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
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.
ZEBRAFISH (DANIO RERIO)
ZEBRAFISH BRAIN
CULTURE CONDITIONS
• 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.
MY PERSONAL RESEARCH GOALS
• Scanning electron microscopy
• Confocal laser scanning microscopy
• BrdU Cell Proliferation• Immunohistochemistry
SCANNING ELECTRON MICROSCOPYTOP ROW – 12 HOURS OF CULTIVATION
BOTTOM ROW – 24 HOURS OF CULTIVATION
SCANNING ELECTRON MICROSCOPY48 HOURS OF CULTIVATION
SCANNING ELECTRON MICROSCOPY96 HOURS OF CULTIVATION
ZEBRAFISH BRAIN CYTOARCHITECTURE SEEN WITH IMMUNOHISTOCHEMISTRY AND CONFOCAL
MICROSCOPY
BRDU ASSAY FOR CELL PROLIFERATION6 & 48 HOURS OF CULTIVATION
6 H
48 H
BRDU ASSAY FOR CELL PROLIFERATION96 HOURS OF CULTIVATION
BRDU ASSAY FOR CELL PROLIFERATIONPOSITIVE CONTROL: CHO CELLS
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.
ACKNOWLEDGEMENTS
• Professor Christopher Corbo • Dr. Alejandra del C. Alonso • Dr. William L’Amoreaux• Dr. Zoltan Fulop• Professor Linda Raths
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