Caltana Laura Romina, Heimrich Bernd, Brusco Alicia
Institute of Anatomy and Cell Biology, Center for Neuroscience, University of Freiburg, Freiburg, Germany.
Cell Biology and Neuroscience Institute, School of Medicine, University of Buenos Aires, Paraguay 2155, 3rd floor, Buenos Aires, 1114, Argentina.
J Mol Neurosci. 2015 Aug;56(4):773-781. doi: 10.1007/s12031-015-0499-4. Epub 2015 Feb 3.
Endocannabinoid receptors CB1R and CB2R are present in the CNS and modulate synaptic activity. By using an in vitro model, two concentrations of CB1R agonist ACEA at 0.5 and 5 μM doses and CB1R antagonist AM251 at 1 and 10 μM doses were administered in organotypic slice cultures of mouse hippocampus, and their effects on neurons and glial cells were analyzed at different time points. Exposure to low concentrations of ACEA (0.5 μM) did not seem to affect tissue organization, neuronal morphology, or glial response. In contrast, at a higher concentration of ACEA, many neurons in the dentate gyrus exhibited strong caspase-3 immunoreactivity. After treatment with AM251, we observed an increase in caspase-3 immunoreactivity and a downregulation of CB1R expression. Results show that long-term hippocampal slice cultures respond to both CB1R activation and inactivation by changing neuronal protein expression patterns. In the present study, we demonstrate that CB1R agonist ACEA promotes alterations in the neuronal cytoskeleton as well as changes in CB1R expression in organotypic hippocampal slice cultures, and that CB1R antagonist AM251 promotes neuronal death and astroglial reaction.
内源性大麻素受体CB1R和CB2R存在于中枢神经系统中并调节突触活动。通过使用体外模型，将两种浓度的CB1R激动剂ACEA（剂量分别为0.5和5μM）和CB1R拮抗剂AM251（剂量分别为1和10μM）施用于小鼠海马的器官型切片培养物中，并在不同时间点分析它们对神经元和神经胶质细胞的影响。暴露于低浓度的ACEA（0.5μM）似乎不会影响组织结构、神经元形态或神经胶质反应。相反，在较高浓度的ACEA作用下，齿状回中的许多神经元表现出强烈的caspase-3免疫反应性。用AM251处理后，我们观察到caspase-3免疫反应性增加以及CB1R表达下调。结果表明，长期海马切片培养物通过改变神经元蛋白表达模式对CB1R的激活和失活均有反应。在本研究中，我们证明CB1R激动剂ACEA促进器官型海马切片培养物中神经元细胞骨架的改变以及CB1R表达的变化，并且CB1R拮抗剂AM251促进神经元死亡和星形胶质细胞反应。
