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在体外培养的外植体中,从周期性和早期妊娠的猪输卵管组织中从头合成和释放多肽。

De novo synthesis and release of polypeptides from cyclic and early pregnant porcine oviductal tissue in explant culture.

作者信息

Buhi W C, Vallet J L, Bazer F W

机构信息

Department of Obstetrics and Gynecology, University of Florida, Gainesville 32610.

出版信息

J Exp Zool. 1989 Oct;252(1):79-88. doi: 10.1002/jez.1402520111.

Abstract

The objective of the present study was to identify and characterize in a limited manner the major de novo oviductal secretory proteins (OSP) synthesized and released by the porcine oviduct. Oviductal tissue was collected on various days of the estrous cycle (EC) and early pregnancy (EP) and cultured in a modified minimal essential medium supplemented with 100 muCi L-[3H]-leucine. Oviductal secretory activity, as measured by the rate of incorporation of 3H-leucine (dpm/mg wet tissue weight) into nondialyzable macromolecules, was greatest (P less than .01) between days 0 and 2 and reached its lowest levels on days 10 to 15. There was no difference between left and right side or pregnancy status. This increased rate of incorporation at proestrus and estrus is temporally associated with elevated levels of estrogen. Incorporation rate for ampulla was greater than for the isthmus. Analysis of oviductal culture medium by one-dimensional sodium dodecyl sulphate-polyacrylamide gel electrophoresis and fluorography revealed three protein bands of relative molecular weight (Mr) 335,000, 115,000, and 85,000, which were associated with proestrus, estrus, and metestrus and were not detectable on other days. All three proteins also incorporated 3H-glucosamine. The 115,000 Mr band was the major 3H-glucosamine-labeled protein. Two protein bands (Mr 60,000 and 20,000) were expressed with increasing progesterone during diestrus. Other de novo synthesized protein bands appear to be present throughout the EC and EP with little modulation by estrogen or progesterone. Thus, this study demonstrates that for the porcine oviduct, the increase in the incorporation rate of 3H-leucine into OSP by both whole oviduct and ampulla and de novo synthesis and secretion of three glycoproteins, Mr 335,000, 115,000, and 85,000, were associated with proestrus and estrus when events such as fertilization and early cleavage stages of embryo development occurred.

摘要

本研究的目的是以有限的方式鉴定和表征由猪输卵管合成并释放的主要新生输卵管分泌蛋白(OSP)。在发情周期(EC)和早期妊娠(EP)的不同天数收集输卵管组织,并在补充有100μCi L-[3H]-亮氨酸的改良基本培养基中培养。通过将3H-亮氨酸(dpm/毫克湿组织重量)掺入不可透析大分子的速率来衡量的输卵管分泌活性在第0天至第2天之间最高(P小于0.01),并在第拾天至第15天达到最低水平。左右两侧或妊娠状态之间没有差异。发情前期和发情期掺入率的增加在时间上与雌激素水平的升高相关。壶腹部的掺入率高于峡部。通过一维十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和荧光自显影分析输卵管培养基,发现相对分子量(Mr)为335,000、115,000和85,000的三条蛋白带,它们与发情前期、发情期和发情后期相关,在其他日子无法检测到。所有三种蛋白质也掺入了3H-葡糖胺。115,000 Mr带是主要的3H-葡糖胺标记蛋白。在发情间期,随着孕酮增加,出现两条蛋白带(Mr 60,000和20,000)。其他新生合成的蛋白带似乎在整个EC和EP期间都存在,受雌激素或孕酮的调节很小。因此,本研究表明,对于猪输卵管,整个输卵管和壶腹部将3H-亮氨酸掺入OSP的速率增加以及三种糖蛋白Mr 335,000、115,000和85,000的新生合成和分泌与受精和胚胎发育早期分裂阶段等事件发生的发情前期和发情期相关。

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