权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Morphogenesis: Biophysics and Genetics of Dorsal Closure

形态发生：背侧闭合的生物物理学和遗传学

基本信息

批准号：
10623612
负责人：
DANIEL PETER KIEHART
金额：
$ 51.74万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-07-01 至 2028-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10623612
关键词：
Actomyosin Address Adhesions Animals Biochemical Biological Biological Models Biological Process Biophysical Process Biophysics Cell Communication Cell Shape Cells Cellular biology Characteristics Chromosomes Cleft Palate Complex Cytoskeleton Defect Development Developmental Process Dorsal Drosophila genus Drosophila melanogaster Embryo Epithelium Functional disorder Gene Expression Genes Genetic Genome Goals Heart Homeostasis Human Pathology Image Instruction Kinetics Lateral Lesion Mechanics Molecular Molecular Probes Morphogenesis Motor Movement Mutation Myosin ATPase Myosin S-2 Neural tube Palate Phenotype Phylogeny Positioning Attribute Predisposition Process Proteins RNA Splicing Regulation Research Shapes Signal Transduction Spinal Dysraphism Sum Thermodynamics Tissues Work biophysical techniques design fly forward genetics gastrulation gene product imaging genetics insight interdisciplinary approach interest mutant resilience reverse genetics stomach cardia transmission process wound wound healing

项目摘要

Cell sheet morphogenesis is essential for metazoan development and homeostasis of animal form – it contributes to development, such as in gastrulation, neural tube, heart and palate formation and to homeostasis, in wound healing. Gene expression and signaling cascades coordinate and regulate the cellular machines that drive morphogenesis. Disfunction in these components causes developmental and wound healing defects that can disfigure or kill. We focus on the molecular mechanisms of cell sheet morphogenesis during dorsal closure (DC) in Drosophila melanogaster. During DC, lateral epidermal sheets advance to close a dorsal opening. We pioneered the study of DC as a model system and use an unusually diverse repertoire of interdisciplinary approaches, including live imaging, reverse and forward genetics and biophysical strategies to interrogate the mechanics and regulation of closure in wild type and mutant embryos. We found that DC is the sum of four major dynamic processes and is robust and resilient – no single force that drives closure is absolutely required. Processes that contribute to DC at the molecular, cellular and tissue scales are highly conserved in animal phylogeny making Drosophila an ideal model system for interrogating the molecular basis of morphogenesis. There remain significant gaps in our understanding of this conceptually simple, yet biologically complex cell sheet movement. To identify new “DC genes”, i.e., genes that when deleted, disrupt closure, we initiated a forward genetic, live-imaging, screen. This screen used 194 deficiency stocks (Dfs) that collectively delete 5,778 of the 5,854 genes on melanogaster's 2nd chromosome. We have begun to extend our screen to the 3rd chromosome. Remarkably, 96 Dfs caused notable and diverse defects in closure, indicating that a large number of discrete biological processes contribute to closure and are susceptible to mutational disruption. Thus far, we have identified 13 new pre-DC or DC genes that are responsible for the DC Df phenotypes. When extended to the whole fly genome our screen is projected to identify ~165 new DC genes (only ~140 DC genes were known at the start of our screen). Based on phenotype, we prioritize the DC Dfs on which to focus, identify the DC genes responsible for their Df phenotypes, then characterize how the new DC gene products contribute to closure. A priority is to understand the molecular mechanisms by which cell-cell interactions and cell-matrix based adhesion couple to the actomyosin cytoskeleton – these connections must be robust enough to transmit forces, yet malleable enough to allow the cell shape changes that define morphogenesis. Of further interest is a new effort to thermodynamically and kinetically characterize the myosin 2 motor that drives morphogenesis in DC and other developmental processes. Our goal is to assess how differential splicing that encodes myosin's motor domain contributes to morphogenesis as a fast moving, slow/efficient force holding, strain sensing, or processive motor. We are uniquely positioned to address the molecular and biophysical mechanisms that underlie the basic biology of cell sheet morphogenesis in flies, research that directly informs vertebrate development and wound healing.

细胞片形态发生对于后生动物的发育和动物形态的稳态至关重要——它有助于发育，例如原肠胚形成、神经管、心脏和腭的形成以及伤口的稳态康复。基因表达和信号级联协调和调节驱动细胞的机器形态发生。这些成分的功能障碍会导致发育和伤口愈合缺陷毁容或杀死。我们重点研究背侧闭合（DC）过程中细胞片形态发生的分子机制在黑腹果蝇中。在 DC 期间，侧表皮片前进以关闭背侧开口。我们率先将 DC 作为模型系统进行研究，并使用了异常多样化的跨学科知识库方法，包括实时成像、反向和正向遗传学以及生物物理策略来询问野生型和突变体胚胎的闭合机制和调节。我们发现DC是四个主要的总和动态过程，坚固耐用，有弹性——绝对不需要单一的力量来驱动关闭。在分子、细胞和组织尺度上促进 DC 的过程在动物中高度保守系统发育使果蝇成为研究形态发生分子基础的理想模型系统。我们对这种概念上简单但生物学上复杂的细胞的理解仍然存在重大差距片材运动。为了识别新的“DC 基因”，即删除时会破坏闭合的基因，我们启动了一个前向研究遗传、实时成像、筛查。该屏幕使用了 194 个缺陷库存 (Dfs)，总共删除了 5,778 个黑腹果蝇第二条染色体上有 5,854 个基因。我们已经开始将筛选范围扩大到第三条染色体。值得注意的是，96 个 Dfs 在闭合中造成了显着且多样的缺陷，表明大量离散生物过程有助于闭合并且容易受到突变破坏。到目前为止，我们已经鉴定了 13 个新的前 DC 或 DC 基因，它们负责 DC Df 表型。当扩展到整体时果蝇基因组我们的屏幕预计将识别约 165 个新的 DC 基因（一开始只知道约 140 个 DC 基因）我们的屏幕）。根据表型，我们优先考虑要关注的 DC Dfs，确定负责的 DC 基因确定它们的 Df 表型，然后描述新的 DC 基因产物如何有助于闭合。首要任务是了解细胞间相互作用和基于细胞基质的粘附耦合的分子机制肌动球蛋白细胞骨架——这些连接必须足够坚固以传递力，但又具有足够的可塑性允许定义形态发生的细胞形状变化。更令人感兴趣的是一项新的努力热力学和动力学表征了驱动直流和其他形态发生的肌球蛋白 2 马达发展过程。我们的目标是评估差异剪接如何编码肌球蛋白的运动域作为快速移动、缓慢/高效的力保持、应变感应或处理马达，有助于形态发生。我们拥有独特的优势来解决基础生物学的分子和生物物理机制果蝇细胞片形态发生的研究，直接影响脊椎动物的发育和伤口愈合。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Identifying Genetic Players in Cell Sheet Morphogenesis Using a Drosophila Deficiency Screen for Genes on Chromosome 2R Involved in Dorsal Closure.

使用果蝇缺陷筛选 2R 号染色体上参与背侧闭合的基因来识别细胞片形态发生中的遗传因素。

DOI：
10.1534/g3.118.200233
发表时间：
2018
期刊：
G3 (Bethesda, Md.)
影响因子：
0
作者：
Mortensen,RichardD;Moore,ReganP;Fogerson,StephanieM;Chiou,HellenY;Obinero,ChimdinduV;Prabhu,NeelK;Wei,AngelaH;Crawford,JaniceM;Kiehart,DanielP
通讯作者：
Kiehart,DanielP

Superresolution microscopy reveals actomyosin dynamics in medioapical arrays.

DOI：
10.1091/mbc.e21-11-0537
发表时间：
2022-09-15
期刊：
MOLECULAR BIOLOGY OF THE CELL
影响因子：
3.3
作者：
Moore, Regan P.;Fogerson, Stephanie M.;Tulu, U. Serdar;Yu, Jason W.;Cox, Amanda H.;Sican, Melissa A.;Li, Dong;Legant, Wesley R.;Weigel, Aubrey, V;Crawford, Janice M.;Betzig, Eric;Kiehart, Daniel P.
通讯作者：
Kiehart, Daniel P.

Identifying Key Genetic Regions for Cell Sheet Morphogenesis on Chromosome 2L Using a Drosophila Deficiency Screen in Dorsal Closure.

DOI：
10.1534/g3.120.401386
发表时间：
2020-11-05
期刊：
G3 (Bethesda, Md.)
影响因子：
0
作者：
Fogerson SM;Mortensen RD;Moore RP;Chiou HY;Prabhu NK;Wei AH;Tsai D;Jadi O;Andoh-Baidoo K;Crawford J;Mudziviri M;Kiehart DP
通讯作者：
Kiehart DP

Mutations in Drosophila crinkled/Myosin VIIA disrupt denticle morphogenesis.

果蝇皱纹/肌球蛋白VIIA 的突变破坏了小齿的形态发生。

DOI：
10.1016/j.ydbio.2020.11.007
发表时间：
2021-03
期刊：
Developmental biology
影响因子：
2.7
作者：
Sallee JL;Crawford JM;Singh V;Kiehart DP
通讯作者：
Kiehart DP

Wound repair in sea urchin larvae involves pigment cells and blastocoelar cells.

海胆幼虫的伤口修复涉及色素细胞和胚泡细胞。

DOI：
10.1016/j.ydbio.2022.08.005
发表时间：
2022-11
期刊：
DEVELOPMENTAL BIOLOGY
影响因子：
2.7
作者：
Allen, Raymond L.;George, Andrew N.;Miranda, Esther;Phillips, Taji M.;Crawford, Janice M.;Kiehart, Daniel P.;McClay, David R.
通讯作者：
McClay, David R.