物对顶体反应均具有显著的促进作用,且不依赖外源Ca2+的存在.蛋白激酶C(PKC)激活剂PMA和PDB对获能前期豚鼠精子的顶体反应无显著影响,但对获能精子的顶体反应有显著的促进作用,且这种促进作用也不依赖于外源Ca2+的存在.相反,PKC抑制剂stau-rosporine对顶体反应有显著的抑制作用. 相似文献
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MSH2 7是一株抗小鼠精子的单克隆抗体 ,此抗体及其纯化的抗原分子在体外受精实验中均可以阻止精卵膜融合 ,因而推测其抗原在精卵膜融合过程中具有重要的作用 .这里探讨了抗原分子在精子睾丸后成熟过程中MSH2 7识别表位的显露条件 .MSH2 7抗原分子在睾丸中产生 ,定位于精子顶体后区的质膜上 ;但MSH2 7所识别的抗原表位只在顶体反应后的精子上表露 .在受精过程中 ,精子必须完成顶体反应并穿过卵子的透明带才能接近卵子质膜并与之融合 .同时还研究了顶体反应和穿透明带作用对MSH2 7表位显露的影响 .在附睾成熟过程中 ,随着精子在附睾中的转运 ,它们发生顶体反应的能力不断增加 ;精子抗原表位的显露呈现顶体反应依赖性 ,且两者呈现线性回归关系 .钙离子载体A2 3 187诱发顶体反应后 ,MSH2 7染色阳性的精子远远少于顶体反应的精子 ;但穿过透明带后绝大多数精子 ( 98% )呈现MSH2 7染色阳性 .总之 ,在睾丸后成熟 (附睾成熟、获能、顶体反应 )和精子穿透明带的过程中 ,对整个精子群体来讲 ,单抗MSH2 7所识别的抗原表位的显露和精子最终获得穿入卵子质膜的能力是同步发生的 . 相似文献
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The giant panda skeletal muscle cells, uterus epithelial cells and mammary gland cells from an adult individual were cultured and used as nucleus donor for the construction of interspecies embryos by transferring them into enucleated rabbit eggs. All the three kinds of somatic cells were able to reprogram in rabbit ooplasm and support early embryo development, of which mammary gland cells were proven to be the best, followed by uterus epithelial cells and skeletal muscle cells. The experiments showed that direct injection of mammary gland cell into enucleated rabbit ooplasm, combined with in vivo development in ligated rabbit oviduct, achieved higher blastoeyst development than in vitro culture after the somatic cell was injected into the perivitelline space and fused with the enucleated egg by electrical stimulation. The chromosome analysis demonstrated that the genetic materials in reconstructed blastocyst cells were the same as that in panda somatic cells. In addition, giant panda mitochondrial DNA ( 相似文献
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利用杂交瘤技术制备小鼠抗精子单克隆抗体 6B1 0 ,为IgG型免疫球蛋白 .通过 (NH4) 2 SO4盐析并结合蛋白A SepharoseCL 4B亲和层析法纯化单克隆抗体6B1 0 ,用SDS 聚丙烯酰胺凝胶电泳分析抗体纯度 .采用金霉素荧光染色法 (CTC)检测精子顶体反应 ,发现单克隆抗体 6B1 0可以显著地抑制孕酮诱导的精子顶体反应 .采用间接免疫荧光法和胶体金免疫电镜观察发现单克隆抗体 6B1 0的相应抗原定位于精子顶体质膜上 .用 1 %TritonX -1 0 0抽提精子全蛋白 ,采用SDS 聚丙烯酰胺凝胶电泳结合免疫印迹实验 (Westernblot)发现单克隆抗体 6B1 0所识别的抗原为 5 0ku的蛋白 ,此蛋白为精子顶体反应发生过程中的关键蛋白之一 ,可能成为免疫灭鼠与免疫避孕的候选疫苗 相似文献
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