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首页> 外文期刊>Journal of neuroendocrinology >Stress induces parallel changes in corticotrophin-releasing hormone (CRH) Transcription and nuclear translocation of transducer of regulated cAMP response element-binding activity 2 in hypothalamic CRH neurones.
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Stress induces parallel changes in corticotrophin-releasing hormone (CRH) Transcription and nuclear translocation of transducer of regulated cAMP response element-binding activity 2 in hypothalamic CRH neurones.

机译：应激在下丘脑CRH神经元中诱导促肾上腺皮质激素释放激素（CRH）转录和调节性cAMP反应元件结合活性2的换能器的核易位平行变化。

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摘要

Recent studies in vitro have shown that the cAMP response element-binding (CREB) co-activator, transducer of regulated CREB activity (TORC), is required for transcriptional activation of the corticotrophin-releasing hormone (CRH) gene. To determine the physiological importance of TORC2 regulating CRH transcription during stress, we examined the localisation of TORC2 in CRH neurones, as well as the relationship between changes in CRH heterogeneous nuclear (hn)RNA, nuclear translocation of TORC2 and binding of TORC2 to the CRH promoter. Immunohistochemistry revealed TORC2 immunoreactivity (irTORC2) in the dorsolateral (magnocellular) and dorsomedial (parvocellular) regions of the hypothalamic paraventricular nucleus (PVN). Although staining was mostly cytosolic under basal conditions, there was a marked increase in nuclear irTORC2 in the dorsomedial region after 30 min of restraint, concomitant with increases in CRH hnRNA levels. Levels of nuclear irTORC2 and CRH hnRNA had returned to basal 4 h after stress. Double-staining immunohistochemistry showed TORC2 co-staining in 100% of detected CRH neurones, and nuclear translocation after 30 min of restraint in 61%. Cellular distribution of TORC2 in the dorsolateral PVN was unaffected by restraint. Chromatin immunoprecipitation experiments showed recruitment of TORC2 and phosphorylated CREB (pCREB) by the CRH promoter after 30 min of restraint, but 4 h after stress only pCREB was associated with the CRH promoter. The demonstration that TORC2 translocates to the nucleus of hypothalamic CRH neurones and interacts with the CRH promoter in conjunction with the activation of CRH transcription during restraint stress, provides strong evidence for the involvement of TORC2 in the physiological regulation of CRH transcription.
机译：近期的体外研究表明，促肾上腺皮质激素释放激素（CRH）基因的转录激活需要cAMP反应元件结合（CREB）共激活子，即受调控的CREB活性（TORC）的转换器。为了确定在应激过程中TORC2调节CRH转录的生理重要性，我们检查了TORC2在CRH神经元中的定位，以及CRH异质核（hn）RNA的变化，TORC2的核易位以及TORC2与CRH的结合之间的关系。启动子。免疫组织化学揭示了下丘脑室旁核（PVN）的背外侧（核细胞）和背囊（小细胞）区域的TORC2免疫反应性（irTORC2）。尽管在基础条件下染色大部分是胞质染色，但在约束30分钟后，背核区域的核irTORC2明显增加，并伴随CRH hnRNA水平增加。应激后4小时核irTORC2和CRH hnRNA水平恢复到基础水平。双重染色免疫组化显示100％的检测到的CRH神经元存在TORC2共同染色，约束30分钟后61％发生了核易位。 TORC2在背外侧PVN中的细胞分布不受约束的影响。染色质免疫沉淀实验显示，约束30分钟后，CRH启动子募集了TORC2和磷酸化CREB（pCREB），但是在应激后4 h，只有pCREB与CRH启动子相关。在约束压力下TORC2易位到下丘脑CRH神经元核并与CRH启动子相互作用以及CRH转录激活的论证，为TORC2参与CRH转录的生理调节提供了有力的证据。

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