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