Schramm R D, Tennier M T, Boatman D E, Bavister B D
Wisconsin Regional Primate Research Center, University of Wisconsin, Madison 53715.
Biol Reprod. 1993 Feb;48(2):349-56. doi: 10.1095/biolreprod48.2.349.
Specific aims of this study were to compare relationships between size of intact antral follicles and meiotic competence, diameters, and chromatin configurations of germinal vesicle (GV) oocytes in non-gonadotropin-stimulated rhesus monkeys. Intact antral follicles were dissected from excised ovaries of nine normally cycling monkeys in the follicular phase of the menstrual cycle and of two acyclic monkeys. Follicles were classified according to diameter: I (200-450 microns), II (451-700 microns), III (701-1000 microns) and IV (> 1000 microns). Cumulus-enclosed oocytes were released from follicles and either measured (diameter) and fixed immediately or cultured for 48 h in modified CMRL medium containing 0.5 micrograms/ml ovine FSH, 10 micrograms/ml ovine LH, and 10% bovine calf serum. Following Hoechst staining, three distinct patterns of chromatin organization (GV1-3) were identified in GV oocytes according to the degree of association with the nucleolar periphery (encapsulation or "rimming"). In antral follicles > 450 microns in diameter, perinucleolar encapsulation (GV3) of oocytes before culture and meiotic maturation (metaphase II) of oocytes after culture increased (p < 0.01) with antral follicle growth in a graded fashion. While 56.3% of oocytes from large (> 1000 microns) follicles completed maturation, few (9.3%) oocytes from small (200-450 microns) follicles were competent to mature in vitro. At 0 h of culture, class IV follicles contained a greater (p < 0.01) proportion of GV3 oocytes and a smaller (p < 0.01) proportion of GV1 oocytes than classes I, II, and III follicles.(ABSTRACT TRUNCATED AT 250 WORDS)
本研究的具体目的是比较未用促性腺激素刺激的恒河猴中,完整窦卵泡大小与卵母细胞减数分裂能力、直径及生发泡(GV)期卵母细胞染色质构型之间的关系。从9只处于月经周期卵泡期的正常周期性恒河猴以及2只无周期恒河猴切除的卵巢中解剖出完整窦卵泡。卵泡根据直径分类:I(200 - 450微米)、II(451 - 700微米)、III(701 - 1000微米)和IV(> 1000微米)。从卵泡中释放出卵丘包裹的卵母细胞,立即测量(直径)并固定,或在含有0.5微克/毫升羊促卵泡素、10微克/毫升羊促黄体素和10%小牛血清的改良CMRL培养基中培养48小时。经Hoechst染色后,根据与核仁周边的关联程度(包绕或“边缘化”),在GV期卵母细胞中鉴定出三种不同的染色质组织模式(GV1 - 3)。在直径> 450微米的窦卵泡中,培养前卵母细胞的核仁周围包绕(GV3)以及培养后卵母细胞的减数分裂成熟(中期II)随着窦卵泡生长呈分级增加(p < 0.01)。虽然来自大卵泡(> 1000微米)的卵母细胞中有56.3%完成成熟,但来自小卵泡(200 - 450微米)的卵母细胞中很少(9.3%)有能力在体外成熟。在培养0小时时,IV类卵泡中GV3卵母细胞的比例高于I、II和III类卵泡(p < 0.01),而GV1卵母细胞的比例低于I、II和III类卵泡(p < 0.01)。(摘要截断于250字)