1. Zebrafish Heart Regeneration Occurs by Cardiomyocyte Dedifferentiation and Proliferation Yajun Lin (Jocelyn) Circulation Research August 4, 2006; 99 (3)

2. Outline Heart Disease – heart failure Heart Regeneration Green Fluorescent Protein Mechanistic Insight of Zebrafish Heart Regeneration

3. Heart Disease CDC, National Vital Statistics System and the U.S. Census Bureau.

4. Heart Regeneration in Human Mercader et al.European Society of Cardiology (2013)

5. Zebrafish Heart Regeneration Auman et al. PLoS Biol.: 2007, 5(3);e53 PMID:17311471PMID:17311471

6. Mechanistic Insight of Zebrafish Heart Regeneration Research done by Jopling et al. Barcelona, Spain.

7. Preparation Transgenic zebrafish were generated in which 4-OHT- inducble Cre in under the control of the cardiomyocyte- specific cmlc2a promoter. The reporter construct used the same cmlc2a promoter followed by a floxed stop cassette and enhanced green fluorescent protein. Removal of the stop cassette by Cre allows the cmlc2a promoter within the transgene to drive the expression of GFP.

8. Green Fluorescent Protein

9. Regenerated cardiomyocytes are derived from differentiated cardiomyocytes. C Jopling et al. Nature 464, 606-609 (2010) doi:10.1038/nature08899

10. Differentiated cardiomyocytes re-enter the cell cycle. C Jopling et al. Nature 464, 606-609 (2010) doi:10.1038/nature08899

11. Cardiomyocytes dedifferentiate, resulting in the disassembly of sarcomeric structure and detachment. C Jopling et al. Nature 464, 606-609 (2010) doi:10.1038/nature08899

12. plk1 is necessary for cardiac regeneration. C Jopling et al. Nature 464, 606-609 (2010) doi:10.1038/nature08899

13. Conclusion Zebrafish heart regeneration is driven primary by pre- existing cardiomyocytes The vest majority of heart muscle cell arise frim pre-existing cardiomyocytes Pre-existing cardiomyocytes undergo limited dedifferentiation. Cardiomyocytes close to the wound start to disassemble their sarcomeric structure, re-entering the cell cycle. Many sarcomeric gene being downregulated after amputation.

14. References Heidi J Auman, Hope Coleman, Heather E Riley, Felix Olale, Huai-Jen Tsai, Deborah Yelon Functional modulation of cardiac form through regionally confined cell shape changes. PLoS Biol.: 2007, 5(3);e53 PMID:17311471 PMID:17311471 Laflamme, Michael A, and Charles E Murry. 2011. "Heart regeneration." Nature 473, no. 7347: 326-335. The 2006 Holiday Lectures Series "Potent Biology: Stem Cells, Cloning, and Regeneration.” Jopling, Chris, Eduard Sleep, Marina Raya, Mercè Martí, Angel Raya, and Juan Carlos Izpisúa Belmonte. 2010. "Zebrafish heart regeneration occurs by cardiomyocyte dedifferentiation and proliferation." Nature 464, no. 7288: 606-609. O. Shimomura, FH Johnson, and Y Saiga (1962). Extraction, Purification and Properties of Aequorin, a Bioluminescent Protein from the Luminous Hydromedusan, Aequorea. J. Cellular and Comparative Physiol. 59: 223-239 Roger Heim, Andrew B. Cubitt & Roger Y. Tsien (1995) Improved Green Fluorescence. Nature. 373: 663-664.

15. The End