Effects of Mechanically-Induced Stress on the Proteome and Development

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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.