EAGER: Targeted disruption of neural processes in zebrafish using femtosecond laser pulses to dissect the functional anatomy of the nervous system

1050134 Schaffer阐明系统某一部分的功能的一种方法是扰乱和破坏该部分并观察其效果。例如，通过对敲除和敲入转基因动物的研究，这种方法利用分子生物学工具，使人们更好地了解了许多疾病的遗传起源。虽然这种基因操作技术非常先进，但物理破坏技术，特别是非常精细或难以获得结构的技术，仍然很粗糙。能够以亚微米的空间精度破坏特定靶向结构的技术的发展，在没有重大附带影响的情况下，以及在自然的活体环境中，将为各种研究打开大门，这些研究可以与转基因实验的影响相匹配。飞秒激光脉冲具有独特的能力，可以在不影响材料表面的情况下，将能量沉积到透明材料的主体中的微观体积中。PI建议开发和使用这种能力来破坏活体动物神经系统中的特定靶向结构，目的是阐明功能。特别是，通过切开斑马鱼后脑的单个轴突和树突来解剖神经回路，研究行为的神经基础。

1050134SchafferOne way to elucidate the function of part of a system is to disrupt and break that part and observe the effect. Using molecular biology tools, this method has led to a better understanding of the genetic origins of many diseases through studies of knock-out and knock-in transgenic animals, for example. While such genetic manipulation technologies are very advanced, techniques for physical disruption, especially of very fine-scale or difficult to access structures, remain crude. Development of technologies that allow specifically targeted structures to be disrupted with sub-micrometer spatial precision, without significant collateral effects, and in native, in vivo environments would open the door to a variety of studies that could match the impact transgenic experiments have had. Femtosecond laser pulses have the unique capability to deposit energy into a microscopic volume in the bulk of a transparent material without affecting the surface of the material. The PI proposes to develop and use this capability to disrupt specifically targeted structures in the nervous system of live animals with the goal of elucidating function. In particular, to study the neural basis of behavior by dissecting neural circuits through cuts to individual axons and dendrites in zebrafish hindbrain.