Development and function of Crhbp+ neurons in the zebrafish hypothalamus

斑马鱼下丘脑 Crhbp 神经元的发育和功能

• 批准号: 10611936
• 负责人: Priscilla Figueroa
• 金额: $ 3.82万
• 依托单位: UNIVERSITY OF UTAH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-05-01 至 2024-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10611936
• 关键词: Ablation; Affect; Anxiety; Area; Behavior; Behavioral; Biological Assay; Biological Models; Brain; C-terminal; CRISPR/Cas technology; Cell Transplantation; Cells; Chimera organism; Cognition; Communication; Darkness; Data; Development; Disease; Diving; Embryo; Endocrine; Enzyme-Linked Immunosorbent Assay; Exhibits; Exons; Exploratory Behavior; Frameshift Mutation; Future; Gastrula; Gene Expression Regulation; Generations; Genes; Genetic; Genetic Transcription; Goals; Homeostasis; Hormones; Human; Hydrocortisone; Hypothalamic structure; Individual; Investigation; Knock-out; Larva; Link; Measures; Mediating; Mediator; Mental Depression; Mentors; Messenger RNA; Metronidazole; Molecular; Mood Disorders; Moods; Mus; Mutation; Names; Neurons; Nitroreductases; Pathway interactions; Peripheral; Phenotype; Physiology; Pituitary Gland; Play; Population; Posterior Hypothalamus; Procedures; Protein Isoforms; RNA Splicing; Recovery; Regulation; Reporter; Research; Research Personnel; Research Proposals; Role; Scientist; Series; Signal Pathway; Stress; Tail; Testing; Tissues; Training; Transgenes; Transgenic Organisms; Transplantation; Vertebrates; Work; Zebrafish; beta catenin; biological adaptation to stress; blastomere structure; career; corticotropin releasing factor-binding protein; embryo cell; experimental study; mutant; neurogenesis; neuron development; neuronal circuitry; neuropsychiatric disorder; novel; overexpression; response; skills; student mentoring; training opportunity; transcription factor; transcriptome sequencing

Project Summary/Abstract The vertebrate hypothalamus maintains physiological homeostasis through regulation of the stress response pathway. Disruption of this pathway can lead to neuropsychiatric disorders, but many of the genes and neurons involved are unidentified. Our lab has discovered that the Wnt pathway mediator Lef1 regulates hypothalamic neurogenesis and stress-related exploratory behavior, and my preliminary data suggest that a specific Lef1 protein isoform enriched in the developing brain may play a unique role in both of these functions. The development of hypothalamic neurons expressing corticotropin-releasing hormone binding protein (Crhbp) depends on Lef1 as well as its transcriptional target otbp, providing a hypothesized mechanism linking Wnt- dependent transcription to behavior. This research proposal will test this hypothesis while providing technical and career training essential for my goal to become an independent academic researcher. In my first Aim I will create mutations disrupting specific Lef1 isoforms, and assess their effects on hypothalamic neurogenesis, physiology, and behavior. In my second Aim, I will use cell transplantation to create genetic chimeras, and determine whether Crhbp+ cells are necessary and sufficient to regulate exploratory behavior downstream of Lef1 and Otpb. From this project I will identify molecules and circuits that may be conserved between zebrafish and human stress response pathways, and gain new skills that will allow me to become a successful scientist and mentor.

项目摘要/摘要 脊椎动物下丘脑通过调节压力反应维持生理稳态 路径。这种途径的破坏会导致神经精神疾病，但许多基因和神经元 涉及的不明身份。我们的实验室发现WNT途径介质LEF1调节下丘脑 神经发生和与压力相关的探索行为，我的初步数据表明特定的LEF1 富含发展大脑的蛋白质同工型在这两个功能中都可能发挥独特的作用。这 表达皮质激素释放激素结合蛋白（CRHBP）的下丘脑神经元的发展 取决于LEF1及其转录靶标OTBP，提供了与Wnt-相关的假设机制 依赖转录对行为。该研究建议将在提供技术的同时检验该假设 职业培训对于我的目标是成为一名独立学术研究员的目标。在我的第一个目标中，我会 创建破坏特定LEF1同工型的突变，并评估其对下丘脑神经发生的影响， 生理和行为。在我的第二个目标中，我将使用细胞移植来创建遗传嵌合体，并 确定CRHBP+细胞是否必要且足以调节下游的探索性行为 LEF1和OTPB。从这个项目中，我将确定斑马鱼之间可能保守的分子和电路 人力压力反应途径，并获得新技能，使我成为一名成功的科学家 和导师。