Investigating the role of the stress-responsive protein Tob in the brain

研究压力反应蛋白 Tob 在大脑中的作用

• 批准号: 18J20551
• 负责人: Youssef Mohieldin
• 金额: $ 1.79万
• 依托单位: Okinawa Institute of Science and Technology Graduate University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for JSPS Fellows
• 财政年份: 2018
• 资助国家: 日本
• 起止时间: 2018-04-25 至 2022-03-31
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18J20551/
• 关键词: Stress; Depression; Fear; Tob; Tob1; Anxiety; stress; anxiety; fear; brain

RNA sequencing analysis showed abnormal post-stress changes in the hippocampal transcriptome profile after Tob deletion that was validated by qRT-PCR for mRNA levels of altered genes. Pathway analysis suggested potential pathways and suggested interesting genes to further investigate.In order to understand the effect of Tob deletion on the single neuronal transcriptome level, I performed single-nucleus sequencing for samples from hippocampus of Tob wildtype (WT) and knockout (KO) mice. The aim was to characterize distinct subpopulation of neurons that underwent changes in the transcriptome that is correlated to the observed abnormal behavior after Tob deletion. The preparation of good quality samples from adult brain tissue was challenging but was eventually achieved after several optimization steps. Additionally, data analysis was not easy to set up in the beginning, but currently an analysis pipeline is set up and ready for any further analyses in the future. The results are currently undergoing reviewing to be used in future projects, despite the challenges encountered during the experimental procedure.Additionally, data generated in the past few years has been compiled, analyzed, and critically described in a manuscript, which has been submitted to an international scientific journal. The manuscript describes the function of Tob in the brain from behavior, circuit to molecular aspects in relation to the stress machinery.

RNA测序分析显示，ToB缺失后，应激后海马区转录组谱发生了异常变化，这一点通过qRT-PCR检测改变基因的mRNA水平得到了证实。为了了解Tob缺失对单个神经元转录组水平的影响，我对Tob野生型(WT)和基因敲除(KO)小鼠的海马区样本进行了单核测序。其目的是描述经历了转录组变化的神经元的不同亚群，这些变化与Tob缺失后观察到的异常行为有关。从成人脑组织中制备高质量的样本是具有挑战性的，但最终在几个优化步骤后实现了。此外，数据分析在开始时并不容易建立，但目前已经建立了分析管道，并为未来的任何进一步分析做好了准备。尽管在实验过程中遇到了挑战，但目前正在对结果进行审查，以用于未来的项目。此外，过去几年产生的数据已被汇编、分析并在已提交给一家国际科学期刊的手稿中进行了批判性描述。这篇手稿描述了ToB在大脑中的功能，从行为、回路到与应激机制有关的分子方面。

项目成果

Role of Transducer of ErbB2 (Tob) protein in the brain through regulating stress-induced behavior.

ErbB2 (Tob) 蛋白转导器通过调节应激诱导行为在大脑中的作用。

• 发表时间: 2021
• 作者: Mohieldin M. M. Youssef;Yuji Kiyama;Hiroaki Hamada;Esther Lai;Mohamed Eltabbal;Bernd Kuhn;Tadashi Yamamoto
• 通讯作者: Tadashi Yamamoto