Lee Doyun, Lee Albert K
Center for Cognition and Sociality, Institute for Basic Science, Daejeon 34141, Republic of Korea;
Howard Hughes Medical Institute, Janelia Research Campus, Ashburn, Virginia 20147
Cold Spring Harb Protoc. 2017 Apr 3;2017(4):pdb.prot095810. doi: 10.1101/pdb.prot095810.
Whole-cell recording is a key technique for investigating synaptic and cellular mechanisms underlying various brain functions. However, because of its high sensitivity to mechanical disturbances, applying the whole-cell recording method to freely moving animals has been challenging. Here, we describe a technique for obtaining such recordings in freely moving, drug-free animals with a high success rate. This technique involves three major steps: obtaining a whole-cell recording from awake head-fixed animals, reliable and efficient stabilization of the pipette with respect to the animal's head using an ultraviolet (UV)-transparent collar and UV-cured adhesive, and rapid release of the animal from head fixation without loss of the recording. This technique has been successfully applied to obtain intracellular recordings from the hippocampus of freely moving rats and mice exploring a spatial environment, and should be generally applicable to other brain areas in animals engaged in a variety of natural behaviors.
全细胞记录是研究各种脑功能背后的突触和细胞机制的关键技术。然而，由于其对机械干扰高度敏感，将全细胞记录方法应用于自由活动的动物一直具有挑战性。在此，我们描述了一种在无药物的自由活动动物中以高成功率获得此类记录的技术。该技术包括三个主要步骤：从清醒的头部固定动物获得全细胞记录，使用紫外线（UV）透明颈圈和UV固化粘合剂相对于动物头部可靠且高效地稳定移液管，以及在不丢失记录的情况下快速将动物从头部固定中释放出来。该技术已成功应用于从探索空间环境的自由活动大鼠和小鼠的海马体中获得细胞内记录，并且应该普遍适用于参与各种自然行为的动物的其他脑区。
