Johnston L A, Parrish J J, Monson R, Leibfried-Rutledge L, Susko-Parrish J L, Northey D L, Rutledge J J, Simmons L G
Department of Meat and Animal Science, University of Wisconsin, Madison 53706.
J Reprod Fertil. 1994 Jan;100(1):131-6. doi: 10.1530/jrf.0.1000131.
A study was conducted to evaluate the potential of rescuing immature oocytes from the ovaries of an endangered wild bovid, the gaur (Bos gaurus). Recovered, immature gaur oocytes (n = 59) placed in culture were evaluated for: (1) nuclear maturation after 22 h of culture, (2) fertilization with either thawed homologous (gaur) or heterologous (Bos taurus) spermatozoa 18 h after insemination and (3) embryo development. Gaur oocytes (n = 6) evaluated by fixation and staining at 22 h had all matured to metaphase II in vitro. Insemination of gaur oocytes in vitro resulted in normal fertilization (defined as the presence of spermatozoa head or two pronuclei) and embryo development to the two- and four-cell stage of 53.6% (15 of 28) and 50.0% (9 of 18), respectively, using homologous spermatozoa. The incidence of normal fertilization of in vitro matured (IVM) gaur oocytes with heterologous spermatozoa was 53.8% (7 of 13). Insemination of domestic cow oocytes in vitro resulted in normal fertilization and embryo development of 41.7% (45 of 108) and 60.0% (12 of 20), respectively, using heterologous spermatozoa. Two of four gaur embryos (50%) developed to the blastocyst stage by day 7. Embryo transfer of these two conspecific gaur blastocysts into two Holstein recipients resulted in one confirmed pregnancy. One live-born calf was delivered by Caesarean section 308 days after embryo transfer. These results demonstrate the potential of combined IVM and IVF for recovering immature germplasm from an endangered species. Specifically, immature gaur ovarian oocytes are capable of in vitro maturation and fertilization with thawed homologous spermatozoa.(ABSTRACT TRUNCATED AT 250 WORDS)
开展了一项研究,以评估从濒危野生牛科动物白肢野牛(Bos gaurus)的卵巢中挽救未成熟卵母细胞的可能性。对回收的置于培养中的未成熟白肢野牛卵母细胞(n = 59)进行了以下评估:(1)培养22小时后的核成熟情况;(2)授精18小时后用解冻的同源(白肢野牛)或异源(Bos taurus)精子进行受精;(3)胚胎发育情况。通过固定和染色在22小时评估的白肢野牛卵母细胞(n = 6)在体外均成熟至中期II。使用同源精子,体外对白肢野牛卵母细胞进行授精导致正常受精(定义为存在精子头部或两个原核),胚胎发育至二细胞期和四细胞期的比例分别为53.6%(28个中的15个)和50.0%(18个中的9个)。体外成熟(IVM)白肢野牛卵母细胞与异源精子的正常受精率为53.8%(13个中的7个)。使用异源精子,体外对家牛卵母细胞进行授精导致正常受精和胚胎发育的比例分别为41.7%(108个中的45个)和60.0%(20个中的12个)。四个白肢野牛胚胎中有两个(50%)在第7天发育至囊胚期。将这两个同种白肢野牛囊胚移植到两个荷斯坦受体中,有一个确认怀孕。胚胎移植308天后通过剖腹产分娩出一头活产犊牛。这些结果证明了体外成熟和体外受精相结合在从濒危物种中回收未成熟种质方面的潜力。具体而言,未成熟的白肢野牛卵巢卵母细胞能够进行体外成熟并与解冻的同源精子受精。(摘要截短至250字)