Development of neuronal subtypes and local circuits in the hippocampus

海马神经元亚型和局部回路的发育

基本信息

批准号：
10617334
负责人：
Jason C. Wester
金额：
$ 37.02万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-07-01 至 2026-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10617334
关键词：
ATAC-seq Acute Adolescent Adult Architecture Area Birth Brain Brain region Cell Differentiation process Cells Chromatin Chromatin Structure Classification Data Development Developmental Delay Disorders Disease Electric Stimulation Electrophysiology (science)Embryo Epigenetic Process Epilepsy Exhibits Experimental Designs Gene Expression Gene Expression Profiling Genetic Transcription Health Hippocampus Human Impairment Inhibitory Synapse Intellectual functioning disability Interneurons Knock-out Knowledge Learning Location Long-Term Potentiation Maintenance Maps Mediating Membrane Memory Modification Molecular Molecular Genetics Molecular Profiling Morphology Mus Mutant Strains Mice Mutation Neocortex Neuronal Differentiation Neurons Osteoblasts Output Physiology Play Positioning Attribute Process Property Proteins Pyramidal Cells Radial Regulator Genes Regulatory Pathway Role Schizophrenia Series Slice Source Specific qualifier value Structure Synapses Synaptic plasticity Techniques Testing Time Tissue-Specific Gene Expression Transcriptional Regulation Whole-Cell Recordings Work autism spectrum disorder autistic behaviour chromatin remodeling conditional knockout craniofacial disorder risk entorhinal cortex environmental enrichment for laboratory animals epigenetic regulation experience experimental study human disease insight memory consolidation migration neocortical neural neural circuit neurodevelopment neuron development novel strategies postnatal development recruit response risk variant single-cell RNA sequencing tool

项目摘要

PROJECT SUMMARY/ABSTRACT During brain development, neurons must properly differentiate into distinct subtypes to assemble healthy circuits. Thus, disruption of this process can impact neural architecture and wiring, and contribute to disorders such as autism, schizophrenia, and epilepsy. The hippocampus is a brain structure crucial for learning and memory, and its function is compromised in these disorders. Excitatory pyramidal cells in area CA1 provide a major output of hippocampal computations to other brain regions. These cells can be parsed based on their physical position within CA1 as “deep” or “superficial.” Deep and superficial hippocampal pyramidal cells are distinct classes of neurons that exhibit differential molecular signatures, electrophysiological properties, sources of afferent input, and circuit connectivity with local inhibitory interneurons. Determining the mechanisms underlying their differentiation is crucial for understanding hippocampal development and function in both health and disease. Superficial pyramidal cells in CA1 preferentially express the transcriptional regulator Satb2, which controls gene expression by modifying chromatin structure. In humans, mutations of Satb2 cause developmental delay, intellectual disability, epilepsy, and autistic behaviors. Our preliminary data show that knocking out Satb2 during early development in mice disrupts the differentiation of superficial pyramidal cells in CA1. Furthermore, there are non-cell-autonomous changes to the migration and survival of distinct subtypes of interneurons in mutant mice relative to controls. In the present proposal, three specific aims will test the hypothesis that early expression of Satb2 is necessary for hippocampal pyramidal cell differentiation and circuit development in CA1, while later expression is necessary to promote experience- dependent synaptic plasticity. These experiments will use molecular genetic tools in mice to conditionally knock out Satb2 from pyramidal cells during both early and late developmental stages. Aim 1 will use electrophysiology and electrical stimulation to study the strength and plasticity of different sources of afferent input to deep and superficial CA1 pyramidal cells in acute slices. This aim will test the hypothesis that early Satb2 expression is necessary to establish differences in afferent input strength, while later expression is necessary for activity-driven synaptic plasticity of these inputs. Aim 2 will use paired whole-cell recordings between pyramidal cells (deep and superficial) and identified subtypes of interneurons to map circuits and study details of their synaptic physiology. This aim will test the hypothesis that early Satb2 expression is necessary to establish circuit motifs between local inhibitory interneurons and superficial pyramidal cells, while later expression is necessary to recruit new inhibitory synapses in response to environmental enrichment. Aim 3 will use single-cell RNA-seq and ATAC-seq to determine how Satb2 knockout alters gene expression and chromatin accessibility in CA1 at multiple developmental timepoints. This aim will provide molecular insight into how Satb2 controls gene expression in CA1 through development, and how its function may change over time.

项目摘要/摘要在大脑发育过程中，神经元必须适当区分不同的亚型才能组装健康界。这一过程的破坏会影响神经架构和布线，并导致疾病例如自闭症，精神分裂症和癫痫。海马是对学习至关重要的大脑结构记忆及其功能在这些疾病中受到损害。 CA1区域中的兴奋性金字塔细胞提供海马计算对其他大脑区域的主要输出。这些细胞可以根据它们的 CA1中的物理位置为“深”或“表面”。深层和浅表海马锥体细胞是存在差异分子特征，电生理特性的不同类别的神经元，传入输入的来源以及与局部抑制性神经元的电路连接。确定其分化为基础的机制对于理解海马发展和功能至关重要在健康和疾病中。 CA1中的表面锥体细胞优先表达转录调节剂SATB2，它通过修饰染色质结构来控制基因表达。在人类中 SATB2导致发育延迟，智力残疾，癫痫和加速行为。我们的初步数据表明在小鼠早期发育过程中淘汰SATB2会破坏表面的分化 CA1中的锥体细胞。此外，对迁移和存活的非单元自主变化相对于对照组，突变小鼠中神经元的不同亚型。在本提案中，三个具体目标将检验以下假设：SATB2的早期表达对于海马锥体细胞是必需的 CA1中的分化和电路发展，而后来的表达是促进经验的必要表达 - 依赖的突触可塑性。这些实验将使用小鼠中的分子遗传工具进行有条件在早期和晚期发育阶段，从锥体细胞中敲出SATB2。 AIM 1将使用电生理学和电模拟研究不同来源的强度和可塑性输入急性切片中的深层和表面CA1锥体细胞。这个目标将检验以下假设 SATB2表达对于建立传入输入强度的差异是必要的，而以后的表达为这些输入的活动驱动的合成可塑性所必需的。 AIM 2将使用配对的全细胞录音在锥体细胞（深层和表面）之间，并确定了中间神经元的亚型以绘制电路和研究其突触生理学的细节。这个目的将检验以下假设：早期的SATB2表达是在局部抑制性神经元和浅表金字塔细胞之间建立电路基序所必需的，而以后的表达对于响应环境富集而募集新的抑制性突触是必要的。目的 3将使用单细胞RNA-seq和atac-seq来确定SATB2基因敲除如何改变基因表达和在多个发育时点，CA1中的染色质可及性。这个目标将为分子见解 SATB2如何通过开发控制CA1中的基因表达，以及其功能如何随时间变化。