臺灣博碩士論文加值系統

直至今日為止，在醫學及臨床科學中，如何成功地幫助中樞神經系統的修復仍是一大挑戰。另外一方面，週邊神經系統之修復則提供了更多的可能性。倘若能對週邊神經系統修復的機制有更多地了解，也許對於日後中樞神經系統的修復能有更多助益。在這篇論文當中，我證實了橋接蛋白 SH2B1beta 可以成功地促進分化為類交感神經的大鼠腎上腺髓質嗜鉻細胞瘤細胞株 (簡稱 PC12 cells) 損傷後之修復。在修復受損的過程當中， SH2B1beta?n可以增強?nPKC 所調控之細胞遷移，以及 ERK1/2 和 AKT 所調控之神經軸再生進而促進 PC12 cells 的神經修復。除此之外，我也更進一步探討 SH2B1beta?n對海馬迴神經細胞修復的影響，目前的結果顯示， SH2B1beta 對海馬迴神經細胞的修復可能扮演著不同的角色。

Abstract...I
摘要...II
誌謝...III
Index...V
Abbreviations...VIII
Introduction...1
Materials and Methods...10
Reagents...10
Cell line and hippocampal neurons culture...11
Experimental injury models...12
Analysis of neurite re-growth...13
Analysis of cell migration...13
Immunoblotting and immunoprecipitation...14
Immunofluorescence staining...15
Statistical analysis...15
Results...16
Overexpression of SH2B1beta promotes wound healing of differentiated PC12 cells...16
Overexpression of SH2B1beta promotes neurite re-growth of differentiated PC12 cells...16
Phosphorylations of ERK1/2 and AKT are required for the neurite re-growth during wound healing...17
Overexpression of SH2B1beta enhances cell migration during wound healing...18
SH2B1beta enhances cell migration via PLCgamma1-PKC pathway...19
PKC activation promotes SH2B1beta-enhanced cell migration...20
Src may participate in SH2B1beta-elicited cell migration...21
Fine balance between cell migration and neurite outgrowth warrants better wound healing...22
SH2B1beta does not promote regeneration of hippocampal neurons...23
Discussion...25
Reference...28
Figures...38
Figure 1. Overexpression of SH2B1beta enhances wound healing of differentiated PC12 cells...38
Figure 2. Overexpression of SH2B1beta promotes neurite re-growth of differentiated PC12 cells after mechanical injury...40
Figure 3. Neurite re-growth during wound healing is ERK1/2-dependent...43
Figure 4. AKT-dependent neurite re-growth during wound healing of PC12 cells...44
Figure 5. Overexpression of SH2B1beta?nincreases cell migration during wound healing...46
Figure 6. Overexpression of SH2B1beta enhances phosphorylation of PLCgamma1 and PKC in PC12 cells...48
Figure 7. Inhibiting PKC activity decreases cell migration in PC12-SH2B1beta?ncells...50
Figure 8. PMA increases SH2B1beta-enhanced cell migration during wound healing...51
Figure 9. Overexpression of SH2B1beta enhances PMA-induced PKC phosphorylation and synthesis...52
Figure 10. Src may participate in PMA-mediated cell migration in PC12 cells...54
Figure 11. PMA treatment reduces wound closure and pERK1/2...55
Figure 12. Motility of cells overexpressing SH2B1beta is higher during healing days 0-4 and reduced during healing days 5-8...57
Figure 13. Overexpression of SH2B1beta does not promote neurite re-growth of hippocampal neurons...58
Figure 14. Overexpression of SH2B1beta does not promote cell migration of hippocampal neuron...60
Figure 15. Working model...62
Appendix...63
Figure A1. Dose-dependent inhibition of cell migration by PKC inhibition...63
Figure A2. Inhibition of PKC enhances pERK1/2 and wound closure...64
Figure A3. PKCdelta degradation is enhanced by PMA treatment during wound healing in PC12 cells...65
Figure A4. Src phosphorylation is up-regulated before wounding and down-regulated on healing day 7 in PC12-SH2B1beta cells...66

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