用基因可定位的电压指示器监视神经膜潜力
Journal of the American Chemical Society
|January 11, 2019
概括
新的VoltageSpy染料可以精确地测量基因定义的神经元中的电压变化. 这些化学合成的光探针可以针对特定的细胞,为神经元活动成像提供高灵敏度和快速响应时间.
科学领域:
- 神经科学
- 生物化学
- 光学成像
背景情况:
- 与传统方法相比,光学电压测量提供了更好的空间和时间分辨率.
- 现有的电压敏感染料缺乏特定的细胞向能力,限制了它们的应用.
- 遗传编码的电压指示器提供细胞特异性,但通常具有较慢的动力学和较低的灵敏度.
研究的目的:
- 开发一种新型的电压敏感光染料 (sarcosine-VoltageFluors),可以对特定的细胞进行基因向.
- 在基因定义的神经元,包括轴突和树突中实现高分辨率的电压动态成像.
- 将合成染料的速度和灵敏度与基因探针的细胞特异性结合起来.
主要方法:
- 基于光素的功能性碳酸组的电压敏感光染料的合成.
- 将染料与聚乙烯糖醇 (PEG) 连接剂和SpyTag结合,以对SpyCatcher进行特定的结合.
- 在神经元的细胞表面表达SpyCatcher以提供向染料.
- 使用VoltageSpy系统在培养的神经元中的动作潜力的光学成像.
主要成果:
- 新的VoltageSpy染料有效标记细胞表达细胞表面SpyCatcher.
- VoltageSpy染料表现出良好的电压敏感性和快速反应动力学,超过了遗传编码的电压指标.
- 在神经元 soma,轴突和树突中实现了强大的单次试验光学检测.
- 在低纳米染料度下,电压成像是成功的.
结论:
- 它代表了电压成像技术的重大进步, 能够对神经元的电活动进行基因定向的光学测量.
- 这种技术结合了合成电压传感器和基因定位的优势,为神经元功能的详细研究铺平了道路.
- VoltageSpy为神经科学研究提供了一个强大的工具,
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