Growth hormone size variants: changes in the pituitary during development of the chicken.

C. Arámburo, M. Luna, M. Carranza, M. Reyes, H. Martínez-Coria, C. Scanes
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引用次数: 21

Abstract

There is considerable evidence for the existence of structural variants of growth hormone (GH). The chicken is a useful model for investigating GH heterogeneity as both size and charge immunoreactive-(ir) variants have been observed in the pituitary and plasma. The present study examined the size distribution of ir-GH in the pituitary gland of chicken, from late embryogenesis through adulthood. Pituitaries were homogenized in the presence of protease inhibitor, and the GH size variants were separated by SDS-PAGE, transferred by Western blotting, immunostained with a specific antiserum to chicken GH, and quantitated by chemiluminescence followed by laser densitometry (chemiluminescent assay). Under nonreducing conditions ir-GH bands of 15, 22, 25, 44, 50, 66, 80, 98, 105 and >110 kDa were observed. Both the relative proportion of the GH size variants and the total pituitary content varied with developmental stage and age. The proportion of the 15-kDa fragment was greatest in the embryonic stage, and then it decreased. The proportion of the monomeric 22-kDa form was lowest at 18 days of embryogenesis (dE) and highest at 20 dE. In contrast, the high MW forms (>/=66 kDa) were lowest in embryos, and they increased (P < 0.05) after hatching. The 22-, 44-, 66-, and 80-kDa forms were assayed for activity by radioreceptor assay following isolation by semipreparative SDS-PAGE. Only the 22-kDa GH variant showed radioreceptor activity. Under reducing conditions for SDS-PAGE, ir-GH bands of 13, 15, 18, 23, 26, 36, 39, 44, 48, 59 and 72 kDa were oberved, but most of the high MW form disappeared. There was a concomitant increase in the proportion of the monomeric band and of several submonomeric forms. The present data indicate that the expression, processing, and/or release of some if not all size variants are under some differential control during growth and development of the chicken.
生长激素大小变异:鸡发育过程中脑垂体的变化。
有相当多的证据表明生长激素(GH)存在结构变异。鸡是研究生长激素异质性的有用模型,因为在垂体和血浆中观察到大小和电荷免疫反应-(ir)变异。本研究检测了从胚胎发育晚期到成年鸡脑垂体中ir-GH的大小分布。在蛋白酶抑制剂存在的情况下,垂体均质化,SDS-PAGE分离GH大小变异,Western blotting转移,特异性抗血清对鸡GH进行免疫染色,化学发光和激光密度测定(化学发光法)定量。在非还原条件下,ir-GH条带分别为15、22、25、44、50、66、80、98、105和>110 kDa。生长激素大小变异的相对比例和垂体总含量随发育阶段和年龄的变化而变化。15kda片段的比例在胚胎期最高,随后逐渐降低。22-kDa单体形态的比例在胚胎发生18 d时最低,在胚胎发生20 d时最高,而高分子量形态(>/=66 kDa)的比例在胚胎中最低,孵化后增加(P < 0.05)。在半制备SDS-PAGE分离后,用放射受体法测定22-、44-、66-和80-kDa形式的活性。只有22 kda的GH变异具有放射性受体活性。在SDS-PAGE还原条件下,可以观察到13、15、18、23、26、36、39、44、48、59和72 kDa的ir-GH谱带,但大部分高MW谱带消失。单体带和几种亚单体形式的比例也随之增加。目前的数据表明,在鸡的生长和发育过程中,一些(如果不是全部的话)大小变异的表达、处理和/或释放受到某种差异控制。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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