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Study on functional roles of a single neuron in animal behaviors by functional transplant of channel rhodopsin 2

视紫红质2通道功能移植研究单个神经元在动物行为中的功能作用

基本信息

批准号：
21570082
负责人：
NAGAHAMA Tatsumi
金额：
$ 3.08万
依托单位：
Toho University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2011
项目状态：
已结题

项目摘要

In the present experiments we attempted functional transplant of a channel rhodopsin 2(ChR2) isolated from Chlamydomonas into the identified MA neuron in Aplysia buccal ganglia, and explored the functional roles of the MA in the feeding neural circuit. The preparations were incubated at 20 oC for 3 days after injection of the ChR2 expression vector(pNEX-ChR2) into the cell body of MA. When the cell body region of the MA was illuminated with bright blue light(100W Xenon lamp), the depolarizing responses were recorded in most of the postsynaptic follower neurons. This result was inconsistent with the electrophysiological data where the MA connects many follower neurons with inhibitory chemical synapses in addition to the electrical synapses. ChR2 tends to be expressed over the whole cell membrane and the bright light stimulation will depolarize the whole cell. Then there may be a possibility that the conduction of action potentials produced near the cell body might be suppressed in the axonal region and we can observe the electrically coupled depolarization alone. We stimulated the small region on the MA cell body with the fine spotlight(10μm diameter) produced by the blue laser beam and could record the IPSPs in most of the followers then. In addition, the rhythmic bursting responses were produced in the MA or the other buccal neurons when the MA cell body was illuminated with the long-lasting bright light or the fine spotlight, suggesting the potentiality of the MA to produce such rhythmic responses. The present study will propose that unsuitable light stimulation will produce the responses different from the living phenomena in the functional transplant of ChR2.

在本实验中，我们尝试了功能移植通道视紫红质2（ChR 2）从衣原体分离到确定MA神经元在Astrasia颊神经节，并探讨功能的功能作用的MA在摄食神经回路。在将ChR 2表达载体（pNEX-ChR 2）注射到MA细胞体中后，将制备物在20 ℃下孵育3天。当用100 W氙灯照射MA胞体区时，大多数突触后跟随神经元都记录到去极化反应。这一结果与电生理学数据不一致，其中MA将许多跟随神经元与除了电突触之外的抑制性化学突触连接。ChR 2倾向于在整个细胞膜上表达，并且强光刺激将使整个细胞去活化。然后，可能有一种可能性，即细胞体附近产生的动作电位的传导可能在轴突区域受到抑制，我们可以单独观察到电耦合去极化。我们用蓝色激光束产生的细光斑（直径10μm）刺激MA细胞体上的小区域，可以记录到大多数跟随者中的IPSP。此外，当MA细胞体被长时间强光或细聚光灯照射时，MA或其他颊神经元产生节律性爆发反应，表明MA具有产生这种节律性反应的潜力。本研究将提出，在ChR 2功能移植中，不适当的光刺激将产生不同于生命现象的反应。