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D-甘露糖的对氨基苯基衍生物对牛精子-卵母细胞融合的抑制作用。

Inhibition of bovine sperm-oocyte fusion by the p-aminophenyl derivative of D-mannose.

作者信息

Tanghe Sofie, Van Soom A, Duchateau L, De Kruif A

机构信息

Department of Reproduction, Obstetrics and Herd Health, Faculty of Veterinary Medicine, Merelbeke, Belgium.

出版信息

Mol Reprod Dev. 2004 Feb;67(2):224-32. doi: 10.1002/mrd.10387.

Abstract

Several steps in the process of mammalian fertilization are mediated by carbohydrates. This study investigated the role of the p-aminophenyl derivative of d-mannose (APMP) during bovine fertilization. Inseminating cumulus-oocyte complexes (COCs) in the presence of increasing APMP concentrations resulted in a significant dose-dependent decrease of the fertilization rate (P < 0.05). No negative effect of 50 mM APMP on total sperm motility and progressive motility was found. Subsequently, the fertilization steps at which this blocking effect could be exerted were investigated, i.e., sperm penetration of the cumulus oophorus, sperm-zona binding, acrosome reaction, sperm-oolemma binding, and/or sperm-oocyte fusion. Inseminating cumulus-enclosed and cumulus-denuded oocytes in the presence of 50 mM APMP significantly decreased the fertilization rate to a comparable minimum level (P < 0.05). There was no significant relationship between the number of spermatozoa bound to the zona pellucida and the APMP concentration, and APMP nor d-mannosylated bovine serum albumin (BSA) suppressed or stimulated sperm acrosomal status. Inseminating zona-free oocytes in the presence of 50 mM APMP did not influence sperm-oolemma binding, but significantly inhibited sperm-oocyte fusion (P < 0.05). Preincubating zona-free oocytes with 200 microg/ml Con A but not with 50 mM APMP inhibited the sperm-oocyte fusion rate to the same extent as when the gametes were simultaneously exposed to 50 mM APMP. These data indicate that the blocking effect of APMP on bovine fertilization is mainly due to an inhibition of sperm-oocyte fusion, probably by specific obstruction of the sperm receptor sites that are responsible for the fusion process.

摘要

哺乳动物受精过程中的几个步骤由碳水化合物介导。本研究调查了D-甘露糖的对氨基苯基衍生物(APMP)在牛受精过程中的作用。在APMP浓度不断增加的情况下对卵丘-卵母细胞复合体(COC)进行授精,导致受精率显著呈剂量依赖性下降(P<0.05)。未发现50 mM APMP对精子总活力和渐进性活力有负面影响。随后,研究了可能发挥这种阻断作用的受精步骤,即精子穿透卵丘、精子与透明带结合、顶体反应、精子与卵黄膜结合和/或精子与卵母细胞融合。在50 mM APMP存在下对卵丘包被和去卵丘的卵母细胞进行授精,显著降低受精率至相当的最低水平(P<0.05)。与透明带结合的精子数量与APMP浓度之间无显著关系,APMP和D-甘露糖基化牛血清白蛋白(BSA)均未抑制或刺激精子顶体状态。在50 mM APMP存在下对无透明带卵母细胞进行授精不影响精子与卵黄膜结合,但显著抑制精子与卵母细胞融合(P<0.05)。用200 μg/ml伴刀豆球蛋白A而非50 mM APMP对无透明带卵母细胞进行预孵育,对精子与卵母细胞融合率的抑制程度与配子同时暴露于50 mM APMP时相同。这些数据表明,APMP对牛受精的阻断作用主要是由于抑制精子与卵母细胞融合,可能是通过特异性阻碍负责融合过程的精子受体位点。

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