Project 1

项目1

• 批准号: 9924668
• 负责人: Takao K Hensch
• 金额: $ 38.49万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9924668
• 关键词: 3-Dimensional; Acoustics; Address; Adult; Age; Anatomy; Anxiety; Area; Attention; Behavior; Behavioral; Behavioral Paradigm; Biological; Biological Assay; Boston; Brain; Breathing; Calcium Chloride; Callithrix; Cell Maturation; Cell physiology; Cells; Chlorides; Cognition; Cognitive; Development; Dissection; Electrodes; Electroencephalography; Electrophysiology (science); Engineering; Equilibrium; Evolution; Exposure to; Foundations; Gait abnormality; Genes; Genetic; Grooming; Hindlimb; Human; Image; Impairment; Injections; Insula of Reil; Knock-out; Learning; Life; Litter Size; Locomotion; Longevity; Longitudinal Studies; Maps; Measures; Mental disorders; Methyl-CpG-Binding Protein 2; Modeling; Motor; Mus; Music; Mutant Strains Mice; Mutation; Neonatal; Neurologic; Neurologic Symptoms; Neurons; Organoids; Oxidation-Reduction; Parvalbumins; Patients; Pediatric Hospitals; Phase; Phenotype; Physiological; Physiology; Play; Prefrontal Cortex; Pregnancy; Primates; Recovery; Reversal Learning; Risk Factors; Role; Sampling; Sensory; Silicon; Stimulus; Structure; Symptoms; Tamoxifen; Testing; Time; Ultrasonics; Viral; Virus; Visual Cortex; Visual attention; Weight; Work; autism spectrum disorder; base; behavior test; circadian pacemaker; cognitive development; critical period; deprivation; designer receptors exclusively activated by designer drugs; early experience; experience; flexibility; gamma-Aminobutyric Acid; gene environment interaction; gene function; in vivo; inhibitory neuron; insight; multisensory; mutant; novel; preference; premature; reconstruction; restoration; risk variant; sensor; sensory cortex; sensory system; standard measure; tool; touchscreen; two-photon; vocalization; 3-Dimensional; Acoustics; Address; Adult; Age; Anatomy; Anxiety; Area; Attention; Behavior; Behavioral; Behavioral Paradigm; Biological; Biological Assay; Boston; Brain; Breathing; Calcium Chloride; Callithrix; Cell Maturation; Cell physiology; Cells; Chlorides; Cognition; Cognitive; Development; Dissection; Electrodes; Electroencephalography; Electrophysiology (science); Engineering; Equilibrium; Evolution; Exposure to; Foundations; Gait abnormality; Genes; Genetic; Grooming; Hindlimb; Human; Image; Impairment; Injections; Insula of Reil; Knock-out; Learning; Life; Litter Size; Locomotion; Longevity; Longitudinal Studies; Maps; Measures; Mental disorders; Methyl-CpG-Binding Protein 2; Modeling; Motor; Mus; Music; Mutant Strains Mice; Mutation; Neonatal; Neurologic; Neurologic Symptoms; Neurons; Organoids; Oxidation-Reduction; Parvalbumins; Patients; Pediatric Hospitals; Phase; Phenotype; Physiological; Physiology; Play; Prefrontal Cortex; Pregnancy; Primates; Recovery; Reversal Learning; Risk Factors; Role; Sampling; Sensory; Silicon; Stimulus; Structure; Symptoms; Tamoxifen; Testing; Time; Ultrasonics; Viral; Virus; Visual Cortex; Visual attention; Weight; Work; autism spectrum disorder; base; behavior test; circadian pacemaker; cognitive development; critical period; deprivation; designer receptors exclusively activated by designer drugs; early experience; experience; flexibility; gamma-Aminobutyric Acid; gene environment interaction; gene function; in vivo; inhibitory neuron; insight; multisensory; mutant; novel; preference; premature; reconstruction; restoration; risk variant; sensor; sensory cortex; sensory system; standard measure; tool; touchscreen; two-photon; vocalization

Abstract Mice – with their short gestation time, lifespan, large litter size, and above all, genetic tractability –are powerful experimental tools with which to explore mammalian brain development. One striking example is the reversible disruption of autism risk genes, like Mecp2 or Shank3. Switching on these genes after the emergence of fullblown phenotypes rescues several motor- related symptoms, including inertia, abnormal gait, weight, irregular breathing, repetitive grooming or hind-limb clasping. Instead, other neurological aspects are uncorrected in adulthood, such as anxiety or motor coordination. Of central relevance to human patients, it remains unknown to what extent higher cognition is impaired in Shank3 or Mecp2 mutant mice and whether recovery would also be limited to a critical period. The Hensch project will directly address sensitive periods and reversibility of dorsomedial prefrontal cortical (dmPFC) functions using novel behavioral tests of attention, cognitive flexibility and acoustic preference along with associated physiological measures from relevant areas. Pioneering work in the first phase of the Conte Center established the pivotal role of particular inhibitory neurons underlying critical period timing in mouse sensory systems. Parvalbumin (PV+) GABA circuit maturation dictates both the onset and closure of these windows of circuit refinement. Manipulations of psychiatric risk factors, such as circadian Clock gene disruption or redox dysregulation, can delay or extend developmental trajectories by upsetting the vulnerable PV+ component of local circuit excitatory-inhibitory balance. Recently, Hensch and Feng extended this principle to higher-order multi-sensory integration (MSI, commonly impaired in patients with autism) in the insular cortex. Shared MSI impairments in mice lacking either Shank3 (weak PV+) or Mecp2 (excessive PV+) suggest an optimal range of PV+ network function enables proper pruning of connections in the insula. Here, we will examine circuit physiology and anatomy before/after restoration of Shank3 or Mecp2 in mice, ultimately by full 3D EM circuit reconstruction with Lichtman also in marmosets carrying the same Shank3 deletion (from Feng). Further, our touchscreen two- choice visual attention assay, a multiple-choice foraging task to assess flexible rule learning, and preference for acoustic stimuli (music, ultrasonic calls) experienced early in life will probe dmPFC function in mutant and rescued mice. Electrophysiological recording and two-photon Calcium / Chloride imaging from the dmPFC in vivo will focus on PV+ networks in these areas across development, starting with comparison to Arlotta’s human organoids. Based on these many insights from mice, the impact of silencing/activating PV+ circuits in corresponding frontal cortical regions of PV-Cre marmosets (by Feng) using focal injections of viral DREADD constructs can be tested on analogous primate tasks of attention, cognitive flexibility and preference behavior at MIT. Our collective work will determine whether biological determinants of critical periods in sensory systems play a similar role in cognition, the corresponding circuit changes which are corrected when Shank3/Mecp2 symptoms are reversed and how much the mouse and primate brain differ.

抽象的 小鼠 - 妊娠时间短，寿命，大垃圾大小，最重要的是遗传性障碍性 - 是探索哺乳动物大脑发育的强大实验工具。一 引人注目的例子是自闭症风险基因（如MECP2或Shank3）的可逆破坏。 全面表型出现后打开这些基因会反应几种电动机 相关症状，包括惯性，抓地力异常，体重，不规则呼吸，重复性 修饰或后绑limb弯曲。相反，其他神经学方面未经纠正 成年，例如动画或运动协调。与人类患者的中心相关性 在Shank3或MECP2突变小鼠中，较高的认知在多大程度上尚不清楚。 以及恢复是否也仅限于关键时期。 Hensch项目将直接 解决背侧前额叶皮质（DMPFC）功能的敏感周期和可逆性 使用新颖的关注行为测试，认知灵活性和声学偏好以及 相关领域的相关物理措施。在第一阶段的开创性工作 孔戴中心确立了关键基础的特定抑制神经元的关键作用 鼠标感官系统中的周期定时。 Parvalbumin（PV+）GABA电路成熟 这些电路细化窗口的发作和闭合。精神科操作 风险因素，例如昼夜节律基因破坏或氧化还原失调，可能会延迟或扩展 通过破坏本地电路的脆弱的PV+组件来发育轨迹 兴奋性抑制平衡。最近，Hensch和Feng将此原理扩展到了高阶 岛状皮质中的多感官整合（MSI，通常在自闭症患者中受损）。 缺乏shank3（弱PV+）或MECP2（过度PV+）的小鼠中的MSI障碍 建议最佳的PV+网络功能范围可以适当修剪连接 岛。在这里，我们将在恢复Shank3之前/之后检查电路生理和解剖学 或MICE中的MECP2，最终通过Lichtman的完整3D EM电路重建也 带有相同shank3删除（从冯）的摩尔果会。此外，我们的触摸屏两种 选择视觉注意评估，一项多项选择觅食任务，以评估灵活规则学习， 对声学刺激（音乐，超声通话）的偏爱将证明 DMPFC在突变体中的功能并救出小鼠。电生理记录和两光子 来自DMPFC体内DMPFC的钙 /氯化物成像将集中在这些区域的PV+网络上 在开发过程中，从与Arlotta的人类器官进行比较开始。基于这些 来自小鼠的许多见解，沉默/激活PV+电路在相应的额叶中的影响 PV-CRE Marmosets的皮质区域（通过冯）使用病毒dreadd的焦点注射 可以根据类似的灵长类动物的注意，认知灵活性和 麻省理工学院的偏好行为。我们的集体工作将决定生物决定者是否 感觉系统中的关键时期在认知中起着相似的作用，相应的电路 Shank3/MECP2症状被逆转时纠正的变化以及多少 鼠标和主要大脑不同。