Effects of Mechanically-Induced Stress on the Proteome and Development

这笔赠款将支持更好地了解物理环境如何影响生物学的工作。 具体来说，这笔赠款将支持更好地了解蛋白质如何对机械力作出反应的工作。 生物体不断地感知机械力并做出适当的反应。 例如，地球的引力不断向下拉，大气压力从各个方向压缩，但生物体可以保持直立。如果感知这些物理效应的能力受到损害，可能会导致发育障碍，肌肉和骨骼损失，神经异常，免疫和年龄相关问题。 即使在宇航员身上也观察到了这些情况，他们在远离地球的长时间内暴露在微重力下。 在这项资助下完成的工作使用了模拟不同物理环境的新型微技术系统。 这些微技术系统将用于研究物理环境中的机械变化如何差异地影响数千种蛋白质。 确定这些生物反应的特征将最终导致国民健康的改善。例如，这项工作的结果可能最终为未来的治疗策略或疾病预防技术提供信息。该项目还将为跨学科合作创建一个教育和培训计划。 工程师和生物科学家将共同努力，代表性不足的群体参与研究的范围将扩大。地球上不同类型的物理力量不断地影响着生物，这是它们不断适应外部刺激的一部分。虽然对特定感觉系统中的机械传导了解很多，但对整个生物体对周围物理环境的广义机械感觉知之甚少。这种知识缺乏的一个原因是有限的方法来机械刺激足够的整个生物体，使全面的蛋白质组学分析。为了研究广义机械感觉，数百个活的果蝇胚胎将暴露于多种模式，振幅和持续时间的各种机械刺激，如微重力，超重力和外部压缩，使用微制造的高通量设备，将开发。这项工作将利用2D-DIGE（2维差异凝胶电泳）的比较蛋白质组学方法，以确定响应于这些不同模式的机械刺激的蛋白质丰度或翻译后调节的变化。这种方法还将使局部/急性与慢性/普遍存在的机械刺激的蛋白质组学比较成为可能。通过使用比较蛋白质组学与发育表型的分析相结合，这些机械模式的专业化程度，或在整个生物体的背景下机械转导的通用途径重叠将被检查。该奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。

This grant will support work to better understand how the physical environment influences biology. Specifically, this grant will support work to better understand how proteins respond to mechanical forces. Organisms are continuously sensing mechanical forces and respond appropriately. For example, the Earth’s gravity is constantly pulling down and the atmospheric pressure which compresses from all directions, yet organisms can remain upright. If the ability to sense these physical effects is compromised, it can lead to developmental disorders, muscle and bone loss, neurological abnormalities, immunological, and age-related problems. These conditions have been observed even in astronauts, who are exposed to microgravity during prolonged times away from Earth. The work done under this grant use novel microtechnology systems that mimic different physical environments. These microtechnology systems will be used to investigate how mechanical changes in the physical environment differentially affect thousands of proteins. Characterizing these biological responses will ultimately lead to improvements in national health. For example, the results of this work may ultimately inform future therapeutic strategies or disease prevention techniques. This project will also create an educational and training program for interdisciplinary collaboration. Engineers and biological scientists will work together, and participation in research for underrepresented groups will be broadened. Different types of Earth’s physical forces continuously impact living beings as a part of their ceaseless adaptation to external stimuli. While much is known about mechanotransduction in specific sensory systems, little is known about generalized mechanosensation of the surrounding physical environment by whole organisms. One reason for this lack of knowledge is the limited methods to mechanically stimulate enough whole organisms to enable comprehensive proteomic analysis. To investigate generalized mechanosensation, hundreds of living Drosophila embryos will be exposed to multiple modes, amplitudes, and durations of various mechanical stimulation such as microgravity, hypergravity, and external compression using microfabricated high-throughput devices that will be developed. This work will utilize the comparative proteomics approach of 2D-DIGE (2 Dimensional Difference Gel Electrophoresis) to identify changes in protein abundance or post-translational regulation in response to these different modes of mechanical stimulation. This approach will also enable the proteomic comparison of local/acute to chronic/ubiquitous mechanical stimulation. By using comparative proteomics in conjunction with the analysis of developmental phenotypes, the extent to which these mechanical modes are specialized, or overlap in a universal pathway of mechanotransduction in the context of whole organisms will be examined.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.