In toto Image Analysis of Tissue Mechanics during Vertebrate Ear Development

脊椎动物耳朵发育过程中组织力学的整体图像分析

• 批准号: 8442611
• 负责人: Kishore Rao Mosaliganti
• 金额: $ 9.9万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-06-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8442611
• 关键词: Address; Adhesions; Adopted; Affect; Automobile Driving; Award; Behavior; Biochemical Process; Biological Models; Cadherins; Cell Adhesion; Cell Adhesion Molecules; Cell Count; Cell Polarity; Cell Shape; Cell Volumes; Cell division; Cell model; Cell physiology; Cell-Cell Adhesion; Cells; Cellular Morphology; Clinical; Computer Simulation; Cytoskeleton; DNA Sequence Rearrangement; Data; Development; Dose; Ear; Early treatment; Ectoderm; Elements; Endolymph; Epithelial; Equilibrium; Etiology; Four-dimensional; Furosemide; Generations; Genes; Genetic; Genetic Models; Glass; Goals; Growth; Hearing; Image; Image Analysis; Imaging technology; Individual; Injection of therapeutic agent; Ion Transport; Ions; Labyrinth; Lateral; Learning; Liquid substance; Location; Measures; Mechanics; Microscope; Microscopy; Modeling; Molecular; Monitor; Morphogenesis; Morphology; Na(+)-K(+)-Exchanging ATPase; Nuclear Envelope; Onions; Organ; Organizational Change; Osmotic Pressure; Otic Placodes; Otic Vesicle; Outcome; Pattern; Pharmacodynamics; Play; Process; Recruitment Activity; Regulation; Reporter; Research; Resolution; Role; Semicircular canal structure; Shapes; Staging; Structure; Surface Tension; System; Technology; Testing; Thick; Time; Tissues; Transgenic Organisms; Vertebrates; Vesicle; Work; Zebrafish; base; deafness; hindbrain; insight; interest; ion channel blocker; migration; movie; multi-scale modeling; mutant; otoconia; pressure; protein expression; public health relevance; skills; spatiotemporal; subcutaneous

DESCRIPTION (provided by candidate): In toto Image Analysis of Tissue Mechanics during Vertebrate Ear Morphogenesis Understanding ear development is key to developing clinical therapies for early/late-onset deafness, whether of genetic or environmental etiology. The goal of the proposed study is to determine the mechanical contribution of patterning circuits involved in vertebrate ear development. My focus is on the morphogenesis of the otic-vesicle tissue, which plays a fundamental role in the generation of specialized organs responsible for the sense of hearing and balance. The otic vesicle develops from an initially thickened ectodermal placode that is induced to form a subcutaneous, hollow epithelialized shell filled with endolymph. The proposed research uses the zebrafish model system and addresses three fundamental questions: (A) How does the otic tissue transform from a sheet of newly recruited otic placode cells into radially polarized epithelial tissue with a specific tissue/cell morphology (B) What is the role of endolymph pressure in driving the growth of the otic vesicle to the right shape/size? As such, large- scale organizational changes arising from changes in cell number and localized cell shape are observable along the otic-vesicle perimeter. (C) How does endolymph pressure equilibrate with intercellular forces of adhesion and cortical tension that affect cell/tissue shape? To address these questions, the following three aims are proposed: (1) I propose to conduct in toto imaging with fluorescent reporters to generate 4-dimensional cellular descriptions of the morphogenesis process. By using image analysis, I will comprehensively reconstruct the locations, tracks, and cell divisions of placode cells involved in otic-vesicle formation and learn how otic placode cells delaminate from ectoderm tissue and rearrange into radially polarized hollow tissue. (2) I will model the otic vesicle as a "pressurize shell" using a finite-element representation to understand tissue mechanics of otic-vesicle growth. A finite-element cell model will reveal how rearrangement forces from endolymph secretion drive tissue shape change. (3) I will integrate the model with the function of patterning genes for transepithelial transport and inter- cellular adhesion. By using the biophysical model, I will test model outcomes with pharmacological and genetic perturbations (mutants+morphants) by altering cell-cell adhesion protein expression and endolymph pressure. These perturbations will demonstrate how patterning circuits at a cellular level control specific aspects of the morphogenesis process. Together, these aims will elucidate how patterning inputs coordinate cell mechanics for ear morphogenesis by controlling endolymph secretion and spatiotemporal adhesion protein expression.

描述（由候选人提供）：在脊椎动物耳朵形态发生期间的组织力学图像分析中，理解耳朵发育是开发早期/晚发耳聋的临床疗法的关键，无论是遗传性或环境病因。拟议的研究的目的是确定参与脊椎动物耳朵发育的模式电路的机械贡献。我的重点是针对耳虫组织的形态发生，它在负责听力和平衡感的专门器官的产生中起着基本作用。耳囊是由最初增厚的外胚层斑点形成的，该外胚层被诱导形成皮下，空心的上皮壳，充满了内膜。拟议的研究使用斑马鱼模型系统，并解决了三个基本问题：（a）眼膜组织如何从新募集的OTIC OTECODE PLACODE细胞转变为具有特定的组织/细胞形态的径向极化上皮组织（B）内接晶压力在驱动Otot Veesicle的生长中的作用是什么？因此，可观察到细胞数和局部细胞形状变化的大规模组织变化沿着眼膜周长可观察到。 （c）内淋巴压力如何与影响细胞/组织形状的粘附和皮质张力的细胞间力平衡？为了解决这些问题，提出了以下三个目的：（1）我建议用荧光记者进行Toto成像进行成像，以生成形态发生过程的4维细胞描述。通过使用图像分析，我将全面地重建参与耳虫形成的位置细胞的位置，轨道和细胞划分，并了解耳片斑块细胞如何从外胚层组织分配并重新排列到径向极化的空心组织。 （2）我将使用有限元代表形式将耳囊囊泡作为“加压壳”建模，以了解耳囊生长的组织力学。有限元细胞模型将揭示内淋巴分泌的重排力如何驱动组织形状变化。 （3）我将将模型与图案的功能集成 旋转转运和细胞间粘附的基因。通过使用生物物理模型，i 将通过改变细胞 - 细胞粘附蛋白表达和内淋巴压力来测试用药理学和遗传扰动（突变体+形态）的模型结果。这些扰动将证明在细胞水平上模仿电路如何控制形态发生过程的特定方面。总之，这些目标将通过控制内淋巴的分泌和时空粘附蛋白表达来阐明模式输入如何将细胞力学配置为耳朵形态发生。