Recording electrical activity from identified neurons in the intact brain of transgenic fish.
Recording electrical activity from identified neurons in the intact brain of transgenic fish.
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DOI：
10.3791/50312
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发表时间：
2013-04
期刊：
影响因子：
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通讯作者：
Yali Zhao;N. Wayne
中科院分区：
文献类型：
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作者：
Yali Zhao;N. Wayne
理解调节复杂行为的神经回路的细胞生理学通过使用模型系统大大增强，在该模型系统中，可以在CNS的神经回路保持完整的完整大脑准备中进行这项工作。我们使用转基因鱼，其中促性腺激素释放激素（GnRH）神经元的遗传标记与绿色荧光蛋白在完整的大脑中进行识别。鱼类有多个GnRH神经元群体，它们的功能取决于它们在大脑中的位置和它们表达的GnRH基因（1）。我们使用转基因青鳉鱼的完整脑集中展示了位于与嗅球相关的终末神经（TN）中的GnRH 3神经元（图1B和C）。研究表明青鳉TN-GnRH 3神经元具有神经调节作用，作为外界环境信息传递到中枢神经系统的传递者;它们在调节垂体-性腺功能中不起直接作用，如公知的下丘脑GnRH 1神经元那样（2，3）。TN-GnRH 3神经元自发动作电位放电的紧张模式是一种内在性质（4-6），其频率由来自同种的视觉提示（2）和神经肽kisspeptin 1（5）调节。在这个视频中，我们使用一个稳定的转基因青鳉品系，其中TN-GnRH 3神经元表达含有连接到增强型绿色荧光蛋白的GnRH 3启动子区域的转基因（7），向您展示如何识别神经元并监测其在整个大脑制备中的电活动（6）。
Understanding the cell physiology of neural circuits that regulate complex behaviors is greatly enhanced by using model systems in which this work can be performed in an intact brain preparation where the neural circuitry of the CNS remains intact. We use transgenic fish in which gonadotropin-releasing hormone (GnRH) neurons are genetically tagged with green fluorescent protein for identification in the intact brain. Fish have multiple populations of GnRH neurons, and their functions are dependent on their location in the brain and the GnRH gene that they express(1) . We have focused our demonstration on GnRH3 neurons located in the terminal nerves (TN) associated with the olfactory bulbs using the intact brain of transgenic medaka fish (Figure 1B and C). Studies suggest that medaka TN-GnRH3 neurons are neuromodulatory, acting as a transmitter of information from the external environment to the central nervous system; they do not play a direct role in regulating pituitary-gonadal functions, as do the well-known hypothalamic GnRH1 neurons(2, 3) .The tonic pattern of spontaneous action potential firing of TN-GnRH3 neurons is an intrinsic property(4-6), the frequency of which is modulated by visual cues from conspecifics(2) and the neuropeptide kisspeptin 1(5). In this video, we use a stable line of transgenic medaka in which TN-GnRH3 neurons express a transgene containing the promoter region of Gnrh3 linked to enhanced green fluorescent protein(7) to show you how to identify neurons and monitor their electrical activity in the whole brain preparation(6).