Huang Jingjing, Tufan Turan, Deng Maoxian, Wright Gary, Zhu Meng-Yang
State Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, China.
Departments of Biomedical Sciences, Quillen College of Medicine, East Tennessee State University, Johnson City, TN, USA.
J Neurochem. 2015 Oct;135(1):38-49. doi: 10.1111/jnc.13268. Epub 2015 Aug 18.
Corticotropin releasing factor (CRF) has been implicated to act as a neurotransmitter or modulator in central nervous activation during stress. In this study, we examined the regulatory effect of CRF on the expression and function of the norepinephrine transporter (NET) in vitro. SK-N-BE (2) M17 cells were exposed to different concentrations of CRF for different periods. Results showed that exposure of cells to CRF significantly increased mRNA and protein levels of NET in a concentration- and time-dependent manner. The CRF-induced increase in NET expression was mimicked by agonists of either CRF receptor 1 or 2. Furthermore, similar CRF treatments induced a parallel increase in the uptake of [(3) H] norepinephrine. Both increased expression and function of NET caused by CRF were abolished by simultaneous administration of CRF receptor antagonists, indicating a mediation by CRF receptors. However, there was no additive effect for the combination of both receptor antagonists. Chromatin immunoprecipitation assays confirm an increased acetylation of histone H3 on the NET promoter following treatment with CRF. Taken together, this study demonstrates that CRF up-regulates the expression and function of NET in vitro. This regulation is mediated through CRF receptors and an epigenetic mechanism related to histone acetylation may be involved. This CRF-induced regulation on NET expression and function may play a role in development of stress-related depression and anxiety. This study demonstrated that corticotropin release factor (CRF) up-regulated the expression and function of norepinephrine transporter (NET) in a concentration- and time-dependent manner, through activation of CRF receptors and possible histone acetylation in NET promoter. The results indicate that their interaction may play an important role in stress-related physiological and pathological status.
促肾上腺皮质激素释放因子（CRF）被认为在应激过程中作为神经递质或调节剂参与中枢神经激活。在本研究中，我们在体外检测了CRF对去甲肾上腺素转运体（NET）表达和功能的调节作用。将SK-N-BE (2) M17细胞暴露于不同浓度的CRF中不同时间。结果显示，细胞暴露于CRF后，NET的mRNA和蛋白水平显著增加，呈浓度和时间依赖性。CRF受体1或2的激动剂可模拟CRF诱导的NET表达增加。此外，类似的CRF处理导致[³H]去甲肾上腺素摄取平行增加。CRF受体拮抗剂同时给药可消除CRF引起的NET表达和功能增加，表明由CRF受体介导。然而，两种受体拮抗剂联合使用没有相加作用。染色质免疫沉淀试验证实，CRF处理后NET启动子上组蛋白H3的乙酰化增加。综上所述，本研究表明CRF在体外上调NET的表达和功能。这种调节通过CRF受体介导，可能涉及与组蛋白乙酰化相关的表观遗传机制。CRF诱导的对NET表达和功能的调节可能在应激相关的抑郁和焦虑的发生中起作用。本研究表明，促肾上腺皮质激素释放因子（CRF）通过激活CRF受体以及NET启动子中可能的组蛋白乙酰化，以浓度和时间依赖性方式上调去甲肾上腺素转运体（NET）的表达和功能。结果表明，它们的相互作用可能在应激相关的生理和病理状态中起重要作用。
