Mattioli M, Bacci M L, Galeati G, Seren E
Istituto di Fisiologia Veterinaria, Via Belmeloro 8/2, 40126, Bologna, Italy.
Theriogenology. 1991 Jul;36(1):95-105. doi: 10.1016/0093-691x(91)90438-j.
This research was designed to investigate the effects of LH and FSH (50 ng/ml) on pig oocyte maturation in vitro. The following parameters were studied: a) the degree of heterologous coupling between cumulus cells and oocytes, evaluated by measuring the 3H-uridine and 3H-choline uptake in cumulus enclosed oocytes; b) meiotic maturation; c) cytoplasmatic maturation, evaluated by analyzing the ability of the oocytes to promote male pronucleus formation after in vitro fertilization. Despite the marked cumuli expansion induced by gonadotropins, uridine uptake was not influenced by LH or FSH. By contrast, choline uptake in LH-treated oocytes was significantly higher than in FSH-treated or control oocytes (3199 cpm+/-251 vs 1686 cpm+/-142, P<0.01). Gonadotropins accelerated meiotic progression, and after 30 hours of culture the percentage of oocytes at the germinal vesicle stage was significantly lower (P<0.01) in LH-(24%, 24/102) and FSH-(20%, 18/90) treated oocytes than in control oocytes (76%, 64/84). After 44 hours of culture, the percentage of oocytes reaching the MII stage was significantly higher (P<0.01) in the presence of LH (76%, 92/120) and FSH (86%, 92/108) than in the controls (35%, 40/116). The percentage of oocytes capable of sustaining male pronucleus formation was similar in the control (48.4%, 63/132) and FSH-treated oocytes (44.3%, 51/116), while it was markedly increased (P<0.01) by the addition of LH (72.7%, 143/197). The data reported indicate that in vitro pig oocytes tend to undergo meiotic maturation even in the absence of hormones. However, in our in vitro system, LH and FSH accelerated and facilitated meiotic progression, and LH selectively improved cytoplasmic maturation which is required to promote the formation of a male pronucleus.
本研究旨在探讨促黄体生成素(LH)和促卵泡生成素(FSH,50 ng/ml)对猪卵母细胞体外成熟的影响。研究了以下参数:a)通过测量卵丘包裹卵母细胞中3H-尿苷和3H-胆碱的摄取来评估卵丘细胞与卵母细胞之间的异源偶联程度;b)减数分裂成熟;c)细胞质成熟,通过分析卵母细胞在体外受精后促进雄原核形成的能力来评估。尽管促性腺激素诱导了明显的卵丘扩展,但尿苷摄取不受LH或FSH的影响。相比之下,LH处理的卵母细胞中胆碱摄取显著高于FSH处理的或对照卵母细胞(3199 cpm±251对1686 cpm±142,P<0.01)。促性腺激素加速了减数分裂进程,培养30小时后,LH处理组(24%,24/102)和FSH处理组(20%,18/90)卵母细胞处于生发泡期的百分比显著低于对照组(76%,64/84)(P<0.01)。培养44小时后,存在LH(76%,92/120)和FSH(86%,92/108)时卵母细胞达到MII期的百分比显著高于对照组(35%,40/116)(P<0.01)。能够支持雄原核形成的卵母细胞百分比在对照组(48.4%,63/132)和FSH处理组(44.3%,51/116)中相似,而添加LH后显著增加(P<0.01)(72.7%,143/197)。所报道的数据表明,体外培养的猪卵母细胞即使在没有激素的情况下也倾向于进行减数分裂成熟。然而,在我们的体外系统中,LH和FSH加速并促进了减数分裂进程,并且LH选择性地改善了促进雄原核形成所需的细胞质成熟。