Expression of clusterin/sulfated glycoprotein-2 under conditions of heat stress in rat Sertoli cells and a mouse Sertoli cell line.
Expression of clusterin/sulfated glycoprotein-2 under conditions of heat stress in rat Sertoli cells and a mouse Sertoli cell line.
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DOI：
10.1002/j.1939-4640.1997.tb01917.x
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发表时间：
1997-05
影响因子：
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通讯作者：
Ann M. Clark;Michael D. Griswold
中科院分区：
文献类型：
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作者：
Ann M. Clark;Michael D. Griswold
睾丸素是大鼠支持细胞产生的主要蛋白质，并沉积在精子膜上;然而，其功能尚不清楚。为了深入了解支持细胞中丛生素的调节，本研究的目的是建立支持细胞中丛生素表达受到影响的体外模型。将大鼠支持细胞和小鼠支持细胞（MSC 1）在热应激条件（41 ℃）下培养长达48小时。将Sertoli细胞中丛生蛋白的mRNA与人上皮样癌细胞（A431）中的mRNA进行比较，以确定丛生蛋白表达是否以lestis特异性方式调节。还检查了热休克蛋白70（HSP 70）的mRNA，因为它是已知的应激调节基因。热应激开始后4小时，三种细胞中HSP 70 mRNA的表达均增加。A431细胞热应激4 h后，A431细胞中A431蛋白mRNA的表达明显高于对照组（P < 0.05），而MSC 1和Sertoli细胞中A431蛋白mRNA的表达直到12 h才明显高于对照组（P < 0.05）。MSC 1细胞中丛生蛋白mRNA的诱导持续至少48小时，并且需要细胞持续暴露于41摄氏度的温度。丛生蛋白mRNA量的增加不是由于转录物半衰期的增加，如通过向对照与热应激MSC 1细胞的培养基中添加放线菌素D所确定的。从这个体外模型的发展，我们已经看到，clusterin诱导热应激的时间是支持细胞特异性的，是不同于热休克蛋白70。在热应激期间存活细胞中的这种反应可能是保护性的，因为丛生蛋白会结合毒性化合物或溶解退化细胞释放的细胞碎片。
Clusterin is the major protein produced by rat Sertoli cells and is deposited onto sperm membranes; however, its function is unknown. In order to gain insight into the regulation of clusterin in Sertoli cells, the objective of the present study was to develop a model where the expression of clusterin could be affected in Sertoli cells in vitro. Rat Sertoli cells and mouse Sertoli cells (MSC1) were cultured under heat stress conditions (41 degrees C) for up to 48 hours. The mRNA for clusterin in Sertoli cells was compared to that in human epitheliod cancer cells (A431) to determine if clusterin expression was regulated in a lestis-specific manner. The mRNA for heat shock protein 70 (HSP70) was also examined as it is a known stress-regulated gene. Expression of HSP70 mRNA was increased in all three cell types by 4 hours after the start of heat stress. Clusterin mRNA was increased over that of controls by 4 hours in heat-stressed A431 cells but did not significantly increase in MSC1 or Sertoli cells until 12 hours (P < 0.05). The induction of clusterin mRNA in MSC1 cells continued for at least 48 hours and required the sustained exposure of cells to the 41 degrees C temperature. The increase in the amount of clusterin mRNA was not due to an increase in transcript half-life, as determined by the addition of actinomycin D to the media of control vs. heat-stressed MSC1 cells. From the development of this in vitro model, we have seen that the timing of induction of clusterin by heat stress is Sertoli cell specific and is different than that of HSP70. This response in surviving cells during heat stress may be protective in that clusterin would bind to toxic compounds or solubilize cellular debris released by degenerating cells.