Analysis of brain region-specific mRNA synthesis and stability by utilizing adult mouse brain slice culture

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

Abstract

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 Bace1 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).

Keywords: Cell Biology; Gene Expression; Genetics; Neuroscience.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amyloid Precursor Protein Secretases* / metabolism
  • Animals
  • Aspartic Acid Endopeptidases* / metabolism
  • Brain / metabolism
  • Mice
  • Neurons / metabolism
  • RNA, Messenger / genetics

Substances

  • RNA, Messenger
  • Amyloid Precursor Protein Secretases
  • Aspartic Acid Endopeptidases