2012 Annual Report
1a.Objectives (from AD-416):
The objective of this research area is to develop novel imaging technologies aimed at confronting critical issues facing production animal agriculture by monitoring, in real-time, cellular and molecular processing in the context of the living organism. Specific research projects will cover a broad range of research in the cellular and molecular biological sciences, disease-environment interactions, animal-plant interfaces, and growth and developmental physiology with applications aimed at understanding physiological mechanisms with a specific emphasis on enhancing production performance in livestock.
1b.Approach (from AD-416):
As part of this initiative, novel technologies which utilize the photon (light), thermal signatures (heat), spectroscopy and fluorescence will be adapted to cellular- and molecular-based strategies to permit physiological processes to be monitored in a dynamic fashion at the levels of single, living cells to entire organisms in vivo. These non-invasive technologies (e.g., biophotonics, using light as a quantitative indicator beacon ofmolecular events) will enable the expression of genes, the invasiveness of bacteria, the breakdown of plant or dietary components, or hormone-receptor interactions to be visualized in living systems both in the laboratory and field, and under traditional livestock production environments. Faculty with expertise in functional imaging will interface with collaborating scientists working in the animal, plant and veterinary sciences to develop these novel systems aimed at addressing specific hypothesis-driven and production-based questions. Results from this initiative will not only develop new models to advance scientific progress in reproductive biology, food safety, disease, plant-animal interactions, and environmental physiology, but will also develop technological advancements that will address experimentally critical questions which heretofore have not been addressable in living systems. Finally, we will expand the use of biophotonic-based technologies to address physiological questions in animals with potential application to field-based monitoring systems.
Specific accomplishments from 2011-2012 include advancing our biophotonic imaging capabilities. In addition, advancements in molecular approaches in bio-marker identification and continued development of gamete and embryo systems for developmental monitoring are all significant research directions achieved this reporting period.
New laboratory animal models have been continued to speed development of biophotonic paradigms for livestock-based applications. These have been accomplished using amphibian model paradigms which allow us to use larger egg and embryo systems to test feasibility of our developmental monitoring approaches.
Biomarkers are being identified through bioinformatic approaches for oocytes and embryos using proteomic approaches, protein-receptor studies and gene transcription-biophotonic paradigms.
Thermal imaging technologies remain a focus of our research, being applied to the monitoring of (1) reproductive function in livestock; (2) mammary physiology in dairy cattle; and (3) measures of thermal comfort (e.g., energetics) and/or stress-related responses among livestock and other species (e.g., avian) in the production or natural environment. This aspect of the program is being highlighted at the Smithsonian’s Folklife Festival from June 27 – July 8, 2012, with an exhibit on the mall in Washington DC.
In summary, accomplishments under the Biophotonics Initiative as applied to agricultural livestock, some peripheral applications for model development are yielding new research tools which have the potential to develop translatable technologies for an enhanced understanding of physiological processes to improve agricultural livestock production, health and/or overall profitability.
This report documents research as appropriated from the Senate Report of the Agriculture, Rural Development, Food and Drug Administration, and Related Agencies Appropriations Bill (7 USC Sec. 3101) for salaries and expenses of the Agricultural Research Service under the committee recommendation heading “Biotechnology Research to Improve Crops and Livestock” as a Specific Cooperative Agreement with the USDA-ARS Catfish Genetics Research Program at Stoneville, MS “Biophotonics - the Application of Novel Imaging Methodologies to Livestock Production Research. (Project Number: 6402-21310-001-01)”.
Elucidating spermatozoan markers that may influence developmental processes. Equine spermatozoa are highly sensitive to cryopreservation and not all stallion semen freezes alike. Unfortunately, there are no reliable markers to predict the freezability status of stallions, which would enhance the widespread application of cryopreservation in the equine industry. The present study aimed at discriminating potential “bad” and “good” freezer stallions in a herd.
Developing new biophotonic models. Quantitative Bioluminescence imaging of functional estrogen receptor activity within intact porcine ovarian follicles in vitro. Activated estrogen receptors (ER) in response to estrogen bind to specific sequence called estrogen response elements (ERE) and induce transcription. The objective of these investigations were to evaluate whether the estrogen induced ER binding activity in granulosa cells of antral ovarian follicles is determined by bioluminescence imaging system, and how estrogen concentrations in follicular fluid affect the ER binding activity. Through our research, scientist at Mississippi State University, have demonstrated the development of a new methodology for measuring functional and ligand activated estrogen receptors in intact porcine ovarian follicles in vitro using a bioluminescence molecular imaging system.
Developing new biophotonic models. Self-illuminating quantum dots are nanoparticles that are less than 100 nanometers in diameter. Their coating with the light-emitting protein Renilla luciferase forms complexes which have promising applications in in vivo imaging. These complexes can be further combined to specific tags such as antibodies or peptides for various in vitro studies. Especially in reproduction, these conjugates may contribute to a better comprehension of molecular events associated with fertilization and beyond. To this end, scientist at Mississippi State University, evaluated the ability of mammalian spermatozoa to harmlessly incorporate nanoparticles.
Elucidating oocyte and embryo markers. Mammalian oocytes progressively acquire their full developmental competency or quality within the growing follicle. It is well-known that poor quality oocytes are generally derived from small compared to large follicles. Additionally, oocytes matured in the presence of follicular fluid of different follicle sizes display distinct developmental competency. Follicular fluid (FF) contains various molecules whose composition is influenced by external factors having beneficial or harmful effects on oocyte quality. Full characterization and evaluation of FF predisposition to such factors would make FF optimal sources for detecting suitable predictors of oocyte quality. In our studies, scientist at Mississippi State Univeristy, compared the proteome profiles of FF at different follicular developmental stages.
Sykes, D.J., Couvillion, J.S., Cromiak, A., Bowers, S., Schenck, E., Crenshaw, M., Ryan, P. 2012. The use of digital infrared thermal imaging to detect estrus in gilts. Theriogenology. 78:147-152.
Greene, J.M., Dunaway, C.M., Bowers, S.D., Rude, B.J., Feugang, J.M., Ryan, P. 2012. Dietary L-arginine supplementation during mouse gestation enhances reproductive performance and Vegfr2 transcription activity in the fetoplacental unit. Journal of Nutrition. 142(3):456-460.
Scott, V.L., Shack, L.A., Eells, J.B., Ryan, P.L., Donaldson, J.R., Coats, K.S. 2012. Immunomodulator expression in trophoblasts from the feline immunodeficiency virus (FIV)- infected cat. Journal of Virology. 8:336.
Feugang, J.M., Willard, S.T., Ryan, P.L. 2012. In vitro manipulation of mammalian gametes and embryos: What are we learning from animal settings? Journal of Fertilization:In Vitro. 2:114.