Correlating physiology and transcription during learning in a model for the 22q11.2 deletion

项目摘要 已经证实，情节记忆(EM)的编码部分由海马区CA1区的位置细胞活动和调谐决定。然而，细胞促进这种学习能力的生物学机制在很大程度上仍然不清楚。精神分裂症(SCZ)是一种通常发生在成年早期的疾病，仅在美国就有大约1/100的人受到影响，并表现出严重的认知障碍，包括EM功能障碍。在目标导向的学习和记忆任务中，使用钙成像技术记录活体行为小鼠的位置细胞活动。研究表明，22q11.2缺失综合征(DF(16)A+/-)转基因小鼠模型中的位置细胞动力学受到损害。22q11.2缺失综合征是SCZ的最大遗传原因之一。然而，这些赤字背后的机制仍未被探索。这项建议的目的有两个：1)将定位细胞活动的生理变化与其转录变化相关联，以更好地了解海马中定位细胞调节的分子生物学；2)比较野生型和DF(16)A+/-小鼠体内转录组的变化，以揭示在EM编码过程中22q11.2缺失综合征模型中被破坏的生物学过程。 为了解决这一问题，将执行以下目标：目标1：WT小鼠海马区CA1区的大规模补丁和单细胞RNA测序。将对CA1区不同类型的细胞进行转录、电生理和形态表征，包括锥体细胞、中间神经元和神经胶质细胞类型。目的：研究WT和DF(16)A+/-小鼠海马CA1区生理及相关转录水平的变化。我将使用rAAV传递的遗传编码钙指示剂(GECI)将WT和突变小鼠的海马区细胞功能障碍与转录变化联系起来。在WT和DF(16)A+/-小鼠CA1PC中，在目标定向学习前后执行scRNAseq。目的3：通过单细胞操作纠正DF(16)A+/-小鼠CA1神经元或神经胶质细胞生理特性的改变。利用rAAV提供的GECI，我将在体内进行电穿孔(IVE)，以纠正AIM 2中识别的失调基因，并恢复DF(16)A+/-小鼠的正常生理功能。

Project Summary It has been established that episodic memory (EM) encoding is dictated in part by place cell activity and tuning in the CA1 region of the hippocampus. However, it remains largely unknown what the biological mechanisms in place cells are that facilitate this learning capacity. Schizophrenia (SCZ) is a disease that typically arises in early adulthood, affecting roughly 1 in 100 people in the United States alone and exhibit significant cognitive impairments including EM dysfunction. Using calcium imaging to record place cell activity in live behaving mice in a goal-directed learning and memory task it was shown that place cell dynamics in a transgenic mouse model for 22q11.2 deletion syndrome (Df(16)A+/- ), one of the largest known genetic causes of SCZ, are impaired. However, the mechanisms underlying these deficits are still unexplored. The purpose of this proposal is 2 fold: to 1) correlate physiological changes in place cell activity with their transcriptomic changes to better understand the molecular biology underlying place cell tuning in the hippocampus, and 2) to compare changes in the transcriptome in wildtype and Df(16)A+/- mice in vivo to uncover biological processes disrupted in a model for 22q11.2 deletion syndrome during EM encoding. To address this, the following Aims will be performed: Aim 1: Large-scale patch and single cell RNA sequencing of the hippocampal CA1 region in WT mice. Transcriptional, electrophysiological and morphological characterization will be performed on different cell types in the CA1 region including pyramidal, interneuron, and glial cell types. Aim 2: In vivo physiological and correlated transcriptomic profiling in the mouse hippocampal CA1 of WT and Df(16)A+/- mice. I will use genetically encoded calcium indicators (GECIs) delivered by rAAV to correlate hippocampal place cell dysfunction with transcriptomic changes in both WT and mutant mice. scRNAseq will be performed before and after goal-directed learning in WT and Df(16)A+/- mice CA1PCs. Aim 3: Single-cell manipulations to correct altered in vivo physiological properties of CA1 neurons or glia in Df(16)A+/- mice. Using GECIs delivered by rAAV, I will perform in vivo electroporation (IVE) to correct dysregulated genes identified from Aim 2 and restore normal physiological function in Df(16)A+/- mice.