Department of Anatomy and Cell Biology, College of Veterinary Medicine, and Research Institute for Veterinary Science, Seoul National University, Seoul, South Korea.
Department of Biochemistry and Molecular Biology, College of Dentistry, Research Institute of Oral Sciences, Gangneung-Wonju National University, Gangneung, South Korea.
Brain Behav. 2019 Sep;9(9):e01388. doi: 10.1002/brb3.1388. Epub 2019 Aug 20.
The aim of this study was to investigate the effects of cuprizone on adult hippocampal neurogenesis in naïve mice. Additionally, we also studied how melatonin affects the neuronal degeneration induced by cuprizone.
Eight-week-old male C57BL/6J mice were randomly divided into three groups: (a) the control group, (b) the group treated with cuprizone only, and (c) the group treated with both cuprizone and melatonin. Cuprizone was administered with food at 0.2% ad libitum for 6 weeks. Melatonin was also administered with tap water at 6 g/L ad libitum for 6 weeks; the animals were then euthanized for immunohistochemistry with Ki67, doublecortin (DCX), glucose transporter 3 (GLUT3), and phosphorylation of cyclic adenosine monophosphate (AMP) response element binding (pCREB); double immunofluorescence of neuronal nuclei (NeuN) and myelin basic protein (MBP); and Western blot analysis of brain-derived neurotrophic factor (BDNF) expression to reveal the effects of cuprizone and melatonin on cell damage and hippocampal neurogenesis.
Administration of cuprizone significantly decreased the number of differentiating (DCX-positive) neuroblasts and proliferating (Ki67-positive) cells in the dentate gyrus. Moreover, cuprizone administration decreased glucose utilization (GLUT3-positive cells) and cell transcription (pCREB-positive cells and BDNF protein expression) in the dentate gyrus. Administration of melatonin ameliorated the cuprizone-induced reduction of differentiating neuroblasts and proliferating cells, glucose utilization, and cell transcription.
The results of the study suggest that cuprizone treatment disrupts hippocampal neurogenesis in the dentate gyrus by reducing BDNF levels and decreasing the phosphorylation of CREB. These effects were ameliorated by melatonin treatment.
本研究旨在探讨双硫仑对新生小鼠海马神经发生的影响。此外，我们还研究了褪黑素如何影响双硫仑诱导的神经元变性。
将 8 周龄雄性 C57BL/6J 小鼠随机分为三组：（a）对照组，（b）仅用双硫仑处理组，（c）用双硫仑和褪黑素处理组。双硫仑以 0.2%的浓度添加到食物中自由摄取 6 周。褪黑素也以 6g/L 的浓度添加到自来水中自由摄取 6 周；然后处死动物进行免疫组织化学检测，用 Ki67、双皮质素（DCX）、葡萄糖转运蛋白 3（GLUT3）和环磷酸腺苷（AMP）反应元件结合蛋白（pCREB）磷酸化（pCREB）；神经元核（NeuN）和髓鞘碱性蛋白（MBP）的双重免疫荧光；以及脑源性神经营养因子（BDNF）表达的 Western blot 分析，以揭示双硫仑和褪黑素对细胞损伤和海马神经发生的影响。
双硫仑给药显著减少了齿状回中分化（DCX 阳性）神经前体细胞和增殖（Ki67 阳性）细胞的数量。此外，双硫仑给药减少了齿状回中的葡萄糖利用（GLUT3 阳性细胞）和细胞转录（pCREB 阳性细胞和 BDNF 蛋白表达）。褪黑素给药改善了双硫仑诱导的分化神经前体细胞和增殖细胞、葡萄糖利用和细胞转录减少。
研究结果表明，双硫仑通过降低 BDNF 水平和减少 CREB 的磷酸化来破坏齿状回中的海马神经发生。这些作用可被褪黑素治疗改善。
