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）：脊椎动物耳朵形态发生过程中组织力学的整体图像分析了解耳朵发育是开发早发/晚发性耳聋临床治疗的关键，无论是遗传还是环境病因。拟议研究的目标是确定参与脊椎动物耳朵发育的图案电路的机械贡献。我的重点是耳泡组织的形态发生，它在负责听觉和平衡感的专门器官的生成中发挥着基础作用。耳囊由最初增厚的外胚层基板发展而来，经诱导形成皮下、充满内淋巴的中空上皮化外壳。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）耳组织如何从一片新招募的耳基板细胞转变为具有特定组织/细胞形态的径向极化上皮组织（B）什么是内淋巴压力在推动耳囊生长至正确形状/大小方面的作用？因此，沿着耳囊周边可以观察到由细胞数量和局部细胞形状的变化引起的大规模组织变化。 (C) 内淋巴压力如何与影响细胞/组织形状的细胞间粘附力和皮质张力平衡？为了解决这些问题，提出了以下三个目标：（1）我建议使用荧光报告基因进行整体成像，以生成形态发生过程的 4 维细胞描述。通过图像分析，我将全面重建参与耳囊泡形成的基板细胞的位置、轨迹和细胞分裂，并了解耳基板细胞如何从外胚层组织分层并重新排列成径向极化的空心组织。 (2) 我将使用有限元表示将耳囊建模为“加压壳”，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)我将模型与patterning的功能结合起来 跨上皮转运和细胞间粘附的基因。通过使用生物物理模型，我 将通过改变细胞间粘附蛋白表达和内淋巴压力来测试药理学和遗传扰动（突变体+变形体）的模型结果。这些扰动将证明细胞水平的模式电路如何控制形态发生过程的特定方面。这些目标将共同阐明模式输入如何通过控制内淋巴分泌和时空粘附蛋白表达来协调耳朵形态发生的细胞力学。