Kojima M, Takei N, Numakawa T, Ishikawa Y, Suzuki S, Matsumoto T, Katoh-Semba R, Nawa H, Hatanaka H
Precursory Research for Embryonic Science and Technology, Japan Science and Technology Cooperation, Kawaguchi, Saitama, Japan.
J Neurosci Res. 2001 Apr 1;64(1):1-10. doi: 10.1002/jnr.1080.
To visualize the release dynamics of the brain-derived neurotrophic factor (BDNF) involved in neural plasticity, we constructed a plasmid encoding green fluorescent protein (GFP) fused with BDNF. First, several biological studies confirmed that this fusion protein (BDNF-GFP) mimics the biological functions and the release kinetics of unfused (native) BDNF. Second, when BDNF-GFP was expressed in cultured hippocampal neurons, we observed that this protein formed striking clusters in the neurites of mature neurons and colocalized with the PSD-95 immunoreactivity. Such a clustered BDNF-GFP rapidly disappeared in response to depolarization with KCl, as revealed by confocal microscopic studies. These data suggest that BDNF is locally and rapidly released at synaptic sites in an activity-dependent manner. Optical studies using BDNF-GFP may provide important evidence regarding the participation of BDNF in synaptic plasticity.
为了可视化参与神经可塑性的脑源性神经营养因子（BDNF）的释放动态，我们构建了一个编码与BDNF融合的绿色荧光蛋白（GFP）的质粒。首先，多项生物学研究证实，这种融合蛋白（BDNF-GFP）模拟了未融合（天然）BDNF的生物学功能和释放动力学。其次，当BDNF-GFP在培养的海马神经元中表达时，我们观察到这种蛋白在成熟神经元的神经突中形成了明显的簇，并与PSD-95免疫反应性共定位。共聚焦显微镜研究显示，这种聚集的BDNF-GFP在KCl去极化后迅速消失。这些数据表明，BDNF以活动依赖的方式在突触部位局部快速释放。使用BDNF-GFP的光学研究可能为BDNF参与突触可塑性提供重要证据。
