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

项目摘要 已经确定情景记忆（EM）编码部分由海马CA 1区的位置细胞活动和调谐决定。然而，在很大程度上仍然不清楚细胞中促进这种学习能力的生物学机制是什么。精神分裂症（SCZ）是一种通常在成年早期出现的疾病，仅在美国就影响大约1/100的人，并表现出显著的认知障碍，包括EM功能障碍。在目标导向的学习和记忆任务中，使用钙成像来记录行为正常的小鼠中的位置细胞活性，结果表明，22q11.2缺失综合征（Df（16）A+/-）的转基因小鼠模型中的位置细胞动力学受损，22q11.2缺失综合征是SCZ的最大已知遗传原因之一。然而，这些缺陷背后的机制仍然是未知的。该提议的目的有两个：1）将位置细胞活性的生理变化与其转录组学变化相关联，以更好地理解海马中位置细胞调谐的分子生物学基础，以及2）比较野生型和Df（16）A+/-小鼠体内转录组的变化，以揭示EM编码期间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，将进行以下目标：目标1：WT小鼠海马CA 1区的大规模贴片和单细胞RNA测序。将对CA 1区的不同细胞类型（包括锥体细胞、中间神经元和神经胶质细胞类型）进行转录、电生理和形态学表征。目的2：WT和Df（16）A+/-小鼠海马CA 1区的体内生理和相关转录组学分析。我将使用基因编码的钙指标（GECIs）提供的rAAV相关的海马位置细胞功能障碍与转录组的变化在WT和突变小鼠。scRNAseq将在WT和Df（16）A+/-小鼠CA 1 PC中的目标导向学习之前和之后进行。目的3：单细胞操作以纠正Df（16）A+/-小鼠CA 1神经元或神经胶质细胞体内生理特性的改变。使用由rAAV递送的GECIs，我将进行体内电穿孔（IVE）以校正从Aim 2鉴定的失调基因并恢复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.