Michael Gutkin (Biology)
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CELLULAR EVENTS IN ZEBRAFISH OPTIC TECTAL BRAIN EXPLANTS: A MODEL TO ANALYZE NEUROTRAUMA AND
NEUROREGENERATION
by
Michael C. Gutkin
Reflective Tutorial in Biology(Senior Learning Community)
Experiential Component: ResearchDepartment of Biological Sciences
Wagner College
Spring, 2010
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RESEARCH FOCUSES IN THE LABORATORY
• To study cellular events taking place in nervous tissue in cases of Traumatic Brain Injury (TBI).
• To use a model animal whose brain has known regenerative capacities.
• To use a model animal which inexpensive and easy to handle.
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ZEBRAFISH (DANIO RERIO)
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ZEBRAFISH BRAIN
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CULTURE CONDITIONS
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• Light microscopic analysis showed formation of embryoid structures as early as 48 hours in culture which further differentiated with the extended time in culture. This picture is from a sample cultivated for 96 hours.
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MY PERSONAL RESEARCH GOALS
• Scanning electron microscopy
• Confocal laser scanning microscopy
• BrdU Cell Proliferation• Immunohistochemistry
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SCANNING ELECTRON MICROSCOPYTOP ROW – 12 HOURS OF CULTIVATION
BOTTOM ROW – 24 HOURS OF CULTIVATION
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SCANNING ELECTRON MICROSCOPY48 HOURS OF CULTIVATION
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SCANNING ELECTRON MICROSCOPY96 HOURS OF CULTIVATION
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ZEBRAFISH BRAIN CYTOARCHITECTURE SEEN WITH IMMUNOHISTOCHEMISTRY AND CONFOCAL
MICROSCOPY
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BRDU ASSAY FOR CELL PROLIFERATION6 & 48 HOURS OF CULTIVATION
6 H
48 H
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BRDU ASSAY FOR CELL PROLIFERATION96 HOURS OF CULTIVATION
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BRDU ASSAY FOR CELL PROLIFERATIONPOSITIVE CONTROL: CHO CELLS
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CONCLUSION
• Adult zebrafish brain demonstrates regenerative capacities in surviving organotypic culture.
• This regeneration is characterized in SEM by relocation of “stem-like cells” and the formation of embryoid bodies accompanied by neovascularization within spongiform degenerative regions.
• Vital cells can be detected by IHC as well as cell proliferation, but dividing cells are seen less than expected.
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ACKNOWLEDGEMENTS
• Professor Christopher Corbo • Dr. Alejandra del C. Alonso • Dr. William L’Amoreaux• Dr. Zoltan Fulop• Professor Linda Raths