Biological characterization and optical imaging of brain-derived neurotrophic factor-green fluorescent protein suggest an activity-dependent local release of brain-derived neurotrophic factor in neurites of cultured hippocampal neurons
Biological characterization and optical imaging of brain-derived neurotrophic factor-green fluorescent protein suggest an activity-dependent local release of brain-derived neurotrophic factor in neurites of cultured hippocampal neurons
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DOI：
10.1002/jnr.1080
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发表时间：
2001-04-01
影响因子：
4.2
通讯作者：
Hatanaka, H
中科院分区：
文献类型：
--
作者：
Kojima, M;Takei, N;Hatanaka, H
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) 模仿了未融合（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，我们观察到该蛋白在成熟神经元的神经突中形成引人注目的簇，并与 PSD-95 免疫反应性共定位。共焦显微镜研究表明，这种聚集的 BDNF-GFP 在 KCl 去极化反应中迅速消失。这些数据表明 BDNF 以活动依赖性方式在突触位点局部快速释放。使用 BDNF-GFP 进行的光学研究可能为 BDNF 参与突触可塑性提供重要证据。 (C) 2001 Wiley-Liss, Inc.
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. (C) 2001 Wiley-Liss, Inc.