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与种马精子共培养对马输卵管上皮细胞从头蛋白质合成和分泌的影响。

Effect of coculture with stallion spermatozoa on de novo protein synthesis and secretion by equine oviduct epithelial cells.

作者信息

Thomas P G, Ignotz G G, Ball B A, Brinsko S P, Currie W B

机构信息

Department of Clinical Sciences, Cornell University, Ithaca, NY 14853, USA.

出版信息

Am J Vet Res. 1995 Dec;56(12):1657-62.

PMID:8599529
Abstract

Adhesion of equine spermatozoa to homologous oviduct epithelial cells (OEC) in vitro results in specific changes in spermatozoa and OEC function. To test the hypothesis that adhesion of spermatozoa affects protein synthesis and secretion by OEC, the following treatment groups were established in culture: OEC with culture medium only; control spermatozoa in culture medium only; OEC in coculture with spermatozoa; and OEC and spermatozoa in coculture, but physically separated by a microporous membrane. The experiment was replicated within each of 4 ejaculates from 3 stallions. De novo protein secretion by OEC was measured and compared by incorporation of [35S]methionine, and evaluated, using two-dimensional polyacrylamide gel electrophoresis and fluorography. Monolayers of OEC secreted a large number of proteins of molecular mass ranging from 14 to 205 kd. Adhesion of spermatozoa consistently caused reduced synthesis of 2 OEC secretory proteins and new or increased synthesis of 6 proteins. When spermatozoa and OEC were separated by a microporous membrane, some but not all of these changes were duplicated. Synthesis of 3 OEC secretory proteins, unaffected by binding of spermatozoa, was reduced when spermatozoa were prevented from contact with OEC by a microporous membrane. Adhesion of equine spermatozoa to homologous OEC monolayers and presence of equine spermatozoa resulted in qualitative and quantitative changes in synthesis and secretion of proteins by OEC. These changes have implications for storage, longevity, and maturation of spermatozoa.

摘要

体外培养时,马精子与同种输卵管上皮细胞(OEC)的黏附会导致精子和OEC功能发生特定变化。为了验证精子黏附会影响OEC蛋白质合成和分泌这一假说,在培养中设立了以下处理组:仅含培养基的OEC;仅含培养基的对照精子;与精子共培养的OEC;以及共培养但被微孔膜物理分隔的OEC和精子。该实验在3匹种马的4份精液中每份都进行了重复。通过掺入[35S]甲硫氨酸来测量和比较OEC的从头蛋白质分泌,并使用二维聚丙烯酰胺凝胶电泳和荧光自显影进行评估。OEC单层分泌大量分子量在14至205kd之间的蛋白质。精子的黏附始终导致2种OEC分泌蛋白的合成减少,以及6种蛋白质的新合成或合成增加。当精子和OEC被微孔膜分隔时,这些变化中的一些(但不是全部)会重复出现。当通过微孔膜阻止精子与OEC接触时,3种不受精子结合影响的OEC分泌蛋白的合成减少。马精子与同种OEC单层的黏附以及马精子的存在导致OEC蛋白质合成和分泌在质量和数量上发生变化。这些变化对精子的储存、寿命和成熟具有影响。

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