In vivo analysis of mechanotransduction

力转导的体内分析

• 批准号: 10456813
• 负责人: Erin Jean Cram
• 金额: $ 33.46万
• 依托单位: NORTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10456813
• 关键词: 4D Imaging; Address; Asthma; Biochemical; Biological; Biophysics; Biosensor; Blood Vessels; Caenorhabditis elegans; Calcium; Cell Shape; Cells; Cellular biology; Communication; Computer Models; Contractile System; Contracts; Cyclic AMP-Dependent Protein Kinases; Embryo; Engineering; GTP-Binding Protein alpha Subunits; GTP-Binding Protein alpha Subunits, Gs; Gap Junctions; Genetic Models; Heart Diseases; Heterotrimeric GTP-Binding Proteins; Hypertension; Image; Individual; Knowledge; Life; Link; Lung; Mammary gland; Measures; Mechanics; Modeling; Molecular Genetics; Nematoda; Oocytes; Phospholipids; Physiologic pulse; Process; Proteomics; Regulation; Reproducibility; Reproductive system; Research; Role; Salivary Glands; Signal Transduction; Smooth Muscle; Stretching; Structure; System; Testing; Time; Tissues; Translating; Tube; Tubular formation; Urine; Uterus; Work; automated image analysis; cell type; computerized tools; filamin; gene network; genetic manipulation; in vivo; in vivo Model; insight; lymphatic vessel; mechanical signal; mechanotransduction; novel; pressure; reproductive tract; response; rho GTPase-activating protein; spatiotemporal

In vivo analysis of mechanotransduction Cells in biological tubes must integrate biochemical and mechanical cues in order to expand or contract in a coordinated manner. Inappropriate responses to changing states underlie conditions such as heart disease, hypertension and asthma. Despite insights from biophysics and from cell biology on engineered substrates, many important questions remain regarding how mechanical information is sensed by cells, translated into biochemical signals, and integrated to produce a coordinated tissue-level response. For example, how is multicellular contractility regulated in space and time? How are the induction and propagation of biochemical signals regulated by mechanical cues? How do different cell types within a tissue coordinate their actions? To address these questions, we have developed an in vivo model, the C. elegans spermatheca, which is a tubular tissue in the nematode reproductive system comprised of 24 smooth-muscle-like cells that connect to the uterus via a toroidal valve. The major advantages of this system are that the cells are naturally stretched and contract as oocytes enter, and are amenable to quantitative live imaging and targeted genetic manipulation, enabling observation and manipulation of individual cells in the context of an intact tissue. We have discovered that oocyte entry induces Ca2+ pulses that sweep across the tissue, culminating in a coordinated contraction that pushes the fertilized embryo into the uterus. Ca2+ release and contractility in the spermatheca and valve are coordinated such that while the spermathecal bag contracts, the valve dilates to allow exit of the fertilized embryo. Well-conserved gene networks regulate these processes, suggesting broad applicability of our findings to other contractile systems. Here, we propose a combination of 4D imaging of genetically-encoded biosensors, proteomics, molecular genetics, and modeling to elucidate the mechanisms which coordinate Ca2+ signaling in response to stretch. Specifically, we will 1) test the hypothesis that the heterotrimeric G protein, Gαs, signals through PKA to regulate spermathecal contractility; 2) model the mechanisms by which stretch triggers calcium release and signal propagation; and 3) determine how valve contractility is regulated, both autonomously and via communication from the spermathecal bag. This research will lead to important advances in our understanding of the fundamental mechanisms by which cells convert mechanical information into biochemical signals, and how this signaling is integrated to regulate tissue function.

机械转移的体内分析 生物管中的细胞必须整合生化和机械提示，以扩展或 以协调的方式收缩。对不断变化的国家的不适当回应是基础 心脏病，高血压和哮喘等疾病。尽管有生物物理学的见解 从工程基质的细胞生物学中，关于 细胞如何感知机械信息，转化为生化信号，并 集成以产生协调的组织级反应。例如，多细胞如何 收缩性在时间和时间上受到监管？生化的诱导和传播如何 机械提示调节的信号？组织坐标中的不同细胞类型如何 他们的行动？为了解决这些问题，我们已经开发了一种体内模型，即秀丽隐杆线虫 精子，它是线虫复制系统中的一个管状组织，已完成24 光滑肌肉样细胞通过环形阀连接到子宫。主要优势 该系统的是，当卵母细胞进入时，细胞是自然拉伸的，并且是收缩的，并且是 适合定量实时成像和靶向遗传操作，使观察能够观察 并在完整组织的背景下操纵单个细胞。我们发现 卵母细胞进入诱导Ca2+脉冲扫过组织，并在协调中 收缩将受精的胚胎推入子宫。 CA2+释放和收缩性 精子和瓣膜是协调的，使精子袋合同时， 瓣膜扩张以允许施肥的胚胎退出。保存良好的基因网络调节这些 过程，表明我们的发现对其他收缩系统的广泛适用性。在这里，我们 提案的遗传编码生物传感器，蛋白质组学，分子的4D成像组合 遗传学和建模以阐明协调Ca2+信号传导的机制 对拉伸的响应。具体而言，我们将1）检验以下假设：异三聚体G蛋白， GαS，通过PKA信号调节精子收缩力； 2）通过 拉伸会触发钙释放和信号传播； 3）确定阀门如何 自主和通过精子袋的通信进行了监管。 这项研究将导致我们对基本的理解的重要进展 细胞通过将机械信息转换为生化信号的机制，以及如何 该信号被整合以调节组织功能。