Head-anchored whole-cell recordings in freely moving rats
Head-anchored whole-cell recordings in freely moving rats
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DOI：
10.1038/nprot.2009.5
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发表时间：
2009-01-01
期刊：
影响因子：
14.8
通讯作者：
Brecht, Michael
中科院分区：
文献类型：
--
作者：
Lee, Albert K.;Epsztein, Jerome;Brecht, Michael
细胞内记录通常用于研究体外神经元的突触和内在特性。这些记录的一个关键要求是机械上非常稳定的准备;因此它们在体内的使用以前仅限于头部受限的动物。我们最近已经证明，将电极牢固地固定在相对于颅骨的适当位置，可以为清醒、自由移动的大鼠提供足够的稳定性，从而实现持久、高质量的全细胞记录。该协议详细描述了我们的程序，增加了针对海马CA 1锥体神经元的具体说明，并更新了有助于修补和提高成功率的变化。这些变化包括将标准的非头戴式显微操作器与夹具相结合，以便在锚定过程中牢固地固定记录移液管，然后轻轻地释放它。从手术开始到记录结束的过程大约需要5小时。该技术允许对自然行为期间识别的细胞中的神经元整合和细胞/突触可塑性的潜在机制进行新的研究。
Intracellular recordings are routinely used to study the synaptic and intrinsic properties of neurons in vitro. A key requirement for these recordings is a mechanically very stable preparation; thus their use in vivo had been limited previously to head-restrained animals. We have recently demonstrated that anchoring the electrode rigidly in place with respect to the skull provides sufficient stabilization for long-lasting, high-quality whole-cell recordings in awake, freely moving rats. This protocol describes our procedure in detail, adds specific instructions for targeting hippocampal CA1 pyramidal neurons and updates it with changes that facilitate patching and improve the success rate. The changes involve combining a standard, nonhead-mounted micromanipulator with a gripper to firmly hold the recording pipette during the anchoring process then gently release it afterwards. The procedure from the beginning of surgery to the end of a recording takes similar to 5 h. This technique allows new studies of the mechanisms underlying neuronal integration and cellular/synaptic plasticity in identified cells during natural behaviors.