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.

项目总结/摘要 脊椎动物下丘脑通过调节应激反应来维持生理稳态 通路这条通路的中断会导致神经精神疾病，但许多基因和神经元 涉案人员身份不明。本实验室发现Wnt通路介导因子Lef 1调节下丘脑 神经发生和压力相关的探索行为，我的初步数据表明，一个特定的Lef 1 在发育中的脑中富集的蛋白质同种型可能在这两种功能中发挥独特的作用。的 表达促肾上腺皮质激素释放激素结合蛋白（Crhbp）的下丘脑神经元的发育 依赖于Lef 1及其转录靶点otbp，提供了一种假设的连接Wnt- 行为的依赖性转录。这项研究计划将测试这一假设，同时提供技术支持。 和职业培训对我成为一名独立的学术研究者的目标至关重要。在我的第一个目标，我将 创建破坏特定Lef 1亚型的突变，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造遗传嵌合体， 确定Crhbp+细胞是否是调节下游探索行为所必需和足够的。 Lef 1和Otpb。从这个项目中，我将确定分子和电路，可能是保守的斑马鱼之间 和人类的压力反应途径，并获得新的技能，使我成为一个成功的科学家 和导师