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从青春期前和成年猪获取的卵母细胞中的受精和囊胚发育。

Fertilization and blastocyst development in oocytes obtained from prepubertal and adult pigs.

作者信息

Sherrer E S, Rathbun T J, Davis D L

机构信息

Department of Animal Sciences and Industry, Kansas State University, Manhattan 66506, USA.

出版信息

J Anim Sci. 2004 Jan;82(1):102-8. doi: 10.2527/2004.821102x.

Abstract

Polyspermic fertilization and embryo quality are important issues for the in vitro production of pig embryos. We hypothesized that oocyte donor (prepubertal gilt vs. sow) affects polyspermy and blastocyst development in vitro and that the sexual maturity of the oocyte donor affects the response to sperm concentration in the fertilization medium. In Exp. 1, oocytes of sows and gilts were mounted and stained 12 h after insemination to provide fertilization data. In Exp. 2, putative embryos were developed in vitro to 144 h post-insemination before mounting. In both experiments, cumulus-oocyte complexes (COC) were collected from ovaries of prepubertal gilts and adult sows. Sperm were added after maturation of COC for 40 to 44 h. Sperm from two boars at 0.5 to 5.0 x 10(6) sperm/mL was used for insemination. More (P < 0.01) monospermic fertilizations were observed in oocytes derived from gilts than for oocytes from sows. There were fewer (P < 0.02) penetrated sperm per fertilized oocyte in oocytes from gilts compared with sows. There were effects of semen donor (boar) on the percentage of monospermic (P < 0.01) and polyspermic (P < 0.002) fertilizations, and on the number of penetrated sperm/fertilized oocyte (P < 0.02). In Exp. 2, cleavage and blastocyst formation was evaluated at 2 and 6 d postinsemination, respectively. More (P < 0.001) blastocysts developed from sow-derived oocytes than from gilt-derived oocytes. More (P < 0.05) total cells per blastocyst were observed in embryos from sow-derived oocytes than from gilt-derived oocytes. Semen donor affected the percentage of oocytes cleaving (P < 0.02), and a boar x sperm concentration interaction affected (P < 0.05) the incidence of blastocyt formation. Results indicate that sexual maturity of the donor is not responsible for the high incidence of polyspermy in porcine in vitro fertilization. However, blastocyst development is improved by the use of oocytes from sows rather than from prepubertal gilts.

摘要

多精受精和胚胎质量是猪胚胎体外生产中的重要问题。我们假设卵母细胞供体(青春期前后备母猪与经产母猪)会影响体外多精受精和囊胚发育,并且卵母细胞供体的性成熟会影响其对受精培养基中精子浓度的反应。在实验1中,授精12小时后对经产母猪和后备母猪的卵母细胞进行固定和染色以获取受精数据。在实验2中,将假定胚胎在体外培养至授精后144小时再进行固定。在两个实验中,从青春期前后备母猪和成年经产母猪的卵巢中采集卵丘 - 卵母细胞复合体(COC)。COC成熟40至44小时后添加精子。使用来自两头公猪的精子,浓度为0.5至5.0×10⁶精子/毫升进行授精。与经产母猪的卵母细胞相比,后备母猪来源的卵母细胞中观察到更多(P < 0.01)的单精受精。与经产母猪相比,后备母猪来源的受精卵母细胞中每个卵母细胞穿透的精子较少(P < 0.02)。精液供体(公猪)对单精受精(P < 0.01)和多精受精(P < 0.002)的百分比以及每个受精卵母细胞穿透的精子数量有影响(P < 0.02)。在实验2中,分别在授精后2天和6天评估卵裂和囊胚形成情况。经产母猪来源的卵母细胞发育成的囊胚比后备母猪来源的更多(P < 0.001)。经产母猪来源的胚胎每个囊胚的总细胞数比后备母猪来源的更多(P < 0.05)。精液供体影响卵母细胞的卵裂百分比(P < 0.02),并且公猪×精子浓度的相互作用影响(P < 0.05)囊胚形成的发生率。结果表明,供体的性成熟并非猪体外受精中多精受精高发生率的原因。然而,使用经产母猪而非青春期前后备母猪的卵母细胞可改善囊胚发育。

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