|PROPOSAL NUMBER:||03- II B1.03-9171|
|SUBTOPIC TITLE:||Bioscience and Engineering|
|PROPOSAL TITLE:||Low-power, Confocal Imaging of Protein Localization in Living Cells (7215-010)|
PRINCIPAL INVESTIGATOR/PROJECT MANAGER
(Name, E-mail, Mail Address, City/State/Zip, Phone)
Anthony A Ferrante
20 New England Business Center
Andover, MA 01810-1077
U.S. Citizen or Legal Resident: Yes
TECHNICAL ABSTRACT (LIMIT 200 WORDS)
The proposed innovative technology addresses the need to understand and develop countermeasures for the skeletal and cardiovascular changes to astronauts during prolonged exposure to microgravity environments. During the proposed program we will apply our innovative functional imagery approach, developed in Phase I research, to the problem of microgravity-induced cellular changes in osteoblasts, cells that are responsible for bone growth and repair. As part of the program we will develop a 593-nm diode-pumped, solid-state (DPSS) laser that will be combined with a commercially available 473 nm DPSS laser to enable simultaneous visualization of three cellular proteins that are genetically fused to fluorescent reef coral proteins (RCFPs). The genetic constructs we will generate, integrin aV-ZsGreen, p130CAS-DsRed2 and HcRed-a-actin, will enable space biologists to monitor changes in cytoskeletal structure as well as changes at the focal adhesions in vivo and in real time with no manipulation or reagent addition. Those same genetic constructs could be expressed in different cell lines to examine the effects of microgravity environments on cardiovascular tissue. We expect that the improved understanding of cellular changes that our innovative model system will deliver will enable rapid development of countermeasures to microgravity-induced changes in humans.
POTENTIAL NASA COMMERCIAL APPLICATIONS (LIMIT 100 WORDS)
We anticipate that the successful completion of the Phase II program will have several non-NASA commercial implications. First, we expect that this work will increase the adoption of the RCFPs by terrestrial researchers in many fields. The RCFPs will provide enhanced ability for simultaneously monitoring expression of multiple genes. We also anticipate that the 593-nm laser that will be developed during the program will be adopted by manufacturers and sellers of confocal imaging systems. There is currently no commercially available, solid-state laser with an emission wavelength between 532-nm and 635-nm.
POTENTIAL NON-NASA COMMERCIAL APPLICATIONS (LIMIT 100 WORDS)
We anticipate that space biology researchers at NASA will use the osteoblast cell culture model and associated confocal imaging system that will be developed during our Phase II program for flight experiments aimed at understanding changes at the cellular level that lead to the skeletal changes found in astronauts during long-term space travel. We further anticipate that the knowledge gained will enable rapid development of effective countermeasures for microgravity-induced bone loss. In addition, the genetic constructs and the confocal imaging system may also provide enhanced capabilities for understanding the cellular mechanisms of cardiovascular changes found in astronauts.