Instituto de Investigaciones Farmacológicas, Universidad de Buenos Aires - Consejo Nacional de Investigaciones Científicas y Técnicas, Junín 956, piso 5, Ciudad Autónoma de Buenos Aires, C1113, Buenos Aires, Argentina.
Department of Behavioral Sciences, San Diego Mesa College, San Diego, CA, USA.
Neurotox Res. 2020 Aug;38(2):498-507. doi: 10.1007/s12640-020-00200-5. Epub 2020 May 4.
Psychostimulant drugs, such as modafinil and caffeine, induce transcriptional alterations through the dysregulation of epigenetic mechanisms. We have previously demonstrated that acute modafinil administration is accompanied by multiple changes in the expression of histone deacetylases (HDACs) within the mouse medial prefrontal cortex (mPFC). Herein, we compared alterations in class IIa HDACs in the mouse mPFC and dorsal striatum (DS) after a single exposure to each psychostimulant. We treated male C57BL/6 mice with modafinil (90 mg/kg, i.p.), caffeine (10 mg/kg, i.p.), or vehicle and evaluated locomotor activity. Following, we examined hdac4, hdac5, and hdac7 mRNA expression using qRT-PCR and HDAC7, pHDAC7, and pHDACs4/5/7 using Western blot. Last, we explored generalized effects in N2a cell line using modafinil (100 μM and 1 mM) or caffeine (80 μM and 800 μM). Our results indicate that modafinil had greater effects on locomotor activity compared with caffeine. qRT-PCR experiments revealed that modafinil decreased hdac5 and hdac7 mRNA expression in the DS, while caffeine had no effects. In the mPFC, modafinil increased hdac7 mRNA expression, with no effects observed for caffeine. Western blot revealed that within the DS, modafinil induced increases in HDAC7, pHDAC7, and pHDACs4/5/7 protein expression, while, in the mPFC, caffeine induced decreases in HDAC7, pHDAC7, and pHDACs4/5/7 protein levels. In vitro studies revealed that modafinil increased hdac4, hdac5, and hdac7 mRNA levels in N2a, while caffeine only increased hdac5 at a higher dose. These findings support the notion that modafinil and caffeine exert distinct regulation of class IIa HDAC family members and that these transcriptional and translational consequences are region-specific.
精神兴奋剂药物,如莫达非尼和咖啡因,通过表观遗传机制的失调诱导转录改变。我们之前已经证明,急性莫达非尼给药伴随着小鼠内侧前额叶皮层(mPFC)中组蛋白去乙酰化酶(HDACs)表达的多种变化。在此,我们比较了单次暴露于每种精神兴奋剂后,小鼠 mPFC 和背侧纹状体(DS)中 IIa 类 HDAC 的变化。我们用莫达非尼(90mg/kg,ip)、咖啡因(10mg/kg,ip)或载体处理雄性 C57BL/6 小鼠,并评估了运动活性。随后,我们使用 qRT-PCR 检测了 hdac4、hdac5 和 hdac7 的 mRNA 表达,使用 Western blot 检测了 HDAC7、pHDAC7 和 pHDACs4/5/7。最后,我们使用莫达非尼(100μM 和 1mM)或咖啡因(80μM 和 800μM)在 N2a 细胞系中探索了一般影响。我们的结果表明,与咖啡因相比,莫达非尼对运动活性的影响更大。qRT-PCR 实验表明,莫达非尼降低了 DS 中的 hdac5 和 hdac7 mRNA 表达,而咖啡因没有影响。在 mPFC 中,莫达非尼增加了 hdac7 mRNA 的表达,而咖啡因则没有影响。Western blot 显示,在 DS 中,莫达非尼诱导 HDAC7、pHDAC7 和 pHDACs4/5/7 蛋白表达增加,而在 mPFC 中,咖啡因诱导 HDAC7、pHDAC7 和 pHDACs4/5/7 蛋白水平降低。体外研究表明,莫达非尼增加了 N2a 中的 hdac4、hdac5 和 hdac7 mRNA 水平,而咖啡因仅在更高剂量下增加了 hdac5。这些发现支持这样的观点,即莫达非尼和咖啡因对 IIa 类 HDAC 家族成员有不同的调节作用,这些转录和翻译的后果是区域特异性的。