Model behavior in zebrafish: characterization of the startle response

斑马鱼的模型行为：惊吓反应的表征

基本信息

批准号：
10376471
负责人：
Joy Hart Meserve
金额：
$ 3.43万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-01-01 至 2021-08-16
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10376471
关键词：
Acetylcholine Acoustics Affect Alleles Anatomy Animal Model Attention Deficit Disorder Attention deficit hyperactivity disorder Auditory Behavior Behavior Disorders Behavioral Behavioral Symptoms Biological Assay Birth Brain CRISPR/Cas technology Cells Characteristics Choline Clinical Complex Defect Development Disease Family Fire - disasters Foundations Functional disorder Genes Genetic Genetic Models Genetic Screening Head Human Image Individual Laboratories Larva Lesion Major Depressive Disorder Membrane Transport Proteins Mental disorders Modeling Molecular Molecular Cloning Motor Motor Neurons Movement Mus Muscle Mutation Neurons Neurophysiology - biologic function Neurotransmitters Pathway interactions Phenotype Population Positioning Attribute Posture Presynaptic Terminals Production Reaction Reaction Time Regulation Response Latencies Response to stimulus physiology Role Schizophrenia Skeletal Muscle Speed Spinal Spinal Cord Startle Reaction Stimulus System Tactile Tail Techniques Tissues Transgenic Organisms Vertebrates Zebrafish auditory stimulus autism spectrum disorder behavioral phenotyping behavioral plasticity behavioral study choline transporter cholinergic experimental study gene function genome sequencing habituation hindbrain human disease in vivo insight member mutant nervous system disorder neural circuit neurodevelopment neuronal circuitry neuropsychiatric disorder programs promoter response solute stimulus sensitivity sustained attention whole genome

项目摘要

ABSTRACT Although behavioral deficits are common in neurological disorders, the genetic pathways and neural circuits underlying behavior are largely unknown. A behavior that is disrupted in numerous disorders including attention deficit disorder, autism spectrum disorders, and schizophrenia is the startle response. Following an intense auditory stimulus, a short latency response occurs wherein rapid muscle activation produces a defensive posture. Disrupted startle responses likely result from a broad defect in genes or circuits controlling behavior. The genetic contribution to behavioral symptoms in humans has been challenging to uncover, in part because they result from multigenic disorders affecting the vastly complex human brain. Genetically tractable animal models have emerged as a valuable system for investigating gene function in the development and function of neural circuits underlying behavior. The Granato lab performed a genetic screen for regulators of the startle response in the larval zebrafish. Molecular cloning of 15 screen candidates identified a number of genes that modulate the startle response in zebrafish and are implicated in human disease. One such gene is solute carrier family 5 member 7a (slc5a7a), which encodes high affinity choline transporter that is required for synthesis of the neurotransmitter acetylcholine. Mutations in slc5a7 have been implicated in psychiatric disorders including attention deficit disorder and major depression. Mice lacking slc5a7 die immediately after birth, precluding behavioral studies. This proposal aims to determine the contribution of slc5a7a to neural circuit development and/or execution of the zebrafish startle response (Aims 1 and 2). In wild type larval zebrafish, the startle response begins with a sharp turn that initiates at the head and progresses towards the tail. In slc5a7a mutants, the turn initiates randomly along the body axis, resulting in an uncoordinated movement. This phenotype suggests neural circuitry between the hindbrain command neuron that initiates the movement and the downstream spinal motoneurons may be disrupted. Circuit defects in slc5a7a mutants will be investigated using in vivo Ca2+ imaging from individual neurons known to be required for the startle response and whole brain activity mapping of additional required neurons. The developmental stage and anatomical region where slc5a7a is required will be investigated using a combination of transgenic rescue constructs and CRISPR/Cas9 generated mutant alleles. Finally, a number of additional mutants that display uncoordinated or weak startle responses will be sequenced to determine the causative genetic lesion and will be characterized using the same techniques (Aim 3). The results of these proposed experiments will determine how slc5a7a promotes a coordinated startle response. More broadly, the results will elucidate acetylcholine’s role in regulating behavior. Finally, characterization of slca5a7a and genes identified in Aim 3 will contribute to models of the genetic program and neural circuitry underlying vertebrate startle behavior.

抽象的尽管行为缺陷在神经系统疾病中很常见，但遗传途径和神经系统疾病行为背后的电路在很大程度上是未知的，这种行为在许多疾病中被破坏，包括。注意力缺陷障碍、自闭症谱系障碍和精神分裂症是惊吓反应。强烈的听觉刺激，会发生短暂的潜伏反应，快速的肌肉激活产生防御性；惊吓反应中断可能是由控制行为的基因或电路的广泛缺陷造成的。基因对人类行为症状的影响一直难以揭示，部分原因是它们是由影响极其复杂的人类大脑的多基因疾病引起的。模型已成为研究基因在发育和功能中的功能的有价值的系统格拉纳托实验室对惊吓调节因子进行了基因筛查。 15 个候选斑马鱼的分子克隆鉴定出许多基因调节斑马鱼的惊吓反应并与人类疾病有关。家族 5 成员 7a (slc5a7a)，编码合成胆碱所需的高亲和力胆碱转运蛋白 slc5a7 的神经递质乙酰胆碱突变与精神疾病有关，包括缺乏 slc5a7 的小鼠会在出生后立即死亡，从而排除注意力缺陷障碍和重度抑郁症。该提案旨在确定 slc5a7a 对神经回路发育的贡献。和/或执行斑马鱼惊吓反应（目标 1 和 2）。在野生型斑马鱼幼体中，惊吓反应始于头部的急剧转动，在 slc5a7a 突变体中，转动沿着身体轴随机开始，从而产生一个这种表型表明后脑命令神经元之间存在神经回路。 slc5a7a 中启动运动的下游脊髓运动神经元可能会受到干扰。将使用已知的单个神经元的体内 Ca2+ 成像来研究突变体额外所需神经元的惊吓反应和全脑活动图。将使用转基因救援的组合来研究需要 slc5a7a 的解剖区域最后，一些额外的突变体被展示出来。将对不协调或微弱的惊吓反应进行测序，以确定致病病变的遗传因素，并将使用相同的技术进行表征（目标 3）。这些提议的实验的结果将确定。更广泛地说，这些结果将阐明 slc5a7a 如何促进协调的惊吓反应。最后，目标 3 中确定的 slca5a7a 和基因的表征将有助于脊椎动物惊吓行为的遗传程序和神经回路模型。