shiqiu li dr. scott stewart stankunas lab uo spur 2012
TRANSCRIPT
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Transforming Growth Factor-Beta/Activin and Zebrafish Fin Regeneration: Induction of
Cell Proliferation?Shiqiu Li
Dr. Scott StewartStankunas LabUO SPUR 2012
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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Regeneration?Many mammals, like humans, cannot
regenerate lost limbsZebrafish can
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What is the mechanism?
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What initiates osteoblast differentiation and dedifferentiation?
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WNT and BMP!
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A Model?
What pathway is involved here?
Transforming growth factor beta/Activin?
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ActivinorTGF-β
II ISmad
2P P
• 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
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ActivinorTGF-β
II I
Smad4
Smad2 P P
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ActivinorTGF-β
II I
Smad4
Smad2 P P
Transcription Regulation
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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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
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Early differentiation?
Proliferation?
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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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
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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Alk4 inhibitors cause reduced proliferation in osteoblasts
Sp7PCNA
Sp7PCNA
Alk4iDMSO Sp7PCNA
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Alk4 inhibitors cause reduced proliferation in osteoblasts
× 100
P value = 0.0049
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So does TGF-β/activin induce cell proliferation of all cells in the blastema?
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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TGF-β/activin do not induce cell proliferation in the blastema of regenerating fin
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TGF-β/activin do not induce cell proliferation in the blastema of regenerating fin
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Alk4 inhibitors cause reduced proliferation in osteoblasts × 100
P value = 0.9941
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Outline1. Background2. Question/Hypothesis3. Methods4. Earlier Data5. Results6. Conclusion/Finale
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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.
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Future DirectionsCo-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.
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AcknowledgementsPeter O’DayAdam UngerUO SPURNIHStankunas Lab
Dr. Scott StewartDr. Kryn Stankunas
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Thank you for listening!!!