Yoshida M, Ishigaki K, Pursel V G
Faculty of Agriculture, Shizuoka University, Japan.
Mol Reprod Dev. 1992 Jan;31(1):68-71. doi: 10.1002/mrd.1080310112.
The present study was carried out to examine the effect of maturation media on male pronucleus formation of pig oocyte matured and fertilized in vitro. Follicular oocytes collected from prepubertal gilts at a local slaughter house were cultured (36 h) in three different media (mTCM-199, Waymouth MB 752/l, and mTLP-PVA), fertilized in vitro, and assessed for nuclear maturation and male pronucleus formation. The addition of 10% (v/v) pig follicular fluid (pFF) to maturation media significantly increased the rate of nuclear maturation of pig oocytes (P less than 0.01), whereas the rate of nuclear maturation of pig oocytes among three different media did not differ. However, the rate of male pronucleus formation of pig oocytes was significantly higher in pig oocytes matured in Waymouth MB 752/l with or without pFF than in oocytes matured in the other two media (P less than 0.01). In experiment 2, the addition of cysteine (the same concentration as in Waymouth medium, 0.57 mM), to mTLP-PVA significantly increased the rate of male pronucleus formation of pig oocytes compared with the control (P less than 0.01). The results indicate that the composition of maturation medium affects the ability of pig oocytes to form male pronuclei following sperm penetration; media containing a high concentration of cysteine (possibly as a substrate of glutathione), such as Waymouth MB 752/l, can remarkably promote this ability.
本研究旨在探讨成熟培养液对体外成熟和受精的猪卵母细胞雄原核形成的影响。从当地屠宰场的青春期前后备母猪收集卵泡卵母细胞，在三种不同的培养液（mTCM - 199、Waymouth MB 752/l和mTLP - PVA）中培养（36小时），进行体外受精，并评估核成熟和雄原核形成情况。向成熟培养液中添加10%（v/v）猪卵泡液（pFF）显著提高了猪卵母细胞核成熟率（P < 0.01），而三种不同培养液中猪卵母细胞核成熟率无差异。然而，在添加或不添加pFF的Waymouth MB 752/l中成熟的猪卵母细胞雄原核形成率显著高于在其他两种培养液中成熟的卵母细胞（P < 0.01）。在实验2中，与对照组相比，向mTLP - PVA中添加半胱氨酸（与Waymouth培养液中相同浓度，0.57 mM）显著提高了猪卵母细胞雄原核形成率（P < 0.01）。结果表明，成熟培养液的成分会影响猪卵母细胞在精子穿透后形成雄原核的能力；含有高浓度半胱氨酸（可能作为谷胱甘肽的底物）的培养液，如Waymouth MB 752/l，可显著促进这种能力。
