Wang Yubin, Baudry Michel
Graduate College of Biomedical Sciences, Western University of Health Sciences, Pomona, CA 91766, USA.
Bio Protoc. 2019 Mar 5;9(5):e3187. doi: 10.21769/BioProtoc.3187.
Acute cerebellar slices are widely used among neuroscientists to study the properties of excitatory and inhibitory synaptic transmission as well as intracellular signaling pathways involved in their regulation in cerebellum. The cerebellar cortex presents a well-organized circuitry, and several neuronal pathways can be stimulated and recorded reliably in acute cerebellar slices. A widely used acute cerebellar slice preparation technique was adapted from Edwards' thin slice preparation method published in 1989 ( Edwards , 1989 ). Most of the acute cerebellar slice preparation techniques use a vibrating microtome for slicing freshly dissected cerebellum from various animal species. Here we introduce a simpler method, which uses a tissue chopper to quickly prepare acute sagittal cerebellar slices from rodents. Cerebellum is dissected from the whole brain and sliced with a tissue chopper into 200-400 µm thick slices. Slices are allowed to recover in oxygenated aCSF at 37 °C for 1-2 h. Slices can then be used for electrophysiology or other types of experimentation. This method can be used to prepare cerebellar slices from mouse or rat aged from postnatal day 7 to 2 years. The preparation is faster and easier than other methods and provides a more versatile diversity of applications.
急性小脑切片在神经科学家当中被广泛用于研究兴奋性和抑制性突触传递的特性,以及小脑参与其调节的细胞内信号通路。小脑皮质呈现出组织良好的神经回路,并且在急性小脑切片中可以可靠地刺激和记录多条神经元通路。一种广泛使用的急性小脑切片制备技术改编自1989年发表的爱德华兹薄片制备方法(爱德华兹,1989年)。大多数急性小脑切片制备技术使用振动切片机对从各种动物物种新鲜解剖的小脑进行切片。在这里,我们介绍一种更简单的方法,该方法使用组织切片机从啮齿动物快速制备急性矢状小脑切片。从小脑整体中分离出小脑,并用组织切片机切成200 - 400微米厚的切片。切片在37℃的充氧人工脑脊液中恢复1 - 2小时。然后切片可用于电生理学或其他类型的实验。该方法可用于制备出生后第7天至2岁的小鼠或大鼠的小脑切片。该制备方法比其他方法更快、更容易,并且提供了更多样化的应用。