Defining the genetic and cellular basis of morphological diversity in Drosophila

定义果蝇形态多样性的遗传和细胞基础

• 批准号: 10000206
• 负责人: Benjamin James Vincent
• 金额: $ 6.74万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-01 至 2022-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10000206
• 关键词: 3-Dimensional; Address; Adhesives; Affect; Animals; Behavior; Case Study; Cell Count; Cell Shape; Cell Size; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; DNA; Development; Developmental Gene; Diagnostic; Drosophila genus; Drosophila melanogaster; Evolution; Exhibits; Foundations; Gene Expression; Genes; Genetic; Genetic Complementation Test; Genetic Variation; Genomic Segment; Goals; Human; Hybrids; Image; Individual; Label; Learning; Limb structure; Link; Lobe; Male Genital Organs; Maps; Measures; Molecular; Morphogenesis; Morphology; Outcome; Pattern; Phenotype; Pigmentation physiologic function; Reagent; Regulation; Regulator Genes; Research; Resolution; Shapes; Signal Transduction; Signaling Molecule; Structure; System; Tissues; Variant; Work; base; causal variant; comparative; expectation; experimental study; fly; genetic variant; genome editing; human disease; mutant; three dimensional structure; trait; transcription factor; transcriptome sequencing; transcriptomics

PROJECT SUMMARY The overall goal of this proposal is to determine the genetic variants that alter complex three-dimensional morphologies and to determine how these genes exert their influence on cell number, shape and behavior during development. Developmental genes that are important in morphogenesis are linked in complex hierarchies, yet we do not currently know how to prioritize variants within the context of a gene regulatory network. I will therefore undertake an evolutionary case study at the level of a gene regulatory network to determine the types of genes that contribute to morphological diversification between species and characterize their effects on individual cells. To accomplish this goal, I require a system where the gene regulatory network underlying a quantitative trait is well-understood, and where critical variants can be mapped at the resolution of individual genes. The posterior lobe in the Drosophila melanogaster clade is particularly well-suited to connecting genetic variation to morphogenetic outcomes. Recent work has identified many components of the gene regulatory network controlling posterior lobe development, including patterning molecules (transcription factors and signaling molecules) and cellular effectors (cytoskeletal regulators). Furthermore, hybrids between all three species are viable and exhibit intermediate lobe morphologies, which allows for rigorous mapping of genomic regions contributing to their drastically different phenotypes. In Aim 1, I will map sequences responsible for quantitative variation in posterior lobe morphology. I will use RNA-seq in species and hybrids to identify genes containing expression variation, with the expectation that some of these genes contribute to morphological diversity in the posterior lobe. I will then validate strong candidates using a CRISPR-based complementation test. In Aim 2, I will determine how patterning molecules and cellular effectors influence the number, shape and behavior of individual cells. I will use cell labeling and imaging to measure cell number, size and behavior in posterior lobes derived from mutants, different species and species hybrids. By completing this proposal, I will connect variation in the gene regulatory network to molecular mechanisms that dictate cell size and shape, which will develop the posterior lobe as a premier system for systems-level studies of morphological development and evolution.

项目摘要 该提案的总体目标是确定改变复杂三维的遗传变异 形态学并确定这些基因如何对细胞数，形状和行为产生影响 在开发过程中。在形态发生中很重要的发育基因在复合物中有联系 层次结构，但是我们目前不知道如何在基因调节的背景下优先考虑变体 网络。因此，我将在基因调节网络级别进行进化案例研究 确定有助于物种形态多样化的基因类型并表征 它们对单个细胞的影响。为了实现这一目标，我需要一个基因调节网络的系统 基本的定量性状是众所周知的，可以将关键变体映射到分辨率上 单个基因。 Melanogaster进化果中的后叶特别适合 将遗传变异与形态发生结局联系起来。最近的工作已经确定了许多组成部分 基因调节网络控制后叶发育，包括图案分子（转录 因素和信号分子）和细胞效应子（细胞骨架调节剂）。此外，之间的杂种 这三个物种都是可行的，并且表现出中间叶的形态，可以进行严格的映射 基因组区域有助于其截然不同的表型。在AIM 1中，我将映射序列 负责后叶形态的定量变化。我将在物种中使用RNA-seq， 鉴定包含表达变异的基因的杂种，并期望其中一些基因 有助于后叶的形态多样性。然后，我将使用一个 基于CRISPR的互补测试。在AIM 2中，我将确定如何进行分子和细胞的模式 效应子影响单个细胞的数量，形状和行为。我将使用细胞标签和成像 测量源自突变体，不同物种和的后叶的细胞数，大小和行为 物种杂种。通过完成此建议，我将将基因调节网络中的变化连接到 决定细胞大小和形状的分子机制，这将发展为主要叶 系统级研究形态发展和进化的系统。