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犬卵母细胞体外成熟及其体外受精研究进展 总被引：1，自引：0，他引：1
犬科动物胚胎对于改进犬科珍贵物种及保护濒危犬种十分重要,因此对犬科动物体外受精(IVF)以及辅助生殖(ART)的研究逐渐变得必要.通过体外受精和核移植已经成功获得能够发育的犬科动物胚胎,克隆的犬胚胎移植到受体母犬能成功产仔,但是这一技术的效率仍然很低.主要原因是犬科动物胚胎在体外发育能力很低.犬卵母细胞早期发育和其他动物不同,犬卵巢排出的卵母细胞处于生发泡(GV)期,在输卵管中恢复减数分裂.犬卵母细胞核难以观测并且不容易确定排卵时间,所以很难确定精子入卵的确切时间,并且多精受精的情况时常发生.回顾犬卵母细胞体外成熟(IVM)和体外受精取得的进展,对于提高其胚胎生产效率很有必要. 相似文献
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Preimplantation development of embryos in labrador retrievers 总被引：1，自引：0，他引：1
Abe Y Suwa Y Yanagimoto-Ueta Y Suzuki H 《The Journal of reproduction and development》2008,54(2):135-137
Preimplantation development of canine embryos is not well understood. To understand the timing of preattachment embryogenesis relative to the luteinizing hormone (LH) surge, early embryonic development was examined in Labrador Retrievers after artificial insemination. The embryos migrated from the oviduct to the uterus beginning on day 11 after the LH surge. This transport must be completed within 24 h. By day 13 after the LH surge, all of the embryos had moved and were localized in the uterus. The embryos developed to the morula stage within 11-13 days and to the blastocyst stage within 14 days after the LH surge, respectively. These findings add to the current understanding concerning the physiology of preimplantation development and should help further develop assisted reproductive techniques in canine species, such as cryopreservation and subsequent embryo transfer. 相似文献
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C Lavilla-Apelo H Kida H Kanagawa 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》1992,54(2):335-340
Zona-intact and zona-free mouse embryos at the morula stage were exposed to Sendai virus in vitro, and then transferred to the uteri of recipient foster mothers. Both embryos developed into expanded blastocyst stage after 48 hr of culture. Zona-intact embryos were resistant to infection and subsequent transfer resulted in the development of fetuses, indicating that the zona pellucida plays a role of a barrier to virus infection. On the other hand, zona-free embryos were susceptible to infection and only one fetus out of 64 transfers developed to term. Implantation sites were scarcely observed in the uteri of the foster mothers that received zona-free embryos, suggesting that most of the embryos did not develop after embryo transfer. Sendai virus was shed in the culture fluid of the zona-free embryos indicating viral replication in the embryonic cells. By immunofluorescence assay, viral antigens were detected in the embryos, tissues of the fetus and implantation site derived from the zona-free embryos. These findings indicate that replication of Sendai virus in the embryonic cells interfere with early embryonic development and fetal growth of the embryo. 相似文献
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为研究貉早期胚胎发育体内微环境变化,试验选用年龄相同、体重相似的成年健康母貉23只,在繁殖季节自然交配,并受配1～2次,以第1次交配为零点开始计时,分别于初配后29～99h(n=7)、100～126h(n=7)、169～268h(n=9)随机处死貉,用免疫组化、透射及扫描电镜的方法研究貉早期胚胎发育过程中输卵管和子宫的形态学和超微结构变化;用X射线能谱技术对貉输卵管液和子宫液中的钾、钙、铁、锌等9种元素进行测定。结果显示:①随着貉早期胚胎的发育,其后期输卵管长度、黏膜厚度、皱襞高度和上皮高度均显著减小(P<0.05),输卵管直径有所降低,但差异不显著(P>0.05);子宫黏膜厚度、皱襞高度及子宫腺密度均显著增加(P<0.05),子宫的直径和长度有所增加,但差异不显著(P>0.05)。②随着貉早期胚胎的发育,貉子宫黏膜上皮微绒毛、脂滴和溶酶体含量增多。③随着貉早期胚胎的发育,其后期输卵管液中硫、钙、铁、铜和锌元素含量均显著升高(P<0.05),磷含量显著降低(P<0.05),而钾、氯和钠含量呈波动性变化;子宫液中硫、氯、钾的相对含量逐渐降低(P>0.05),锌的相对含量显著降低(P<0.05),磷、钙、铁和铜的含量呈波动性变化。本试验初步揭示了貉早期胚胎发育内环境的变化,为貉胚胎体外培养体系的建立提供参考。 相似文献
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The infertility of many cows could be treated by in vitro fertilisation. In the present study laparoscopy was utilised to recover the in vivo matured oocytes from the ovary of a standing donor. After the capacitation of fresh semen with high ionic strength medium and in vitro fertilisation, a rabbit oviduct was employed as an incubator for four to five days, in order to obtain sufficiently aged embryos to be transferred to the uterus of a recipient. Using surgical or non-surgical transfer six calves were obtained. 相似文献
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为探讨猪体内、外成熟卵母细胞对核移植重组胚胎发育能力的影响,试验通过激素促排获得体内成熟卵母细胞和收集废弃卵巢获取体外成熟的卵母细胞,分别构建核移植重组胚,比较其卵裂率、囊胚率及胚胎移植受孕情况。结果显示,PGC+PMSG+HCG组的平均排卵数（27.8枚/头）显著高于PGC+HCG （12.5枚/头）、PMSG+HCG （13.7枚/头）及自然发情组（11.5枚/头）（P<0.05）,体内收集到的卵母细胞,可用于构建核移植重组胚的可用卵率均达到90%以上,与其他处理组差异不显著（P>0.05）,说明通过激素处理可获得更多的可用卵母细胞,而且卵母细胞的质量没有显著差异;以体内和体外成熟卵母细胞作为核移植受体构建的克隆胚胎,二者的胚胎融合率（80.31%和79.29%）和卵裂率（90.40%和86.51%）差异均不显著（P>0.05）,但来自体内成熟卵母细胞克隆的胚胎发育至囊胚期的比例显著升高（P<0.05）;将体内、外成熟卵母细胞构建的核移植重组胚分别移植代孕母猪,头平均移植30或60枚时,体内成熟卵母构建的克隆胚胎移植出生仔猪10头,而体外培养卵母细胞构建的克隆胚胎均未着床受孕,表明通过激素促排获得的卵母细胞质量更好,能显著提高克隆胚胎的囊胚率,减少胚胎移植数量,提高代孕母猪的怀孕率。 相似文献
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Techakumph M Sukavong Y Yienvisavakul V Buntaracha B Pharee S Intaramongkol S Apimeteetumrong M Intaramongkol J 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2001,63(8):849-852
To investigate the development of fresh and frozen swamp buffalo embryos after transfer to synchronized recipients, 14 fresh embryos and 28 frozen embryos, collected from Thai swamp buffalo cows of an elite herd at the Surin Breeding Center, were transferred nonsurgically to 31 synchronized recipients buffalo cows. One fresh embryo was transferred to each of 14 recipients. Twenty eight frozen embryos were transferred to 17 recipients of which 7 cows received I embryo, 9 cows received 2 embryos and 1 cow received 3 embryos. Pregnancy was diagnosed by real time B-mode ultrasonography one month after transfer and confirmed by rectal palpation one and two months later. The pregnant cows were kept under observation until calving. The results of fresh embryo transfer showed that 5/14 (35.7%) were pregnant after 30 days, 4/14 (28.6%) remained pregnant until the 3rd month and 2/14 (14.3%) calved. With the frozen embryos, only one cow which received three embryos became pregnant and remained so for 3 months although the embryo did not survive to full term. The overall pregnancy rate using frozen embryos was 5.9% (1/17). The study demonstrated the possibility of performing embryo transfer in elite buffalo herds for genetic improvement, however the use of frozen embryos needed further investigation. 相似文献
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HUA Zai-dong GUO Shuai XIAO Hong-wei REN Hong-yan ZHANG Li-ping GE Yao-wen 《中国畜牧兽医》2017,44(11):3143-3148
To investigate the impact of porcine oocytes in vivo and in vitro maturation (IVM) on the development of porcine somatic cell cloned embryos,the somatic cell cloned embryos cultured in vitro and the sows were treated with hormones to collect mature oocytes in vivo,and the cleavage rate, blastocyst rate and embryo implantation were compared. The results showed that the average number of ovulation in PGC+PMSG+HCG group was significantly higher than that of PGC+HCG,PMSG+HCG and the natural estrus groups (P<0.05). The oocytes collected in vivo could be used for the construction, and the available oocytes rate reached more than 90%,and there was no significant difference among the four groups (P>0.05),which indicated that groups treated by hormone could obtain more available oocytes and the quality of oocytes was not significant different. In vivo and in vitro matured oocytes were used as nuclear transfer embryos of recombinant receptor,the fusion efficiency (80.31% and 79.29%) and cleavage rate (90.40% and 86.51%) were not significant different (P>0.05), but the proportion of in vivo matured oocytes cloned embryos developed into the blastocyst stage was significantly higher (P<0.05). The reconstructed embryos made from in vivo and in vitro matured oocytes were transplanted into surrogate sows (transferred 30 or 60 embryos),10 piglets were born in in vivo maturation of cloned embryo transfer group,while there was no implantation in in vitro maturation of cloned embryo transfer group. The results showed that high quality oocytes obtained by superovulation could significantly increase the blastocyst rate of embryos,reduce the number of embryos transferred and improve the pregnancy rate of surrogate sows. 相似文献
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P. Lonergan 《Acta veterinaria Scandinavica》1994,35(4):307
Development of mammalian embryos in vitro is functionally and temporally inferior to embryo development as it occurs inside the female reproductive tract. The deficiencies of cultured embryos range from slow cleavage rates to complete developmental arrests or blocks, occurring at particular stages in many species. A variety of approaches have been used to overcome the blocks, including most extensively the coculture of preimplantation embryos with various somatic cells. However, even with coculture, development of embryos in vitro is still not equivalent to that in vivo. In most laboratories, only 25–40% of inseminated oocytes develop into morulae and blastocysts in spite of numerous variations on the basic technique. A better understanding of the factors governing embryonic growth is required before we can hope to achieve results comparable with those occurring in vivo. 相似文献
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Experimental transmission of bovine viral diseases by insemination with contaminated semen or during embryo transfer 总被引：1，自引：0，他引：1
D H Schlafer J H Gillespie R H Foote S Quick N N Pennow E P Dougherty E I Schiff S E Allen P A Powers C E Hall 《DTW. Deutsche tier?rztliche Wochenschrift》1990,97(2):68-72
Three experimental approaches were used to study transmission of blue tongue (BT), infectious bovine rhinotracheitis (IBR) and bovine virus diarrhoea (BVD) viruses. These were insemination with contaminated semen, experimental infection of embryo donor cows, or transfer of embryos experimentally exposed to virus in vitro to normal recipients. Parameters assessed included number and quality of embryos produced, virus detection (isolation and electron microscopy), serology and histopathology. All superovulated sesceptible cows inseminated with semen containing blue tongue virus (BTV) (n = 2) or infectious bovine rhinotracheitis virus (IBRV) (n = 2) became infected. One cow inseminated with semen containing BTV produced seven virus-free seven-day-old embryos; the second cow failed to produce any embryos. One of two cows inseminated with semen containing IBRV produced two underdeveloped, virus-free embryos while no embryos were produced by the second cow. One of two cows inseminated with semen containing bovine viral diarrhoea virus (BVDV) became infected. Two poorly developed, virus-free seven-day-old embryos were recovered from one of these cows. Superovulated susceptible cows inoculated either intramuscularly with BTV (n = 3) or intranasally with IBR virus (n = 2) became infected. Virus was isolated from some tissues of two BTV-infected cows, neither of which produced embryos. A third BTV-infected cow produced two virus-free embryos collected at necropsy five days after inoculation. One of two cows experimentally infected with IBR virus, produced three embryos but virus was not detected either by electron microscopy (1 embryo) or in cell culture by cytopathic alterations (1 embryo).(ABSTRACT TRUNCATED AT 400 WORDS) 相似文献
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体外胚胎生产作为一项高效的辅助生殖技术,对优质种质资源的保存利用及遗传改良具有重要意义。但与体内生产胚胎相比,体外生产胚胎在培养过程中出现生长发育缓慢、卵裂率低、凋亡比例增加等问题,移植后伴随着胎盘肥大、孕期延长等问题,还可能出现早产、出生体重降低、巨胎综合征(LOS)等,这与基因表达异常和表观遗传修饰异常有重要关系。文章简单回顾了近年来表观修饰学在体内、外生产胚胎的研究进展,主要介绍了印记基因的表观修饰、DNA甲基化及组蛋白乙酰化在哺乳动物体内、外生产胚胎之间的差异,简述了微阵列技术在体内、外生产胚胎之间的应用,以期找到导致体内、外胚胎质量差异的关键印记基因及其作用途径,探讨体外生产胚胎质量低于体内胚胎的原因,进而改善体外胚胎生产体系。 相似文献
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C Feltrin CA Cooper N Mohamad‐Fauzi VHV Rodrigues LH Aguiar S Gaudencio‐Neto LT Martins CEM Calderón AS Morais IS Carneiro TM Almeida ING Silva JL Rodrigues EA Maga JD Murray AB Libório LR Bertolini M Bertolini 《Reproduction in domestic animals》2014,49(4):648-656
The presence of the zona pellucida has been perceived as a requirement for the oviductal transfer of cloned embryos at early stages of development while protecting the embryo from an immune system response. We hypothesized that steroid hormone therapy could reduce a potential cellular immune response after the transfer of zona‐free cloned embryos into the oviduct of recipient female goats. In Experiment 1, seven does were used to study the systemic immunosuppressant effect of the methylprednisolone administration (for 3 days) on blood cell counts. Whole blood was collected prior to treatment with methyprednisolone and then on Days 1, 2, 3, 4, 7, 14, 21 and 28 after the first dose of methylprednisolone for the analysis of haematological parameters. Methylprednisolone treatment significantly reduced circulating white blood cells and neutrophils in comparison with pre‐treatment levels, demonstrating a systemic immunosuppressant effect. In Experiment 2, a group of 58 does were used as recipient females to study the effect of administration of methylprednisolone for 3 days on the establishment of pregnancies after the transfer of zona‐free cloned embryos into the oviducts. No effects on pregnancy rates on Day 30 were observed regarding the distinct treatment groups (control vs. methylprednisolone), the source of oocytes (in vivo‐ vs in vitro‐matured) or the presence or absence of the zona pellucida in embryos. In summary, methylprednisolone was effective at inducing a systemic immunosuppressed state in goats, but the treatment prior to embryo transfer did not affect pregnancy rates. Moreover, pregnancy rates were similar between zona‐free and zona‐intact goat cloned embryos. 相似文献
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Production of transgenic animals and embryo cloning are only a few examples of new biotechnological methods applied to animal embryos. All these techniques require large amounts of oocytes and early embryos. In many laboratory animals, embryos matured and fertilized in vivo are easily obtained, but with larger domestic species it requires laborious surgical procedures and the number of embryos obtained remains relatively small (Bracken et al. 1982). The in vitro maturation of follicular oocytes derived from slaughterhouse ovaries and their in vitro fertilization provides large numbers of oocytes and embryos with considerably less effort. The final proof of the success in the in vitro maturation and fertilization procedure is the birth of healthy progeny. Also the normal preimplantation development of the embryos gives useful information about the efficiency of the method employed. 相似文献