Longo F J, Woerner M, Chiba K, Hoshi M
Department of Anatomy, University of Iowa, USA.
Zygote. 1995 Aug;3(3):225-39. doi: 10.1017/s0967199400002628.
Maturation of the starfish oocyte cortex to produce an effective cortical granule reaction and fertilisation envelope is believed to develop in three phases: (1) pre-methyladenine (1-MA) stimulation; (2) post-1-MA stimulation, pregerminal vesicle breakdown; and (3) post-germinal vesicle breakdown. The present study was initiated to identify what each of these phases may encompass, specifically with respect to structures associated with the oocyte cortex, including cortical granules, microvilli and vitelline layer. 1-MA treatment brought about an orientation of cortical granules such that they became positioned perpendicular to the oocyte surface, and an approximately 4-fold decrease in microvillar length. A-23187 activation of immature oocytes treated with (10 min; pregerminal vesicle breakdown) or without 1-MA resulted in a reduction in cortical granule number of 21% and 41%, respectively (mature oocytes underwent a 96% reduction in cortical granules). Elevation of the fertilisation envelope in both cases was significantly retarded compared with activated mature oocytes. In activated mature oocytes, the vitelline layer elevated 20.0 +/- 5.4 mu m from the egg's surface, whereas in immature oocytes treated with just A-23187 or with 1-MA (10 min) and A-23187, it lifted 0.35 +/- 0.1 and 0.17 +/- 0.04 mu m, respectively. The fertilisation envelopes of activated (or fertilised) immature oocytes also differed morphologically from those of mature oocytes. In activated, immature oocytes, the fertilisation envelope was not uniform in its thickness and possessed thick and thin regions as well as fenestrations. Additionally, it lacked a complete electron-dense stratum that characterised the fertilisation envelopes of mature oocytes. The nascent perivitelline space of immature oocytes was also distinguished by the presence of numerous vesicles which appeared to be derived from microvilli. Differences in the morphology of cortices from activated (fertilised) and non-activated, immature and mature oocytes substantiate previous investigations demonstrating three phases of cortical maturation, and are consistent with physiological changes that occur during oocyte maturation, involving ionic conductance of the plasma membrane, establishment of slow and fast blocks to polyspermy and elevation of a fertilisation envelope.
(1)甲基腺嘌呤前(1-MA)刺激;(2)1-MA刺激后、生发泡破裂前;以及(3)生发泡破裂后。本研究旨在确定这些阶段各自可能包含的内容,特别是关于与卵母细胞皮层相关的结构,包括皮层颗粒、微绒毛和卵黄膜。1-MA处理使皮层颗粒定向,使其垂直于卵母细胞表面定位,并使微绒毛长度减少约4倍。用(10分钟;生发泡破裂)或不用1-MA处理的未成熟卵母细胞经A-23187激活后,皮层颗粒数量分别减少21%和41%(成熟卵母细胞的皮层颗粒减少96%)。与激活的成熟卵母细胞相比,两种情况下受精膜的升高均明显延迟。在激活的成熟卵母细胞中,卵黄膜从卵表面升高20.0±5.4μm,而在仅用A-23187处理或用1-MA(10分钟)和A-23187处理的未成熟卵母细胞中,卵黄膜分别升高0.35±0.1和0.17±0.04μm。激活的(或受精的)未成熟卵母细胞的受精膜在形态上也与成熟卵母细胞的不同。在激活的未成熟卵母细胞中,受精膜厚度不均匀,有厚区和薄区以及小孔。此外,它缺乏成熟卵母细胞受精膜特有的完整电子致密层。未成熟卵母细胞新形成的卵周隙的特征还在于存在许多似乎源自微绒毛的小泡。激活的(受精的)与未激活的、未成熟和成熟卵母细胞的皮层形态差异证实了先前关于皮层成熟三个阶段的研究,并与卵母细胞成熟过程中发生的生理变化一致,这些生理变化涉及质膜的离子传导、对多精受精的慢阻断和快阻断的建立以及受精膜的升高。