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.

项目总结