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[精子质膜在卵母细胞受精中的作用。细胞电泳和免疫荧光显微镜研究]

[Function of the spermatozoal plasma membrane in oocyte fertilization. Cell electrophoresis and immunofluorescence microscopy studies].

作者信息

Glander H J, Herrmann K, Herold W

机构信息

Klinik und Poliklinik für Hautkrankheiten, Bereichs Medizin der Karl-Marx-Universität Leipzig.

出版信息

Zentralbl Gynakol. 1989;111(20):1353-60.

PMID:2686294
Abstract

The influence of the capacitation on the electric charge of the spermatozoal surface and the unspecific spermatozoon-oocyte-binding are not fully understood up to now. Therefore, human semen samples were divided into two subgroups of spermatozoa by the "swim up" method. Both of the subgroups were examined by means of the micro-electrophoresis and the immunofluorescence technique using antihuman fibronectin antibodies. Cellular electrophoresis: The 2 subgroups of spermatozoa prepared of one semen sample showed a significantly different electrophoretic mobility (EPM) after washing and resuspension in an isotonic glycin-HCl-buffer (pH 2.0). In contrast, after resuspension in Baker's-buffer (pH 7.4) no difference between the 2 subgroups of one semen sample was detectable. The incubation in a medium with Ca++ caused secondly alterations of the EPM with the result of a smaller difference between the two subgroups of spermatozoa. Albumin also modified the EPM of spermatozoa. Immunofluorescence technique: 69 per cent of spermatozoa with a normal shape and only 17 per cent of them exhibiting a pathological shape showed a positive fibronectin-fluorescent-band on the equatorial segment called equatorial fibronectin band (EFB). Spermatozoa separated by "swim up" from semen samples with various semen parameters showed different percentages of spermatozoa with EFB. There was a significant difference between the spermatozoa of the two subgroups after "swim up" concerning EFB.

摘要

获能对精子表面电荷及非特异性精子-卵母细胞结合的影响至今尚未完全明确。因此,采用“上游”法将人类精液样本分为两个精子亚组。利用抗人纤连蛋白抗体,通过微电泳和免疫荧光技术对这两个亚组进行检测。细胞电泳:由一份精液样本制备的两个精子亚组,在等渗甘氨酸-盐酸缓冲液(pH 2.0)中洗涤并重悬后,显示出显著不同的电泳迁移率(EPM)。相反,在贝克缓冲液(pH 7.4)中重悬后,一份精液样本的两个亚组之间未检测到差异。在含钙离子的培养基中孵育,其次会导致EPM发生变化,结果是两个精子亚组之间的差异变小。白蛋白也会改变精子的EPM。免疫荧光技术:69%形态正常的精子以及仅17%形态异常的精子在赤道段呈现出阳性纤连蛋白荧光带,称为赤道纤连蛋白带(EFB)。从具有不同精液参数的精液样本中通过“上游”法分离出的精子显示出具有EFB的精子百分比不同。“上游”后两个亚组的精子在EFB方面存在显著差异。

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