This document details research on cellular events in zebrafish optic tectal brain explants as a model for studying traumatic brain injury and neuroregeneration. The study finds that adult zebrafish brain exhibits regenerative characteristics, including the relocation of 'stem-like cells' and formation of embryoid bodies over time in culture. The research employs various microscopy techniques to analyze cell proliferation and regeneration in this model organism.

1. CELLULAR EVENTS IN ZEBRAFISH OPTIC TECTAL BRAIN EXPLANTS: A MODEL TO ANALYZE NEUROTRAUMA AND NEUROREGENERATION by Michael C. GutkinReflective Tutorial in Biology(Senior Learning Community) Experiential Component: ResearchDepartment of Biological SciencesWagner College Spring, 2010

2. Research Focuses in the LaboratoryTo 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.

3. Zebrafish (Daniorerio)

4. Zebrafish Brain

5. Culture Conditions

6. 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.

7. My Personal Research GoalsScanning electron microscopy Confocal laser scanning microscopy BrdU Cell ProliferationImmunohistochemistry

8. Scanning Electron MicroscopyTop Row – 12 Hours of cultivationBottom Row – 24 Hours of Cultivation

9. Scanning Electron Microscopy48 Hours of Cultivation

10. Scanning Electron Microscopy96 Hours of Cultivation

11. Zebrafish Brain Cytoarchitecture seen with Immunohistochemistry and Confocal Microscopy

12. BrdU Assay for Cell Proliferation6 & 48 Hours of Cultivation6 h48 h

13. BrdU Assay for Cell Proliferation96 Hours of Cultivation

14. BrdU Assay for Cell ProliferationPositive Control: CHO Cells

15. ConclusionAdult 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.

16. Acknowledgements Professor Christopher CorboDr. Alejandra del C. Alonso Dr. William L’AmoreauxDr. ZoltanFulopProfessor Linda Raths