MRI: Acquisition of a Seahorse XFe 96 oxygen flux analyzer for integrative organismal and cellular biology

Colgate University, Hamilton NY

Investigators

Abstract

An award is made to Colgate University to undertake multi-disciplinary research at the cell and organelle level through the acquisition of a Seahorse XFe96 oxygen flux analyzer. This instrument, which facilitates detailed analysis of cellular metabolic rates, will support research in animal physiology, virology, genetics, and neurotransmitter biology. A diverse group of researchers across two primarily undergraduate institutions (PUI) are eager to incorporate the Seahorse XF analyzer within their classes and research programs. The requested instruments will play a central role in collaborative faculty-student research in biology and neuroscience at Colgate University and Hamilton College, and will facilitate current and future excellence in undergraduate research training in the sciences. In addition, the requested Seahorse instruments will have a significant impact on instructional laboratories across the curriculum in areas that include neuroscience, animal physiology, and microbiology. Between undergraduate research training and utilization in courses, this system will impact approximately 150 undergraduates a year. The projects will facilitate research with potential societal benefits in the fields of aging, viral infection, and cancer, and will provide outreach opportunities for high school students interested in STEM fields. The Seahorse XFe96 oxygen flux analyzer is a multimodal system capable measuring cellular metabolic parameters including glycolysis rate and mitochondrial stress. In addition to its use in classroom laboratory settings, this unique technology will be deployed in student-focused research laboratories to investigate the following research questions: (1) How do cellular metabolic rates change with body size and age of dogs and how are they linked to whole-animal metabolism? Differences in longevity demonstrate that small dogs have significantly longer lifespans that larger breed dogs. Thus, dogs constitute an informative group to study connections between life-history and physiology. We can only use Seahorse technology to untangle this basic biological rule due to the nature of working with primary fibroblast cells from young and old dogs. (2) How do reoviruses alter cellular metabolic pathways when used as oncolytic agents? Reoviruses are non-pathogenic mammalian viruses. However, very little is known about how reovirus replication alters a host cell's metabolic state, another basic biological principle that can only be elucidated with the use of a Seahorse XFe96 because metabolic alterations due to viral infections can only be measured using intact cells. (3) Does the transcription factor Krüppel-like factor 4 (KLF4) protect against cellular damage due to malfunctioning mitochondria, and KLF4 is a mediator of cell cycle regulation and development? However, the role of Klf4 in regulating cellular metabolic pathways remains to be elucidated. Seahorse XF technology is the only method to measure mitochondrial metabolism and glycolysis (as an alternative pathway). (4) How does the whole-animal cellular phenotype of intact C. elegans change when exposed to the estrogen-like molecule DES, which decreases fertility in animals? The Seahorse XF technology is perfect to determine the effects of DES on cellular metabolism over multiple generations. (5) What effect does ascorbic acid (AA) have on the growth and development of larvae of the tobacco hornworm, Manduca sexta? Larvae reared on AA-deficient diets grow less and develop more slowly relative to larvae fed a normal AA-containing diet. The mechanisms of AA-dependent growth in insects remain unclear and the seahorse technology will allow us to directly examine the metabolic underpinnings of this AA-dependent process. Together, these projects will significantly enhance student-faculty research at Colgate University and Hamilton College.