Nishi M, Usuku T, Itose M, Fujikawa K, Hosokawa K, Matsuda K-I, Kawata M
Department of Anatomy and Neurobiology, Kyoto Prefectural University of Medicine, Kawaramachi Hirokoji, Kyoto 602-8566, Japan.
Neuroscience. 2007 Jun 8;146(4):1555-60. doi: 10.1016/j.neuroscience.2007.03.020. Epub 2007 Apr 30.
The hippocampal formation is a plastic brain structure important for certain types of learning and memory, and also vulnerable to the effects of stress and trauma. Since hippocampal neurons express high levels of corticosteroid receptor, the morphological changes, including alterations in the size of soma, and the length and number of neurites and spines, in response to glucocorticoids released as a result of stress are intriguing. In order to highlight the morphology of neurons that express glucocorticoid receptor (GR), we have generated a transgenic mouse line expressing green fluorescent protein (GFP) under the control of the GR promoter. We found strong green fluorescence in the pyramidal cell layer of the CA1 and CA2 regions and the granule cell layer of the dentate gyrus of the hippocampus in brain sections of the transgenic mice. GFP fluorescence was observed not only in somas, but also in neurites including both dendrites and axons. In dissociated culture, we also observed GFP fluorescence in the soma, neurites including both dendrites and axons, and dendritic spines. Microtubule-associated protein 2 immunopositive pyramidal-shaped neurons clearly showed two different populations, GFP positive and GFP negative neurons. These results indicate that this transgenic mouse line should be useful for live imaging of neuronal structure in animals as well as GR-positive cultured cells using GFP as a specific indicator.
海马结构是一种具有可塑性的脑结构,对某些类型的学习和记忆很重要,并且容易受到应激和创伤的影响。由于海马神经元表达高水平的皮质类固醇受体,因此对应激释放的糖皮质激素作出反应时的形态变化,包括胞体大小、神经突和棘的长度及数量的改变,都很引人关注。为了突出表达糖皮质激素受体(GR)的神经元的形态,我们构建了一种在GR启动子控制下表达绿色荧光蛋白(GFP)的转基因小鼠品系。我们在转基因小鼠脑切片的海马CA1和CA2区的锥体细胞层以及齿状回的颗粒细胞层中发现了强烈的绿色荧光。不仅在胞体中观察到了GFP荧光,在包括树突和轴突的神经突中也观察到了。在解离培养中,我们还在胞体、包括树突和轴突的神经突以及树突棘中观察到了GFP荧光。微管相关蛋白2免疫阳性的锥体形神经元明显显示出两种不同的群体,即GFP阳性和GFP阴性神经元。这些结果表明,这种转基因小鼠品系作为一种使用GFP作为特异性指示剂的工具,对于在动物体内对神经元结构以及GR阳性培养细胞进行实时成像应该是有用的。