Functional elucidation of m6A mRNA modification in spatial map formation in hippocampal CA1 pyramidal cells

海马 CA1 锥体细胞空间图形成中 m6A mRNA 修饰的功能阐明

• 批准号: 22K20702
• 负责人: Phasuk Sarayut
• 金额: $ 1.83万
• 依托单位: Institute of Physical and Chemical Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Research Activity Start-up
• 财政年份: 2022
• 资助国家: 日本
• 起止时间: 2022-08-31 至 2024-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-22K20702/
• 关键词: m6A; YTHDF3; YTHDF1; Place cells; Calcium Imaging; Two photon microscopy; Spatial memory; Virtual reality; Place Cells; Virtual Reality

We successfully generated conditional knockout (cKO) mouse models in which a m6A reader protein, YTHDF1 or YTHDF3 is deleted from the excitatory neurons. These mice are able to perform the task as shown by increasing running period during the habituation to non-virtual linear track task. We also found increase in running speed, licking number and pre licking behavior during the training sessions. Adeno-associated virus (AAV) containing GCaMP6f, a green calcium fluorescence sensor was introduced into hippocampal CA1 prior to brain imaging of mice performing VR task. This allowed us to observe the neuronal activity in hippocampal CA1 of the cKO mice while performing VR task.

我们成功地建立了条件基因敲除(CKO)小鼠模型，在该模型中，M6A阅读器蛋白YTHDF1或YTHDF3从兴奋性神经元中缺失。在对非虚拟直线跟踪任务的习服过程中，这些小鼠能够完成任务，表现为增加了运行时间。我们还发现，在训练过程中，跑步速度、舔食次数和预舔食行为都有所增加。将含有绿色钙荧光传感器GCaMP6f的腺相关病毒(AAV)引入海马CA1区，然后对进行VR任务的小鼠进行脑成像。这使得我们能够在执行VR任务的同时观察CKO小鼠海马CA1区神经元的活动。

项目成果

2-photon Calcium Imaging of Hippocampal Neuronal Circuits in Awake Head-Fixed Mice Navigating a Virtual Reality Environment

在虚拟现实环境中导航的清醒头部固定小鼠的海马神经元回路的 2 光子钙成像

• 发表时间: 2022
• 作者: Segi-Nishida Eri;Suzuki Kanzo;Sarayut Phasuk
• 通讯作者: Sarayut Phasuk