Prefrontal function in the Shank3-deficient rat: A first rat model for ASD

Shank3 缺陷大鼠的前额叶功能：第一个自闭症谱系障碍 (ASD) 大鼠模型

• 批准号: 8880287
• 负责人: Joseph D. Buxbaum
• 金额: $ 45.79万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-07-01 至 2019-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8880287
• 关键词: 3-Dimensional; Affect; Animal Model; Animals; Area; Attention; Attentional deficit; Autistic Disorder; Beds; Behavior; Behavior assessment; Behavioral; Biochemical; Biological Assay; Biological Models; Brain region; Cognitive; Communication; Development; Developmental Delay Disorders; Disease; Disease model; Drug Industry; Drug Kinetics; Eating; Electrons; Experimental Models; Genes; Genetic Engineering; Glutamates; Goals; Health; Hippocampus (Brain); Human; Impaired cognition; In Vitro; Intellectual functioning disability; Investigation; Lead; Learning; Length; Link; Medial; Methods; Mission; Modeling; Molecular; Molecular Target; Morphology; Motor; Mus; Neurobiology; Neurodevelopmental Disorder; Neurons; Neuropsychology; Outcome; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phenotype; Physiological; Physiology; Plastics; Prefrontal Cortex; Property; Proteins; Public Health; Rattus; Research; Resolution; Reversal Learning; Rodent Model; Role; Slice; Social Behavior; Social Functioning; Speech Delay; Structure; Symptoms; Synapses; Synaptic Receptors; Syndrome; System; Techniques; Testing; Three-Dimensional Imaging; Three-dimensional analysis; United States National Institutes of Health; Vertebral column; Work; Zinc Fingers; autism spectrum disorder; base; behavioral outcome; behavioral study; cognitive function; density; drug development; feeding; flexibility; in vivo; innovation; memory recognition; mouse model; neurochemistry; neurophysiology; neurotransmission; new therapeutic target; novel; novel therapeutics; nuclease; postsynaptic; pre-clinical; protein expression; relating to nervous system; social; synaptic function; therapeutic target; tomography; touchscreen; transmission process

DESCRIPTION (provided by applicant): Our central hypothesis is that developmental delay syndromes including autism spectrum disorders lead to alterations in synaptic function in integrative brain regions that result in aberrant behavioral phenotypes. We will explore this hypothesis in a genetically modified rat model. Haploinsufficiency of SHANK3 leads to neurodevelopmental changes that include autism spectrum disorders, attentional disorders, absent or delayed speech, mild to moderate intellectual disability, and motor alterations. The SHANK3 protein forms a key structural part of the postsynaptic density. Because of the closer physiology between rats and humans as compared to mice, rats remain the primary choice of the pharmaceutical industry for studying pharmacokinetic (PK) properties of novel drugs. In addition, rats provide a far more tractable experimental model system for neurobiological, electrophysiological and behavioral studies, and it is of course advantageous, when considering drug development, that the biological assays be done in the same species where the PK studies are carried out. We have used zinc-finger nucleases to develop a genetically engineered rat with a disruption in the full-length rat Shank3 gene. This represents a first-ever genetically modified rat model for ASD and permits us to carry out detail studies in the prefrontal cortex, an area of great importance in autism, not easily studied in mouse models. We propose to carry out a detailed analysis of this model. We plan to test our central hypothesis with the following specific aims: 1) Behavioral assessment of prefrontal function in Shank3-deficient rats; 2) Electrophysiological analysis of prefrontal function in Shank3-deficient rats; and, 3) Neuropathological and neurochemical investigation of prefrontal function in Shank3-deficient rats. 3) The research is innovative, in our opinion, because it will make use of a first-ever rat model of ASD. In addition, it is innovative in the use of state-of-the art approaches to understanding the role of PFC in ASD, a key region not yet studied in detail in ASD model systems. The focus on PFC also allows for studying neuronal pathways that feed into the PFC, including the first-ever behavioral neurophysiological assessment of hippocampal-prefrontal circuitry in a rodent model for ASD. Our approach to high-resolution 3D imaging and analysis of neuronal morphology down to the level of single spine is notably novel. This form of analysis will allow us to identify molecular targets that are affected in Shank3-deficient rats, in particular, te distribution of excitatory receptors and synaptic proteins known to be linked to spine and synapse size and maturity. Finally, our behavioral analyses will make use of novel touchscreen chambers for detailed analysis of PFC function.

描述（由申请人提供）：我们的中心假设是，包括自闭症谱系障碍在内的发育迟缓综合征会导致大脑整合区域突触功能的改变，从而导致异常的行为表型。我们将在转基因大鼠模型中探索这一假设。 SHANK3 的单倍体不足会导致神经发育变化，包括自闭症谱系障碍、注意力障碍、言语缺失或延迟、轻度至中度智力障碍和运动改变。 SHANK3 蛋白形成突触后密度的关键结构部分。由于与小鼠相比，大鼠和人类之间的生理学更为接近，因此大鼠仍然是制药行业研究新药药代动力学 (PK) 特性的主要选择。此外，大鼠为神经生物学、电生理学和行为研究提供了一个更容易处理的实验模型系统，在考虑药物开发时，在进行 PK 研究的同一物种中进行生物测定当然是有利的。我们使用锌指核酸酶培育了一种基因工程大鼠，其全长大鼠 Shank3 基因被破坏。这是第一个针对自闭症谱系障碍的转基因大鼠模型，使我们能够对前额皮质进行详细研究，前额皮质是自闭症的一个非常重要的区域，在小鼠模型中不容易研究。我们建议对该模型进行详细分析。我们计划通过以下具体目标来检验我们的中心假设：1）Shank3 缺陷大鼠前额叶功能的行为评估； 2）Shank3缺陷大鼠前额叶功能的电生理分析； 3) Shank3 缺陷大鼠前额叶功能的神经病理学和神经化学研究。 3）我们认为，这项研究具有创新性，因为它将利用第一个自闭症谱系障碍（ASD）大鼠模型。此外，它在使用最先进的方法来理解 PFC 在 ASD 中的作用方面具有创新性，这是 ASD 模型系统中尚未详细研究的关键区域。对 PFC 的关注还可以研究输入 PFC 的神经元通路，包括首次对 ASD 啮齿动物模型中的海马-前额叶回路进行行为神经生理学评估。我们对单个脊柱水平的神经元形态进行高分辨率 3D 成像和分析的方法非常新颖。这种形式的分析将使我们能够识别 Shank3 缺陷大鼠中受影响的分子靶标，特别是已知与脊柱和突触大小和成熟度相关的兴奋性受体和突触蛋白的分布。最后，我们的行为分析将利用新型触摸屏室对 PFC 功能进行详细分析。