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延长型精子细胞对大鼠支持细胞SGP - 1、SGP - 2和CP - 2分泌及合成的调节

Regulation of the secretion and synthesis of rat Sertoli cell SGP-1, SGP-2 and CP-2 by elongate spermatids.

作者信息

McKinnell C, Sharpe R M

机构信息

MRC Reproductive Biology Unit, Edinburgh, UK.

出版信息

Int J Androl. 1997 Jun;20(3):171-9.

PMID:9354187
Abstract

The aim of this study was to investigate the effect of the absence of elongate spermatids (ES) from the rat seminiferous epithelium on the quantitative secretion and synthesis of the three major Sertoli cell secretory proteins--SGP-1, SGP-2 and CP-2. Seminiferous tubules (ST) were isolated (a) from normal 28-day-old rats, in which the most mature germ cell type is the round spermatid, (b) from normal adult rats at stages IX-XIV of the spermatogenic cycle, i.e. after spermiation, or at stages I-V and VI-VIII, when ES are still attached to the Sertoli cell, and (c) at stages VI-VIII from normal adult rats and from rats treated with methoxyacetic acid (MAA) in order to specifically deplete ES at these stages. Two-dimensional SDS PAGE combined with computerized image analysis was used to analyse 35S-methionine-labelled intracellular and secreted proteins. In the case of SGP-1 and SGP-2, almost all of the protein synthesized by ST was secreted. The total amount of both SGP-1 and CP-2 secreted by unstaged ST from immature rats was significantly lower than that secreted by unstaged ST from adult rats. The total amount of SGP-1 and CP-2 secreted by adult ST at stages IX-XIV of the spermatogenic cycle also declined dramatically compared to ST at earlier stages. The proportion of the total CP-2 synthesized by ST which was secreted also declined in all situations in which ES were absent from the seminiferous epithelium. The synthesis of only SGP-2 was changed by ES depletion from ST at stages VI-VIII, which was almost doubled compared to synthesis of this protein by ST from control rats. Our results suggest strongly that the secretion of SGP-1 and SGP-2 is via the constitutive pathway, and that regulation of these two proteins by ES is at the level of protein synthesis. In contrast, the regulation of CP-2 by ES is predominantly at the level of secretion, suggesting that this protein is secreted via a regulated pathway. Our findings add to the evidence showing that ES play a major role in the regulation of Sertoli cell function.

摘要

本研究的目的是探讨大鼠生精上皮中缺乏长形精子细胞(ES)对支持细胞三种主要分泌蛋白——SGP-1、SGP-2和CP-2的定量分泌及合成的影响。从以下几种情况分离出生精小管(ST):(a)从正常28日龄大鼠分离,其中最成熟的生殖细胞类型是圆形精子细胞;(b)从处于生精周期IX-XIV阶段的正常成年大鼠分离,即精子排出后,或从ES仍附着于支持细胞的I-V阶段和VI-VIII阶段的正常成年大鼠分离;(c)从正常成年大鼠以及用甲氧基乙酸(MAA)处理的大鼠在VI-VIII阶段分离,以便在这些阶段特异性去除ES。采用二维SDS-PAGE结合计算机图像分析技术分析35S-甲硫氨酸标记的细胞内和分泌蛋白。就SGP-1和SGP-2而言,ST合成的几乎所有蛋白都被分泌。未分阶段的未成熟大鼠ST分泌的SGP-1和CP-2总量显著低于未分阶段的成年大鼠ST分泌的量。与早期阶段的ST相比,成年ST在生精周期IX-XIV阶段分泌的SGP-1和CP-2总量也显著下降。在生精上皮中缺乏ES的所有情况下,ST合成的总CP-2中分泌的比例也下降。仅SGP-2的合成在VI-VIII阶段因ST中ES的缺失而发生变化,与对照大鼠ST合成该蛋白相比,几乎增加了一倍。我们的结果强烈表明,SGP-1和SGP-2的分泌是通过组成型途径,并且ES对这两种蛋白的调节是在蛋白质合成水平。相反,ES对CP-2的调节主要在分泌水平,表明该蛋白是通过调节型途径分泌。我们的发现进一步证明ES在支持细胞功能调节中起主要作用。

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