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牛输卵管液中发情相关糖蛋白的起源

Origin of oestrus-associated glycoproteins in bovine oviductal fluid.

作者信息

Wegner C C, Killian G J

机构信息

Department of Dairy and Animal Science, Pennsylvania State University, University Park 16802.

出版信息

J Reprod Fertil. 1992 Aug;95(3):841-54. doi: 10.1530/jrf.0.0950841.

Abstract

The aim of the present study was to determine whether the synthesis of an oestrus-associated protein found in bovine oviductal fluid varies with oviductal region, stage of cycle or day of pregnancy. Explant culture was performed using oviducts recovered from naturally cycling animals either at oestrus or 12-14 days after oestrus. Three oviductal regions, the preampulla, ampulla and isthmus, were cultured individually in the presence of 20 muCi [35S]methionine in serum-free medium for 6 h at 37 degrees C. Synthesis of oestrus-associated protein was assessed by one-dimensional SDS-PAGE, fluorography and densitometry of radiolabelled bands. Significantly more oestrus-associated protein was synthesized by the ampullar region of the oviduct, although it was detected in explant culture media from both the isthmic and preampullar regions. A polyclonal antibody produced against oestrus-associated protein was used to localize the protein in paraffin-embedded sections of oviductal explant cultures and other bovine tissues. Localization of the protein in oviductal tissue sections varied with stage of cycle (oestrus > luteal > pregnant) and region of oviduct (ampulla > preampulla/isthmus). These findings indicate an effect of oviductal region and hormonal state (cycling versus pregnant) on the synthesis and secretion of the oestrus-associated protein. Lectin affinity studies indicated that galactosyl(beta 1,3)N-acetylgalactosamine and N-acetylglucosamine residues are associated with the oestrus-associated protein.

摘要

本研究的目的是确定牛输卵管液中发现的一种发情相关蛋白的合成是否随输卵管区域、周期阶段或妊娠天数而变化。使用从自然发情周期动物发情期或发情期后12 - 14天回收的输卵管进行外植体培养。将输卵管的三个区域,即壶腹前区、壶腹和峡部,分别在含有20微居里[35S]甲硫氨酸的无血清培养基中于37℃培养6小时。通过一维SDS - PAGE、放射自显影和放射性标记条带的光密度测定来评估发情相关蛋白的合成。尽管在峡部和壶腹前区的外植体培养基中都检测到了发情相关蛋白,但输卵管壶腹区域合成的发情相关蛋白明显更多。一种针对发情相关蛋白产生的多克隆抗体被用于在输卵管外植体培养物和其他牛组织的石蜡包埋切片中定位该蛋白。该蛋白在输卵管组织切片中的定位随周期阶段(发情期>黄体期>妊娠期)和输卵管区域(壶腹>壶腹前区/峡部)而变化。这些发现表明输卵管区域和激素状态(发情周期与妊娠)对发情相关蛋白的合成和分泌有影响。凝集素亲和研究表明,半乳糖基(β1,3)N - 乙酰半乳糖胺和N - 乙酰葡糖胺残基与发情相关蛋白有关。

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