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

RNA测序分析显示，Tob缺失后海马转录组谱在应激后发生异常变化，qRT-PCR证实了改变基因的mRNA水平。途径分析提示了潜在的途径和值得进一步研究的有趣基因。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）和敲除型（KO）小鼠海马样本进行了单核测序。目的是表征不同的神经元亚群，这些神经元在Tob缺失后经历了与观察到的异常行为相关的转录组变化。从成人脑组织中制备高质量样品具有挑战性，但经过几个优化步骤后最终实现了。此外，数据分析在一开始并不容易建立，但目前已经建立了一个分析管道，并为将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，以便在未来的项目中使用。此外，过去几年产生的数据已被汇编、分析，并在一份手稿中进行了批判性描述，该手稿已提交给一家国际科学期刊。本文从行为学、电路学、分子学等方面描述了Tob在大脑中的功能与应激机制的关系。

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.