Nogueira Daniela, Albano Carola, Adriaenssens Tom, Cortvrindt Rita, Bourgain Claire, Devroey Paul, Smitz Johan
Follicle Biology Laboratory, Dutch-Speaking Free University of Brussels (VUB), Brussels, Belgium.
Biol Reprod. 2003 Sep;69(3):1042-52. doi: 10.1095/biolreprod.103.015982. Epub 2003 May 28.
This study addresses the role of cAMP hydrolytic isoenzyme phosphodiesterase type 3 (PDE 3) modulation on human oocyte maturation in vitro. Presence of phosphodiesterase type 3 A (PDE 3A) mRNA was confirmed in human germinal vesicle-stage (GV) oocytes. Making use of a selective PDE 3 inhibitor, Org 9935 (10 microM), oocytes retrieved from immature follicles were arrested in prophase I with a high efficiency for up to 72 h. Cumulus oocyte complexes (COCs) were retrieved in the follicular phase of the cycle before or after exposure to endogenous LH or hCG administration in vivo and randomly distributed into maturation medium with or without the PDE 3 inhibitor. Previous exposure of small follicles to LH activity in vivo had no influence on the arresting capacity of the PDE 3 inhibitor. Reversal from pharmacological arrest leads to a progression through meiosis in a normal time frame with formation of a well-aligned metaphase plate. Ultrastructure analysis of COC derived from follicles between 8 and 12 mm showed that the induced extension of prophase I arrest in vitro resulted in cytoplasm changes but not in apparent nuclear changes during culture.
本研究探讨了环磷酸腺苷（cAMP）水解同工酶3型磷酸二酯酶（PDE 3）调节对人卵母细胞体外成熟的作用。在人初级卵母细胞（GV）期卵母细胞中证实存在磷酸二酯酶3 A型（PDE 3A）信使核糖核酸（mRNA）。利用选择性PDE 3抑制剂奥格9935（10微摩尔），从未成熟卵泡中获取的卵母细胞可高效地停滞在减数分裂前期I长达72小时。在体内暴露于内源性促黄体生成素（LH）或人绒毛膜促性腺激素（hCG）给药之前或之后的月经周期卵泡期获取卵丘卵母细胞复合体（COC），并随机分配到含有或不含有PDE 3抑制剂的成熟培养基中。小卵泡先前在体内暴露于LH活性对PDE 3抑制剂的停滞能力没有影响。从药物停滞状态逆转导致减数分裂在正常时间范围内进行，并形成排列良好的中期板。对直径在8至12毫米之间的卵泡来源的COC进行超微结构分析表明，体外诱导的减数分裂前期I停滞延长导致培养过程中细胞质发生变化，但未导致明显的细胞核变化。
