Coupling neuroimaging with CLARITY and single cell genomics to dissect sex differences in the developing brain

将神经影像与 CLARITY 和单细胞基因组学结合起来，剖析大脑发育中的性别差异

• 批准号: 10360528
• 负责人: Arthur P Arnold
• 金额: $ 48.52万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10360528
• 关键词: Acrylamides; Address; Adolescent; Affect; Age; Anatomy; Animal Model; Area; Attention; Automobile Driving; Autopsy; Biological; Biology; Brain; Brain imaging; Brain region; Cell Size; Cells; Collaborations; Coupling; Data Set; Development; Disease; Dissection; Eating Disorders; Etiology; Female; Foundations; Genes; Genetic Models; Genomics; Genotype; Gonadal Hormones; Gonadal Steroid Hormones; Gonadal structure; Health; Homologous Gene; Hormonal; Hormones; Human; Hybrids; Hydrogels; Image; Impairment; Individual; Investigation; Kallmann Syndrome; Knowledge; Language; Language Development; Life; Lipids; Maps; Measures; Methods; Modeling; Molecular; Mood Disorders; Motor; Mus; Neurobiology; Outcome; Pathologic; Pathway interactions; Patients; Pattern; Ploidies; Population; Resolution; Risk; Role; Scanning; Sex Bias; Sex Chromosomes; Sex Differences; Sex Differentiation; Sex Factors; Signal Transduction; Site; Social Development; Social Functioning; Stereotyping; Testing; Time; Tissue Sample; Tissues; To specify; Translating; Trisomy; United States National Institutes of Health; Ursidae Family; Variant; Work; Y Chromosome; Yang; age related; analysis pipeline; brain magnetic resonance imaging; cell type; cohort; computerized tools; density; dosage; high throughput analysis; human model; in vivo; innovation; interdisciplinary collaboration; longitudinal analysis; male; motor control; mouse model; nervous system disorder; neurobehavioral; neurodevelopment; neuroimaging; novel; protective effect; sex; sex chromosome aneuploidy; single cell sequencing; single-cell RNA sequencing; spatiotemporal

Abstract Many neurobehavioral diseases affect females and males differently, and emerge at different stages of life, leading to the conclusion that one sex is protected from diseases by inherent sex factors, such as gonadal hormones and sex chromosomes. The sites and timing of these effects on brain development are unknown. The method of high-throughput high-precision whole-brain MRI imaging has the power to detect such changes with great sensitivity, in both animal models and humans. This project will exploit these powerful methods to separate gonadal hormonal and sex chromosome effects in the mouse model “Sex Chromosome Trisomy” (SCT), at 9 different ages of development, to discover where and when these factors cause sex differences in brain development (Aim 1). The SCT model compares mice with different numbers and types of sex chromosomes (XX, XY, XXY, XYY), each genotype present in gonadal males or females. Then, a novel pipeline of analysis will compare mouse and human brain development at many stages and in informative groups (differing by age, sex, hormonal status, and sex chromosome complement) to determine which changes in mouse brain are also found in humans, in specific brain regions related to different diseases (Aim 2). The analysis will point to changes in mouse brain that model human brain development. These studies will also pinpoint new sex-biasing effects of hormones and sex chromosomes at localized brain regions at specific developmental stages in mice, leading to further investigation of cellular and molecular changes caused by sex at those sites (Aim 3). Cellular effects (cell size, number, density of defined cell populations) of sex-biasing factors will be measured using CLARITY (Clear Lipid-exchanged Acrylamide-hybridized Rigid Imaging-compatible Tissue- hYdrogel). Gene pathways responding to hormonal and sex chromosome effects in individual cell types in specific brain regions will be measured using single cell RNA-seq. These studies, combining high- resolution neuroimaging, CLARITY, and single cell sequencing, will provide a foundation of concepts about where in the brain, and when during development, specific cell populations respond to sex factors, as a prelude for hypothesizing which sex differences underlie the protective effects of sex- biased factors in cells.

抽象的 许多神经行为疾病对女性和男性的影响不同，出现的时间也不同。 生命的各个阶段，得出这样的结论：固有的性别因素可以保护一种性别免受疾病的影响， 例如性腺激素和性染色体。这些对大脑影响的部位和时间 发展未知。高通量高精度全脑MRI成像方法 在动物模型和人类中以极高的灵敏度检测此类变化的能力。这 该项目将利用这些强大的方法来分离性腺激素和性染色体的影响 在小鼠模型“性染色体三体”（SCT）中，在 9 个不同的发育年龄，发现 这些因素在何时何地导致大脑发育存在性别差异（目标 1）。 SCT模型 比较具有不同数量和类型性染色体（XX、XY、XXY、XYY）的小鼠，每种基因型 存在于男性或女性的性腺中。然后，一种新颖的分析流程将比较小鼠和人类 大脑发育的多个阶段和信息群体（因年龄、性别、荷尔蒙状态和 性染色体补体）来确定小鼠大脑中的哪些变化也存在于人类中 与不同疾病相关的特定大脑区域（目标 2）。分析将指出鼠标的变化 模拟人类大脑发育的大脑。这些研究还将查明新的性别偏见影响 小鼠特定发育阶段局部大脑区域的激素和性染色体， 从而进一步研究这些部位由性别引起的细胞和分子变化（目标 3）。 性别偏见因素的细胞效应（细胞大小、数量、特定细胞群的密度）将是 使用 CLARITY（透明脂质交换丙烯酰胺杂交刚性成像兼容组织）进行测量 水凝胶）。个体细胞类型中响应激素和性染色体效应的基因途径 将使用单细胞 RNA-seq 对特定大脑区域进行测量。这些研究结合了高 分辨率神经影像、CLARITY 和单细胞测序将为概念提供基础 关于特定细胞群在大脑中的位置以及发育过程中的何时对性做出反应 因素，作为假设哪些性别差异是性别保护作用的前奏 细胞中的偏见因素。