利用成年小鼠脑片培养分析大脑区域特异性 mRNA 的合成和稳定性。

Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA.

Genetics and Aging Research Unit, MassGeneral Institute of Neurodegenerative Disease, Massachusetts General Hospital, Harvard Medical School, Boston, MA 02129, USA.

STAR Protoc. 2022 Apr 22;3(2):101349. doi: 10.1016/j.xpro.2022.101349. eCollection 2022 Jun 17.

Utilization of live animals for mechanistic study is challenging yet pivotal to elucidate pathogenesis of neurological diseases. Here, we present a protocol that employs cultured brain slices derived from adult mice to examine mRNA metabolism. We describe the preparation of acute brain slices and the treatments of RNA synthesis inhibitor and nucleotide analog to examine the effects of ataxin-1 loss-of-function on mRNA stability and transcription in cortex. This protocol also includes electrophysiological recording of spontaneous neuronal activity in hippocampus. For complete details on the use and execution of this protocol, please refer to Suh et al. (2019).

利用活体动物进行机制研究具有挑战性，但对于阐明神经疾病的发病机制至关重要。在这里，我们提出了一个使用从成年小鼠中培养的脑切片来研究 mRNA 代谢的方案。我们描述了急性脑切片的制备以及 RNA 合成抑制剂和核苷酸类似物的处理，以研究共济失调蛋白-1 功能丧失对皮质中 mRNA 稳定性和转录的影响。该方案还包括海马体中自发性神经元活动的电生理记录。有关此方案的使用和执行的完整详细信息，请参阅 Suh 等人。（2019 年）。