Shiqiu Li Dr. Scott Stewart Stankunas Lab UO SPUR 2012.

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Presentation transcript:

Shiqiu Li Dr. Scott Stewart Stankunas Lab UO SPUR 2012

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

Regeneration? Many mammals, like humans, cannot regenerate lost limbs Zebrafish can

What is the mechanism?

What initiates osteoblast differentiation and dedifferentiation?

WNT and BMP!

A Model ? What pathway is involved here? Transforming growth factor beta/Activin?

Activin or TGF-β II I Smad2 PP TGF-β and activin are ligands for pathway Both are part of TGF-β superfamily Both have a Type II and a Type I receptor TGF-β Type I receptor: Alk5 Activin Type I receptor: Alk4

Activin or TGF-β II I Smad4 Smad2 PP

Activin or TGF-β II I Smad4 Smad2 PP Transcription Regulation

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

Question What role do TGF-β/activin play in zebrafish fin regeneration? Hypothesis TGF-β/Activin signaling pathway is involved in cell proliferation in zebrafish fin regeneration

Early differentiation? Proliferation?

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

Methods Fixate and Dehydrate Xylene and paraffin Section Mount fin(s) Fluorescent microscopy Antibody staining Collect fish Amputate fish Wait Alk Inhibitor X Amputate fish Harvest fin(s)Short wait

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

Alk4 inhibitors cause reduced proliferation in osteoblasts Sp7 PCNA Sp7 PCNA Alk4iDMSOSp7 PCNA

Alk4 inhibitors cause reduced proliferation in osteoblasts P value =

So does TGF-β/activin induce cell proliferation of all cells in the blastema?

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

TGF-β/activin do not induce cell proliferation in the blastema of regenerating fin

Alk4 inhibitors cause reduced proliferation in osteoblasts P value =

Outline 1. Background 2. Question/Hypothesis 3. Methods 4. Earlier Data 5. Results 6. Conclusion/Finale

Conclusion The proliferation of cells in the blastema are not dependent on TGF-β/activin signaling. Osteoblast proliferation specifically may or may not be dependent on TGF-β/activin signaling due to differences of Alk4/5 inhibitor doses.

Future Directions Co-stain control and Alk4/5 inhibited fins with osteoblast markers such as Sp7 and Runx2 to fully determine TGF-β/activin’s role in osteoblast proliferation and early redifferentiation. Inhibit zebrafish with fibroblast growth factor receptor inhibitor and observe what happens using immunofuoresence.

Acknowledgements Peter O’Day Adam Unger UO SPUR NIH Stankunas Lab Dr. Scott Stewart Dr. Kryn Stankunas

Thank you for listening!!!