Genetic and functional analysis of eye degeneration in natural populations of a cave-dwelling crustacean

洞穴甲壳动物自然种群眼睛退化的遗传和功能分析

基本信息

批准号：
9812713
负责人：
Meredith E Protas
金额：
$ 33.28万
依托单位：
DOMINICAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-01 至 2023-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9812713
关键词：
Affect Albinism Alleles Animals Anophthalmos Biological Models Birth Candidate Disease Gene Clustered Regularly Interspaced Short Palindromic Repeats Congenital Disorders Crustacea DNA Sequence Alteration Data Development Developmental Biology Disease Embryo Evolution Eye Eye Development Genes Genetic Human Hybrids Invertebrates Isopoda Lead Methods Microphthalmos Modeling Molecular Morphology Mus Mutagenesis Mutate Mutation Natural regeneration Pathway interactions Patients Phenotype Pigments Play Population Process Reagent Research Role Sampling Study models Surface Techniques Testing Time Variant Visual system structure Work base comparative developmental disease developmental genetics differential expression eye formation fly functional loss gene discovery genome editing human disease human model mutant novel novel strategies tool transcriptome transcriptome sequencing

项目摘要

Abstract: An understanding of eye development is necessary to comprehend proper functioning of the eye, capacity for regeneration, and the developmental basis of disease. Many model systems, both vertebrate and invertebrate, have historically provided much important data about the developmental and genetic basis of eye formation. However, hurdles exist studying eye development in model systems such as inherent biases in mutant screens and the minimal amount of naturally occurring variation in eye size. One novel approach allowing for a greater understanding of the developmental and genetic bases of eye formation is to examine naturally occurring eye size variation in non-model organisms, such as that occurs in cave animals. Our previous work demonstrated the utility of using non-model organisms to model human disease when we found that a commonly mutated gene in cases of human albinism was also responsible for albinism in the cavefish, Astyanax mexicanus. We initially chose the isopod crustacean, Asellus aquaticus as a model for studying eye variation because of the extreme difference in eye size between cave and surface dwelling populations. Our subsequent work on the species demonstrated further advantages including multiple independent mechanisms of eye reduction/ loss within a single cave population, multiple mechanisms of eye loss between different cave populations, and the large genetic component influencing eye size. We have already generated many molecular, developmental, and genetic tools and reagents that will assist in studying this interesting species. Our proposal aims to understand the genetics behind eye degeneration in this species. First, we use comparative transcriptome sequencing of cave and surface embryonic samples as an unbiased method to identify genes and pathways responsible for eye degeneration in the cave form. Second, we sequence transcriptomes of hybrid embryos to identify genes that show allele specific expression and that putatively contain cis-regulatory changes. Next, we validate genes discovered from the transcriptome sequencing using linkage and functional techniques. Our work provides a novel perspective on the developmental and genetic basis of eye size differences and furthers the development of A. aquaticus as an important model for eye degeneration and disease.

抽象的：必须了解眼睛发育，以理解适当的功能眼睛，再生能力和疾病的发育基础。许多模型脊椎动物和无脊椎动物的系统历史上都提供了非常重要的有关眼睛形成的发育和遗传基础的数据。但是，障碍存在研究模型系统中的眼睛发育，例如突变体的固有偏见屏幕和眼睛尺寸自然发生的自然变化量最小。一本小说方法允许对发展和遗传基础有更深入的了解眼睛形成是检查非模型中自然发生的眼睛尺寸变化有机体，例如在洞穴动物中发生的生物。我们以前的工作证明了当我们发现一个在人白化病的病例中通常是突变的基因也是白化病的原因在山洞鱼中，阿斯蒂亚纳克斯墨西哥。我们最初选择了Isopod甲壳类动物Asellus aquaticus作为研究眼睛的模型由于洞穴和表面之间的眼睛尺寸极大差异而变化住宅人群。我们随后在该物种的工作进一步证明优点，包括多种独立的眼睛减少/损失机制单个洞穴种群，不同洞穴之间的多种眼睛流失机制种群以及影响眼睛大小的大遗传成分。我们已经生成了许多分子，发育和遗传工具以及试剂协助研究这个有趣的物种。我们的建议旨在了解遗传学该物种的眼睛变性后面。首先，我们使用比较转录组洞穴和表面胚胎样品的测序作为一种无偏的方法来识别导致洞穴形式的眼睛变性的基因和途径。第二，我们混合胚胎的序列转录组，以鉴定显示出等位基因特异性的基因表达和推理的包含顺式调节的变化。接下来，我们验证基因使用链接和功能技术从转录组测序中发现。我们的工作为眼睛的发育和遗传基础提供了一种新颖的观点大小差异，并进一步发展水曲霉作为重要模型眼睛变性和疾病。