Whole cell recordings from brain of adult Drosophila.
Whole cell recordings from brain of adult Drosophila.
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DOI：
10.3791/248
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发表时间：
2007-01-01
期刊：
影响因子：
--
通讯作者：
O'Dowd, Diane K
中科院分区：
文献类型：
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作者：
Gu, Huaiyu;O'Dowd, Diane K
在这个视频中，我们演示了从成年果蝇中分离整个大脑的过程，为记录单个神经元做准备。我们开始描述解剖解决方案和捕获的成年女性在我们的研究中使用。完整切除整个大脑的过程，包括两个视叶，图示说明。上面气管的解剖也显示出来。分离的大脑不仅很小，而且在这个阶段处理时需要特别小心，以防止损伤神经元，其中许多神经元靠近组织的外表面。我们展示了我们开发的一种特殊的保持器是如何用于稳定记录室中的大脑的。标准电生理学设置用于记录单个神经元或成对神经元。通过记录显微镜观察的来自驱动GFP表达的GAL 4线（GH 146）的荧光图像说明了如何在活脑中鉴定投射神经元（PN）。高功率Nomarski图像显示了一个单一神经元的视图，该神经元正被作为全细胞记录的目标。当脑被成功地移除而没有损伤时，大多数神经元自发地活跃，激发动作电位和/或表现出自发的突触输入。这种原位制备，其中在整个大脑中识别的神经元的全细胞记录可以与遗传和药理学操作相结合，是一个有用的模型，用于探索成人CNS中的细胞生理学和可塑性。
In this video, we demonstrate the procedure for isolating whole brains from adult Drosophila in preparation for recording from single neurons. We begin by describing the dissecting solution and capture of the adult females used in our studies. The procedure for removing the whole brain intact, including both optic lobes, is illustrated. Dissection of the overlying trachea is also shown. The isolated brain is not only small but needs special care in handling at this stage to prevent damage to the neurons, many of which are close to the outer surface of the tissue. We show how a special holder we developed is used to stabilize the brain in the recording chamber. A standard electrophysiology set up is used for recording from single neurons or pairs of neurons. A fluorescent image, viewed through the recording microscope, from a GAL4 line driving GFP expression (GH146) illustrates how projection neurons (PNs) are identified in the live brain. A high power Nomarski image shows a view of a single neuron that is being targeted for whole cell recording. When the brain is successfully removed without damage, the majority of the neurons are spontaneously active, firing action potentials and/or exhibiting spontaneous synaptic input. This in situ preparation, in which whole cell recording of identified neurons in the whole brain can be combined with genetic and pharmacological manipulations, is a useful model for exploring cellular physiology and plasticity in the adult CNS.