신경돌기발생에 있어서 Reelin-Dab1의 역할
Role of Reelin-Dab1 Signaling in the Neuritogenesis
- 주제(키워드) Reelin , Dab1 , Gαo , neuritogenesis , VLDLR , ApoER2
- 발행기관 아주대학교
- 지도교수 이영돈
- 발행년도 2016
- 학위수여년월 2016. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000021634
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Reelin is a secreted extracellular glycoprotein in the Cajal-Retzius cell that binds to the very low density lipoprotein receptor (VLDLR) and apolipoprotein E receptor 2 (ApoER2), inducing tyrosine phosphorylation of Dab1, a cytoplasmic adaptor protein of receptors. The Reelin-Dab1 signaling is regulatory key to migration and positioning of neuronal cell during brain development. It also reported that Reelin-deficient mice (reeler) showed a significant decrease in neurite formation and this depression was rescued by Reelin treatment. The neuritogenesis has been known to be promoted by the activation of Gαi/o signaling. Activated Gαi and Gαo were obviously localized in terminal portions of growing neuritis. Here, I study that a crosstalk between Reelin and Gαi/o signaling is involved in neurite formation. Reelin-induced neuritogenesis was suppressed by treatment of PTX and siRNA of Gαi and Gαο. Moreover, Reelin treatment induced activation of MAPK and JNK involved in the Gαi/o pathway. Also, in the precedent mechanism, the binding activity of Src to Dab1 and Gαo was prominently increased by Reelin. This result represents that neuritogenesis by Reelin facilitated through the activation of Gαi/o signaling. Also, expression of Reelin receptor, VLDLR is increased in the reeler mice than wild type mice at postnatal 3-14 days during the cerebellar development. But, ApoER2 is not changed. In addition, VLDL receptor in the cultured cerebellar cell is disappeared by Reelin treatment. This result represent clue about compensatory effect that is receptor gene expression through Reelin signal pathway
more목차
Part I. Role of Reelin-Dab1 signaling in the neuritogenesis
1. INTRODUCTION 2
2. MATERIALS AND METHODS 5
2.1 Animals 5
2.2 Cell Culture 5
2.3 Production of recombinant Reelin 6
2.4 Immunocytochemistry 7
2.5 Immunoblotting analysis 7
2.6 Immunoprecipitation 8
2.7 Measurement of neurite outgrowth and Statistical analysis 9
3. RESULTS 10
3.1 Depression of neurite outgrowth in Reelin- and Dab1-deficient mice 10
3.2 Decreased neurite formation in Reelin-deficient mice is recovered by the Reelin treatment 10
3.3 Suppressions of Reelin-induced neurite outgrowth by anti-Reelin antibody 13
3.4 Suppressions of Reelin-induced neurite outgrowth by Gαo/i inhibitor and Gαo siRNA. 13
3.5 Active Gαo in the neurite terminals is increased by Reelin 17
3.6 Reelin accelerates neuritogenesis by AKT-GSK3β and Gαo/i-MAPK-JNK pathway. 17
3.7 Dab1 and Gαo interact through Src 18
3.8 C3G and Rap1 is related crosstalk between Dab1 and Gαi/o 18
4. DISCUSSION 25
5. CONCLUSION 28
REFERENCES 29
PART Ⅱ. Compensatory effect of VLDLR expression on cerebellar development of Reelin-deficient mouse
1. INTRODUCTION 37
2. MATERIALS AND METHODS 39
2.1 Animals and genotyping 39
2.2 Immunocytochemistry 39
2.3 Reverse transcriptase polymerase chain reaction (RT-PCR) 40
2.4 Production of recombinant Reelin 40
2.5 Cerebellum neuron culture 41
3. RESULTS 42
3.1 VLDLR expression is predominant during development of P7 mouse cerebellar cortex 42
3.2 Expression of VLDLR and ApoER2 are co-localized in Purkinje cell body 42
3.3 Colocalization of Reelin and VLDLR or ApoER2 during cerebellar development 46
3.4 The level of VLDLR expression is higher in reeler than wild-type mice, but not ApoER2 46
3.5 VLDLR and ApoER2 levels during cerebellar development in yotari mice 46
3.6 VLDLR and ApoER2 expression in primary cultured neuron are induced by treatment of recombinant Reelin 47
4. DISCUSSION 53
5. CONCLUSION 56
REFERENCES 57
국문요약 62
