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热带环境下婆罗门牛和荷斯坦奶牛卵母细胞中热休克蛋白70.1基因的发育能力及表达

Developmental competence and expression of the Hsp 70.1 gene in oocytes obtained from Bos indicus and Bos taurus dairy cows in a tropical environment.

作者信息

Camargo L S A, Viana J H M, Ramos A A, Serapião R V, de Sa W F, Ferreira A M, Guimarães M F M, do Vale Filho V R

机构信息

Embrapa Dairy Cattle, Juiz de Fora, MG, Brazil.

出版信息

Theriogenology. 2007 Sep 1;68(4):626-32. doi: 10.1016/j.theriogenology.2007.03.029. Epub 2007 Jun 27.

Abstract

Bos indicus cows usually have better reproductive performance in tropical and subtropical regions than Bos taurus cows, presumably due to their better adaptation to tropical environments. The aim of this study was to evaluate the developmental competence and expression of the Hsp 70.1 gene in immature oocytes from B. taurus (Holstein) and B. indicus (Gyr) dairy cows raised in a tropical region. Cumulus-oocyte complexes were obtained by transvaginal ultrasound-guided follicle aspiration between spring and early autumn, and subjected to in vitro maturation and fertilization. Presumptive zygotes were co-cultured with their own cumulus cells in CR2aa media with 10% fetal calf serum; Grade 1 blastocysts were transferred to synchronized crossbred recipients. The total RNA was extracted from immature Holstein and Gyr oocytes (three pools for each breed) and relative quantification of the Hsp 70.1 transcripts was performed by real time PCR after reverse transcription. Cleavage and blastocyst rates were greater (P<0.05) for Gyr (n=390 oocytes) than Holstein (n=505) breed (66.7% versus 53.1% of cleavage and 19.6% versus 10.8% of blastocysts, respectively), but pregnancy rates were not significantly different following transfer to recipients (44.5% for 36 Gyr embryos; 60% for 10 Holstein embryos). Holstein immature oocytes had a higher level (P<0.05) of Hsp 70.1 relative expression (1.82+/-0.22; mean+/-S.E.M.) than Gyr oocytes (1.12+/-0.11). In conclusion, Gyr oocytes obtained in a tropical region were less subject to stress and more likely to develop (after IVF) than Holstein oocytes.

摘要

瘤牛在热带和亚热带地区的繁殖性能通常优于普通牛,这可能是因为它们对热带环境的适应性更强。本研究旨在评估在热带地区饲养的普通牛(荷斯坦牛)和瘤牛(吉尔牛)未成熟卵母细胞中热休克蛋白70.1(Hsp 70.1)基因的发育能力和表达情况。在春季至初秋期间,通过经阴道超声引导下的卵泡抽吸术获取卵丘-卵母细胞复合体,并进行体外成熟和受精。将推定的受精卵与其自身的卵丘细胞在含有10%胎牛血清的CR2aa培养基中共同培养;1级囊胚被移植到同期发情的杂交受体中。从未成熟的荷斯坦牛和吉尔牛卵母细胞(每个品种三个样本池)中提取总RNA,反转录后通过实时PCR对Hsp 70.1转录本进行相对定量分析。吉尔牛(n = 390个卵母细胞)的卵裂率和囊胚率高于荷斯坦牛(n = 505个)(分别为66.7%对53.1%的卵裂率和19.6%对10.8%的囊胚率,P<0.05),但移植到受体后的妊娠率无显著差异(36个吉尔牛胚胎的妊娠率为44.5%;10个荷斯坦牛胚胎的妊娠率为60%)。荷斯坦牛未成熟卵母细胞的Hsp 70.1相对表达水平(1.82±0.22;平均值±标准误)高于吉尔牛卵母细胞(1.12±0.11,P<0.05)。总之,在热带地区获取的吉尔牛卵母细胞比荷斯坦牛卵母细胞更不易受到应激影响,并且(体外受精后)更有可能发育。

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