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研究旨在探讨猪卵母细胞核因子在重编程过程中发挥的作用。将体细胞引入未去核的MⅡ期卵母细胞中,构建体细胞核与卵母细胞核共存的核移植四倍体胚胎。通过分析核移植四倍体胚胎的早期发育情况探讨卵母细胞核因子对核移植四倍体胚胎早期发育的影响。结果显示,核移植四倍体胚胎、孤雌二倍体胚胎及孤雌单倍体胚胎这3组胚胎的卵裂率极显著高于核移植二倍体胚胎（P<0.01）,且核移植四倍体囊胚率及总细胞数也极显著高于核移植二倍体囊胚（P<0.01）。与通过标准核移植程序构建的核移植二倍体胚胎相比,核移植四倍体胚胎具有更强的发育能力。本研究建立了一个体细胞核与完整卵母细胞核因子物质共存的四倍体胚胎模型,有助于研究供体核与卵母细胞核之间的联系,为研究核因子在重编程过程中发挥的作用提供了平台。 相似文献
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将发育不同时期的兔胚和移核胚的卵裂球细胞核与去核的成熟卵母细胞共同组成移核胚,通过中间受体培养和移植实验检验胚胎的发育能力。结果表明,（１）兔囊胚之前各个时期的胚胎细胞核均可使移核胚发育到囊胚;（２）胚胎极化前后的卵裂球参与组成的移核胚发育到囊胚的比例无显著差异。但极化后 ６４细胞胚的卵裂球与去核卵母细胞的融合率低于极化前的 ８细胞胚胎卵裂球;（３）兔 １６细胞胚与去核的卵母细胞组成的移核胚可以发育到期,产仔率为 ３．１６％ ;（４）兔移核胚卵裂球用于连续核移植,其后 ２ 代均可发育成囊胚,其中第 １ 代移核胚与第 ２ 代移核胚发育率相似,但显著高于第 ３ 代移核胚;（５）兔移核胚和各代连续移核胚卵裂球与去核卵的融合率无显著差异。 相似文献
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水牛体细胞核移植方法比较及激活前的时间间隔对全细胞胞质内注射法核移植效果的影响 总被引：1，自引：0，他引：1
本研究旨在比较水牛2种体细胞核移植(Somatic Cell Nuclear Transfer,SCNT)方法的效果以及激活前的时间间隔对全细胞胞质内注射法(Whole-Cell Intracytoplasmic Microinjection,WCICSI)核移植效果的影响.采用水牛胎儿成纤维细胞作为供核,比较了透明带下注核法(Perivitelline Microinjection,PM)和WCICSI核移植效果.另外,试验了不同类型的供体细胞进行全细胞胞质内注射后与激活前的受体胞质的最适宜作用时间.结果,WCICSI构建核移植重构胚的成功率显著高于PM(87.1%vs 81.1%,P＜0.05),虽然其重构胚的分裂率极显著低于PM(49.5%vs 71.8%,P＜0.01),但囊胚率、核移植的效率无显著差异(P＞0.05).卵丘细胞和胎儿成纤维细胞在注射后3 h激活,重构胚的囊胚发育率最高;颗粒细胞注射后与激活前的最佳时间间隔可在1.5～3 h,但3 h是最佳的作用时间.结果表明,(1)WCICSI可用于水牛体细胞核移植的研究;(2)水牛胞质内注射供体细胞后3 h进行激活,核移植重构胚的发育效果最好. 相似文献
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核移植就是将共体核移入受体卵胞质获得重组胚的过程，核移植可以产生遗传组成相同的若干后代，前景迷人。猪胚胎核移植目前以早期卵裂球做供体，次级卵母细胞做受体。猪胚母体－合子转变（ＭＺＴ）发生于４细胞期，ＭＺＴ不影响重组胚的发育能力。核质融合及激活老头儿采电激法，可有效地使重级胚发育，卵母细胞质可使融入的核发生再规范产重排发育程序，供体细胞周期对核移胚的发育有重要影响。由于猪胚的特殊性，核移植的难度较大 相似文献
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采用端粒酶重复序列扩增(telomeric repeat amplification protocol,TRAP)-银染法,对小鼠卵母细胞、体内受精胚胎及重构胚的各阶段端粒酶活性进行测定,对PCR产物的电泳条带进行灰度值分析,计算端粒酶总产品总量。结果表明:(1)体内受精胚胎及核移植重构胚各阶段的端粒酶活性随胚胎分裂呈上升趋势,囊胚期最高(P0.05);(2)体内受精胚胎的端粒酶活性高于体细胞核移植重构胚端粒酶活性。同时进行囊胚计数并测定单卵裂球相对端粒酶活性,结果表明:单卵裂球的端粒酶活性呈下降趋势,囊胚期端粒酶活性最低(P0.05)。以上结果说明小鼠卵母细胞及体细胞核移植重构胚端粒酶活性变化与细胞发育水平及细胞发育全能性有关。 相似文献
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供体细胞培养处理方法对水牛核移植效果的影响 总被引：4，自引：1，他引：4
以经常规培养法 (DMEM 10 % FCS)、血清饥饿法 (DMEM 0 .5 % FCS培养 5～ 10 d)和 Apidicolin- APD结合血清饥饿法 (0 .1mg/ L APD培养 2 4 h,DMEM 0 .5 % FCS培养 1～ 18d)培养处理的水牛卵巢颗粒细胞和水牛成体耳部成纤维细胞作供核 ,分别采取带下注核法和胞质内注核法进行核移植。同一供核细胞各处理组间的核移植胚融合率 (以颗粒细胞作供核 )以及重组胚的囊胚发育率无明显差异 (P>0 .0 5 ) ,但经 APD 0 .5 % FCS培养处理供体细胞核移植后的分裂率显著高于其他组 (P<0 .0 5 )。用 7%乙醇处理的成体耳部成纤维细胞进行核移植 ,其重组胚的分裂率和囊胚发育率与对照组 (不含乙醇 )均无明显差异 (P>0 .0 5 )。结果表明 ,(1)血清饥饿处理水牛供体细胞对其核移植效果没有影响 ;(2 ) DNA合成抑制剂 APD结合血清饥饿培养处理水牛颗粒细胞和成体耳部成纤维细胞 ,可提高其核移植效果 ;(3)乙醇预激活处理水牛成体耳部成纤维细胞 ,对其核移植效果没有影响 相似文献
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不同核移植方法对牛体细胞核移植效率的影响 总被引：8，自引：0，他引：8
以牛皮肤成纤维细胞为供体细胞,比较了电融合法和细胞质内注射法2种核移植方法对体细胞核移植效率的影响.电融合法构建重组胚的效率显著低于细胞质内注射法(47.1%比89.0%,P＜0.01);重组胚培养36 h后的裂卵率和培养8 d时囊胚发育率无明显差异(76.4%比73.2%,11.2%比12.3%;P＞0.05),但相对于操作的卵母细胞总数而言,电融合法得到的总囊胚发育率显著低于细胞质内注射法(5.6%比10.9%,P＜0.01).结果表明,用细胞质内注射法进行体细胞核移植效率更高.将发育到桑椹胚和囊胚期的95枚核移植胚胎移植到33头受体牛,其中31头受体牛在移植后第2个情期前返情;1头受体牛在移植后75 d返情,但未进行直肠检查,无法确定妊娠情况;1头受体牛在移植后60 d未见返情,直肠检查确证妊娠,93 d受体牛流产.胚胎移植结果表明,用细胞质内注射法构建的体细胞核移植胚胎至少可以维持早期妊娠. 相似文献
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５．１．２种间核移植 种间核移植是将来源于不同种的供体细胞重组在一起进行核质杂交技术。ＭｃＧｒａｔｈ和Ｓｏｌｔｅｒ（１９８６）首先进行了不同种间小鼠受精卵原核互换试验。Ｗｏｌｆ和Ｋｒａｃｅｍｅｒ（１９９２）获得了不同属间（野牛×牛、羊×牛）的核质杂交囊胚,梅祺和邹贤刚（１９９３）以小鼠早期胚胎为供体细胞核,兔去核卵母细胞为受体胞质,获得不同目间核质杂交的囊胚,可见,如果种间核移植进一步完善,就可挽救像大熊猫这样的珍贵物种。 ５．１．３胚胎干细胞核移植 胚胎干（ＥＳ）细胞,是早期胚胎或原始生殖细胞（… 相似文献
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Intapan PM Thanchomnang T Lulitanond V Phongsaskulchoti P Maleewong W 《Veterinary parasitology》2008,157(1-2):65-71
A real-time fluorescence resonance energy transfer (FRET) PCR combined with a melting curve analysis was developed for the detection of Opisthorchis viverrini in its fish intermediate host, cyprinoid fishes. Real-time FRET PCR is based on a fluorescence melting curve analysis of a hybrid between an amplicon generated from a family of repeated DNA elements, the pOV-A6 specific probe sequence (Genbank Accession No. S80278), a 162bp repeated sequence specific to O. viverrini, and specific fluorophore-labeled probes. The real-time FRET PCR could detect as little as a single metacercaria artificially inoculated in 30 fish samples. The O. viverrini infected fishes were distinguished from non-infected fishes and from the genomic DNA of other parasites by their melting temperature. Sensitivity and specificity of this method were both 100% in the laboratory setting and it outperformed the microscopic method on field-collected samples as well. Melting curve analysis is a rapid, accurate, and sensitive alternative for the specific detection of O. viverrini infected fishes. It allows a high throughput and can be performed on small samples. The assay has not only great potential for epidemiological surveys of fish intermediate hosts but it could also be adapted as screening tool for a range of foodborne parasites in freshwater fishes. 相似文献
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G Yoshizaki T Okutsu T Morita M Terasawa R Yazawa Y Takeuchi 《Reproduction in domestic animals = Zuchthygiene》2012,47(Z4):187-192
We have revealed several unique characteristics of germ cell development using rainbow trout, including the fact that spermatogonia transplanted into the peritoneal cavity of newly hatched embryos migrate toward recipient gonads, that spermatogonia transplanted into female recipients start oogenesis and produce functional eggs and that diploid germ cells transplanted into triploid trout can complete gametogenesis. By combining these unique features of fish germ cells, we established allogeneic and xenogeneic transplantation systems for spermatogonia in several fish species. Spermatogonia isolated from the mature testes of vasa-green fluorescent protein (Gfp) transgenic rainbow trout were transplanted into the peritoneal cavity of triploid masu salmon newly hatched embryos. These spermatogonia migrated toward recipient salmon genital ridges with extending pseudopodia and were subsequently incorporated into them. We further confirmed that the donor-derived spermatogonia resumed gametogenesis and produced sperm and eggs in male and female salmon recipients, respectively. By inseminating the resulting eggs and sperm, we obtained only rainbow trout offspring in the F1 generation, suggesting that the triploid salmon recipients produced functional gametes derived only from donor trout. We further confirmed that this intra-peritoneal transplantation of germ cells is applicable to several marine fishes, which could be of benefit in the production of bluefin tuna that has a large broodstock (>100 kg) and is difficult to maintain in captivity. Gamete production of bluefin tuna could be more easily achieved by generating a surrogate species, such as mackerel, that can produce tuna gametes. 相似文献
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草鱼与青鱼、鳙鱼与团头鲂之间的细胞核移植 总被引：1，自引：0，他引：1
在草鱼与青鱼、鳊鱼与团头鲂之间进行核移植试验,获得了不同属鱼类间的草鱼细胞核和青鱼细胞质配合的核质杂种鱼——草青移核鱼幼鱼,以及不同亚科间的鳙鱼细胞核与团头鲂细胞质配合的核质杂种鱼——鳙团移核鱼,得率分别为0.9％和1.8％。核质杂种鱼的胚胎发育特征大多与受核体鱼相似,而形态特征大多与供核体鱼相似。表明胚胎发育明显受受核体细胞质的影响,而形态性状特征主要受供核体细胞核的影响。 相似文献
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[目的]建立德系乳肉兼用型西门塔尔牛群体,探索文山牛快速扩繁和生产性能提升技术。[方法]在马关县从600头文山黄牛、西本杂、安本杂和短本杂牛母牛中选择270头,用CIDR+PG法进行同期发情处理,选择黄体合格的受体,采用非手术法移植德系乳肉兼用型西门塔尔牛胚胎,在移植后60~90 d通过直肠检查法进行妊娠诊断,确定妊娠率,跟踪调查产犊情况,并测定胚胎移植所产犊牛的初生体重和主要体尺指标。[结果]受体牛同期发情处理270头,胚胎移植87头,妊娠28头,移植妊娠率32.18%;妊娠母牛中产犊17头,产犊率60.71%,产犊19头,成活16头,产犊成活率84.21%。公母犊牛平均的初生重33.00 kg,体高69.50 cm,体斜长62.19 cm,胸围72.69 cm,管围13.50 cm。[结论]首次在文山州开展了牛胚胎移植获得成功,杂交牛受体的移植妊娠率极显著或显著高于文山牛受体,秋季移植的妊娠率极显著高于春季和冬季的。提示以杂交牛为受体在秋季进行胚胎移植可有效提高移植成功率。 相似文献
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M Francisco‐Simão J Cardona‐Costa M Perez Camps F García‐Ximénez 《Reproduction in domestic animals》2010,45(6):1098-1103
Germ‐line chimaerism is a powerful technique that has proved to be useful to produce viable gametes when transplanted blastomeres colonize the germinal ridges in recipient embryos and obtaining offspring from such transplanted cells. In fish, ionizing radiations were commonly used for embryo penalization to cancelling the cell participation of recipient embryos in development and in gamete production. The ultraviolet (UV) radiation when compared with other radiation types is cheaper, easier and no special installations are required for its use. So, the aim of this work was to establish the optimal UV radiation dose to be applied in zebrafish embryos at mid‐blastula transition stage of development, in order to use them as penalized recipient embryos in futures chimaerism assays. A UV germicide lamp was used as radiation source (0.529 mW/cm2). Four exposure levels and three exposure times of UV radiation were tested. The survival rates obtained with the non‐dechorionated embryos without lid group suggested that it could be the optimal exposure level to achieve the objective proposed. With the obtained results, we concluded that this UV radiation dose for 60 and 30 s are optimal parameters to penalize recipient wild and gold strain zebrafish embryos, respectively in chimaerism assays, but without involving their survival and apparently normal development. 相似文献
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试验通过使用人工授精后绵羊作为受体进行胚胎移植,以寻找一种更加有效的方法提高绵羊胚胎移植的经济效益。试验中使用FSH对10只无角道赛特绵羊进行超数排卵处理,同时对60只受体小尾寒羊进行同期发情。供体羊在发情配种后4.5～5.0d从子宫角收集胚胎。同时,将胚胎移植到同期发情并进行人工授精的受体羊子宫内。总共有57枚可用胚移植给44只受体小尾寒羊,32只怀孕到分娩,共产下羔羊51只(无角道赛特羔28只,道赛特与小尾寒羊杂种羔23只)。此外,经人工授精但未进行手术移植的7只小尾寒羊产下15只杂种羔羊。移胚植受体妊娠率72.7%(32/44),移胚受体繁殖率118%(51/44),受体利用率88.3%(53/60)。移胚受体总妊娠率和受体利用率均显著高于常规ET组(P<0.01)。与常规胚胎移植相比,受体羊人工授精后移植胚胎不仅提高了无角道赛特母羊的繁殖率,而且提高了受体羊的利用率。 相似文献
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Joseph M Groff 《Veterinary Clinics of North America: Exotic Animal Practice》2004,7(3):705-56, vii
Similar to higher vertebrates, neoplasia is not an uncommon disease in fishes, which are the largest group of vertebrates. However,neoplasia in fishes is generally a benign condition with relatively few exceptions of malignant disease. The objective of this discussion is to provide an overview of neoplasia and the various neoplastic disease conditions in fishes according to organ system,including the few neoplasms of species that are familiar to the aquatic animal or exotic animal practitioner. The discussion also considers the various nonneoplastic lesions in fishes that may be confused with neoplasms, and treatment of neoplastic disease in fishes that is generally restricted to surgical intervention. 相似文献
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中国淡水鱼类种质资源的保护和利用 总被引：3，自引：0，他引：3
本文简要介绍了目前世界鱼类资源概况、我国鱼类资源现状，以及鱼类生物多样性的极丰富程度与易被破坏性，还介绍了造成鱼类种类减少或区系组成变化的主要原因以及造成鱼类遗传多样性减少的内在原因和外部原因；比较详细的提出了鱼类种质资源保护的目的、策略及措施。并就我国鲤科鱼类繁育群体的大小、鱼类种质的鉴定与评价及利用前景作了分析。 相似文献
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不同蔗糖浓度、供体细胞保存温度和融合温度对小鼠体细胞克隆胚胎发育的影响 总被引：1，自引：0，他引：1
以昆明小鼠为实验动物,经过超数排卵后得到卵丘卵母细胞复合体(COCs),以卵丘细胞为核供体细胞,卵母细胞为核受体细胞进行核移植,分析比较了不同蔗糖浓度对小鼠卵母细胞去核时间、去核率及重构胚体外发育的影响,并探讨了供体细胞的保存温度和融合温度对重构胚体外发育的影响.结果表明,各蔗糖浓度组的去核率均达到100%,但3%、4... 相似文献
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为研究抗菌肽添加量对杂交鲟血清和组织免疫相关指标和抗损伤能力的影响。挑选体格健壮、大小一致的杂交鲟（俄罗斯鲟Acipenser gueldenstaedtii♀×史氏鲟Acipenser schrencki Brandt♂）200尾，随机分成4组。对照组投喂基础饲料，Ⅰ、Ⅱ、Ⅲ组分别在基础饲料中添加400、800和1 200 mg/kg的抗菌肽，在（16.0±1.0）℃水温条件下养殖30 d。结果表明，摄食添加抗菌肽的杂交鲟，其血清和组织中白蛋白（ALB）和总蛋白（TP）含量随抗菌肽质量浓度的增加先增后减，其中Ⅱ组杂交鲟血清和组织中ALB和TP含量显著高于对照组（P<0.05）。血清中谷草转氨酶（GOT）和乳酸脱氢酶（LDH）活性随抗菌肽质量浓度的增加先降低后增加；Ⅰ组杂交鲟血清和组织中GOT活性显著低于其他组（P<0.05）；而Ⅲ组GOT和LDH活性显著高于其他组（P<0.05）。杂交鲟组织中总胆固醇（TCHO）含量随抗菌肽质量浓度的增加逐渐减少，Ⅲ组TCHO含量在组织中显著低于其他组（P<0.05）。溶菌酶（LZM）活性随抗菌肽质量浓度的增加而逐渐增加，Ⅱ、Ⅲ组杂交鲟心脏、脾脏和小肠中LZM活性显著高于其他组（P<0.05）。因此，在基础饲料中添加抗菌肽可有效提高杂交鲟的部分非特异性免疫能力和抗损伤能力，抗菌肽添加剂量在400~800 mg/kg时效果最佳。 相似文献