MRI: Acquisition of a Fluorescence Stereomicroscope to Enhance Undergraduate Research at Colorado College

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This Major Research Instrumentation grant supports the acquisition of a fluorescence stereomicroscopeand an imaging system to enhance the research programs of Dr. Darrell J. Killian and Dr. Nancy N.Huang in the Biology Department at Colorado College. The real-time visualization of fluorescentmolecules in living model organisms has become an indispensable technique because it allows the directobservation and documentation of changes in gene activity, the localization of proteins within a cell, andthe tracking of cells and tissues during development. An important question in biology is how changes ingene activity determine sex-specific development. The Killian Lab is interested in the identification andcharacterization of genes that affect sex-specific development in the popular model organismCaenorhabditis elegans (C. elegans), a soil-dwelling roundworm. C. elegans populations contain malesand hermaphrodites, which have some obvious morphological differences. However, sex-specific celltype differences in the nervous system are not easy to see. Fluorescent markers for sex-specific celltypes have emerged as powerful and sensitive tools for the identification of defects in sex-specificdevelopment. Mutants have been isolated that are morphologically normal males or hermaphrodites, butwith neurons that are normally only found in the opposite sex. The identification of the genes affected bythese mutations and their subsequent characterization will provide new insights into the genetic basis ofsex-specific development. Fluorescence stereomicroscopy and imaging are essential for the observationand documentation of sexual defects that are detected in vivo using fluorescent markers for sex-specificcell types. The Huang Lab is interested in how cellular identity, or cell fate specification, is established.MEX-3 is a protein that plays an important role in cell fate specification during C. elegans embryonicdevelopment. The MEX-3 protein is initially present in all cells of the embryo but becomes depleted fromthe posterior cells by the four-cell stage. This change in MEX-3 distribution is required for theestablishment of the anterior/posterior axis of the embryo, thus it is important to understand how MEX-3 islocalized in the embryo. Preliminary evidence suggests that MEX-3 is actively degraded in a manner thatis dependent upon two different types of protein modifications termed phosphorylation and ubiquitination.The identification of the specific sites of these modifications within the MEX-3 protein and theidentification of genes that regulate these modifications are of great interest. Fluorescence microscopywill enable an in vivo study of the stability of fluorescently tagged MEX-3 when specific sites in MEX-3 aremutated or when other genes in the C. elegans genome are disrupted.Colorado College is the only nationally ranked liberal arts college in the Rocky Mountain region of theUnited States and enrolls approximately 2000 undergraduates from all over the United States andinternationally. The Biology Department has a strong commitment to conducting original research withundergraduate collaboration and the fluorescence microscope and imaging system will provide importantresearch and training opportunities for approximately 6-10 undergraduates working in the Killian andHuang labs each year. In addition, many courses in the Biology curriculum feature inquiry-basedresearch lab experiences. Students with such experiences are best equipped to handle any postgraduatechallenges that they might face. The equipment will augment and improve laboratory activities,demonstrations, and experiments associated with several courses and will impact approximately 200undergraduates per year. The imaging system will be a particularly valuable teaching tool because it willenable the instructor and a group of students to simultaneously observe the same sample on a computermonitor. The Biology Department also plays a strong service role in the college by offering non-majorscourses that foster a strong appreciation for basic biology research and empower students to makeinformed decisions and opinions on many popular topics in biology such as stem cell research, genetherapy, genetically modified organisms, and environmental issues. The fluorescence stereomicroscopewill provide non-majors with unique and exciting opportunities to observe cutting-edge science first-hand.Finally, Colorado College actively seeks to enhance diversity on campus and the availability of state-ofthe-art instrumentation, such as a fluorescence stereomicroscope and imaging system, will help attractunderrepresented minorities and women to participate in biology research.