Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro
Transmission electron microscopy studies of the zona reaction in pig oocytes fertilized in vivo and in vitro
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DOI：
10.1530/rep.0.1220443
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发表时间：
2001-09-01
期刊：
影响因子：
3.8
通讯作者：
Rodriguez-Martinez, H
中科院分区：
文献类型：
--
作者：
Funahashi, H;Ekwall, H;Rodriguez-Martinez, H
本研究的目的是确定透明带的横截面透明带的猪卵母细胞体外成熟和排卵前或后精子穿在体外和体内，分别超微结构。将体外和体内（排卵）卵母细胞和受精卵（体外和体内受精）直接或经钌红和皂苷预处理后用戊二醛固定，处理，然后用透射电子显微镜检查。在用戊二醛固定的最大直径的标本的切片上测量的透明膜厚度在体内（9.19 +/-0.47 μ m）和体外（5.95 +/-0.51 μ m）卵母细胞之间存在差异。体内卵母细胞具有较厚的外部网状结构，而体外卵母细胞的外部网状结构要薄得多。这种网状外缘在体内和体外受精卵中都不太明显。由固定的透明带形成的网格的密度的明显差异是可见的外和内（ad-oolemmal）带之间。外部区域总是形成同心排列的纤维状网络，而内部区域显示出更紧凑的小梁状网格。然而，这两个区域，特别是外部网络，在反应后更加紧凑。在体外和体内受精卵之间，也观察到内卵区纤维聚集程度的明显差异，后者要高得多。这种纤维状网络在用钌红和皂苷预处理的受精卵中更清晰可见;体外受精卵具有纤维状的、径向取向的平行原纤维集，而在体内受精卵中它更聚集和小梁状。这些结果表明，猪卵母细胞在体内受精和体外受精时，卵透明膜的精细结构和精子穿透时的精子反应是不同的。此外，猪的外透明带和内透明带的超微结构具有不同的网络结构。
The aim of this study was to determine the ultrastructure of cross-sectioned zonae pellucidae of in vitro-matured and ovulated pig oocytes before or after sperm penetration in vitro and in vivo, respectively. The in vitro and in vivo (ovulated) oocytes and zygotes (fertilized in vitro and in vivo) were fixed with glutaraldehyde either directly or after pretreatment with ruthenium red and saponin, processed and then examined using transmission electron microscopy. The thickness of the zona pellucida, as measured on the section of the specimens with largest diameter fixed with glutaraldehyde, differed between the in vivo (9.19 +/- 0.47 mum) and in vitro (5.95 +/- 0.51 mum) oocytes. The in vivo oocytes had a rather thick external mesh-like structure, whereas it was much thinner in the in vitro oocytes. This mesh-like external rim was less apparent in both in vivo and in vitro zygotes. Obvious differences in the density of the lattice formed by the fixed zonae pellucidae were visible between the outer and inner (ad-oolemmal) zonae. The outer area always formed a concentrically arrayed fibrillar network, whereas the inner area showed a much more compact, trabecule-like mesh. However, both areas, but particularly the outer network, were much more compacted after the zona reaction. Clear differences in the degree of fibrillar aggregation of the inner zona area were also observed between in vitro and in vivo zygotes, being much higher in the latter. This fibrillar network was more clearly visible in the zygotes pretreated with ruthenium red and saponin; the in vitro zygotes had a fibrillar, radially oriented set of parallel fibrils, whereas it was much more aggregated and trabecule-like in the in vivo zygotes. These results demonstrate that the fine structure of the zona pellucida and the zona reaction at sperm penetration differ between pig oocytes fertilized in vivo and in vitro. Moreover, the ultrastructure of the outer and inner pig zonae pellucidae has a different network organization.