An optimized surgical approach for obtaining stable extracellular single-unit recordings from the cerebellum of head-fixed behaving mice.
An optimized surgical approach for obtaining stable extracellular single-unit recordings from the cerebellum of head-fixed behaving mice.
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DOI：
10.1016/j.jneumeth.2016.01.010
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发表时间：
2016-03-15
影响因子：
3
通讯作者：
Sillitoe RV
中科院分区：
文献类型：
--
作者：
White JJ;Lin T;Brown AM;Arancillo M;Lackey EP;Stay TL;Sillitoe RV
电生理记录方法对于了解大脑功能至关重要。在这些方法中，有各种执行单个单元记录的方法。然而，在体内记录单个单元时要克服的主要障碍是稳定性。稳定性差导致低信噪比，这使得分离神经元信号具有挑战性。需要适当的隔离来区分来自相邻细胞的信号或电生理学固有的噪声。隔离不足使得无法分析完整的动作电位波形。不稳定的一个常见原因是手术不充分。手术过程中出现的问题会导致失血、组织损伤和周围组织愈合不良，限制进入目标脑区，重要的是，固定小鼠头部的固定点不可靠。我们描述了一个优化的手术过程，确保有限的组织损伤，并描绘了一种方法，植入头板，以保持动物牢牢地在适当的位置。使用小脑作为模型，我们实现了一个细胞外记录技术，以获得行为小鼠浦肯野细胞和小脑核神经元的单个单位。我们通过在注射强有力的震颤性药物后保持单个单位来验证我们的方法的稳定性。我们在记录后进行了多次结构分析。我们的方法是研究活跃小鼠神经元功能的理想方法，并且对于记录不可避免的大量运动时的单神经元活动很有价值。我们提出的进入小脑的手术原则可以很容易地适用于检查其他脑区神经元的功能。
Electrophysiological recording approaches are essential for understanding brain function. Among these approaches are various methods of performing single-unit recordings. However, a major hurdle to overcome when recording single units in vivo is stability. Poor stability results in a low signal-to-noise ratio, which makes it challenging to isolate neuronal signals. Proper isolation is needed for differentiating a signal from neighboring cells or the noise inherent to electrophysiology. Insufficient isolation makes it impossible to analyze full action potential waveforms. A common source of instability is an inadequate surgery. Problems during surgery cause blood loss, tissue damage and poor healing of the surrounding tissue, limited access to the target brain region, and, importantly, unreliable fixation points for holding the mouse’s head. We describe an optimized surgical procedure that ensures limited tissue damage and delineate a method for implanting head plates to hold the animal firmly in place. Using the cerebellum as a model, we implement an extracellular recording technique to acquire single units from Purkinje cells and cerebellar nuclear neurons in behaving mice. We validate the stability of our method by holding single units after injecting the powerful tremorgenic drug harmaline. We performed multiple structural analyses after recording. Our approach is ideal for studying neuronal function in active mice and valuable for recording single-neuron activity when considerable motion is unavoidable. The surgical principles we present for accessing the cerebellum can be easily adapted to examine the function of neurons in other brain regions.