Combining electrophysiology and optogenetics for functional screening of pyramidal neurons in the mouse prefrontal cortex

• Published in: STAR protocols 2021-06, Vol.2 (2), p.100469, Article 100469
• Main Authors: Nagahama, Kenichiro, Fujino, Shuhei, Watanabe, Takaki, Uesaka, Naofumi, Kano, Masanobu
• Format: Article
• Language: English
• Subjects: Animals; High Throughput Screening; Mice; Microscopy; Neural Pathways - metabolism; Neuroscience; Optogenetics; Prefrontal Cortex - metabolism; Protocol; Pyramidal Cells - metabolism; Synaptic Transmission
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2021.100469

Here, we present a comprehensive protocol to analyze the roles of disease-related genes in synaptic transmission. We have developed a pipeline of electrophysiological techniques and combined these with optogenetics in the medial prefrontal cortex of mice. This methodology provides a cost-effective, faster, and easier screening approach to elucidate functional aspects of single genes in several regions in the mouse brain such as a specific layer of the mPFC. For complete details on the use and execution of this protocol, please refer to Nagahama et al. (2020) and Sacai et al. (2020). [Display omitted] •Protocol to investigate roles of single genes in synaptic transmission in the mPFC•Construction and validation of miRNAs for KD of target single genes•In utero transfection of KD-miRNAs into subsets of pyramidal neurons in the mPFC•Functional and morphological analyses of synapses in mPFC neurons with or without KD Here, we present a comprehensive protocol to analyze the roles of disease-related genes in synaptic transmission. We have developed a pipeline of electrophysiological techniques and combined these with optogenetics in the medial prefrontal cortex of mice. This methodology provides a cost-effective, faster, and easier screening approach to elucidate functional aspects of single genes in several regions in the mouse brain such as a specific layer of the mPFC.