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신경발생과정에서 Shh의 역활
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 서해영 | - |
| dc.contributor.author | Park, Rae Hee | - |
| dc.date.accessioned | 2019-10-21T07:17:29Z | - |
| dc.date.available | 2019-10-21T07:17:29Z | - |
| dc.date.issued | 2011-02 | - |
| dc.identifier.other | 11545 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/17849 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의학과,2011. 2 | - |
| dc.description.tableofcontents | I. INTRODUCTION 1 PART I 1 A. Hh(hedgehog) 1 B. The role of Hh signaling in neural 2 C. Dorso-ventral patterning 4 D. Cell type specification 5 E. Cell proliferation in tumors 6 F. P19 embryonic carcinoma cells (EC) 7 G. Purpose 8 PART II 9 A. Neural stem cells 9 B. Nestin 9 C. basic helix-loop-helix (bHLH) transcription factor 10 II. MATERIALS AND METHODS 12 A. Materials 12 B. Mathods 13 1. Plasmids 13 2. P19 cell culture 13 3. Differentiation of P19 cells 14 3.1 Conventional differentiation 14 3.2 Current differentiation 14 4. Reverse Transcription Polymerase Chain Reaction (RT-PCR) 15 5. Western blotting 16 6. Luciferase assay 17 7. RNA interference 17 8. Immunocytochemistry 18 9. In utero injection and electroporation 18 10. Immunohistochemistry 19 11. Chromatin Immunoprecipitation (ChIP) assay 20 III. RESULTS 22 PART I 22 1. Shh induces neural lineage and blocks meso-endodermal line in P19 cells 22 2. Shh regulates CNS specific Nestin expression through Nestin 2nd intron in P19 cells 24 3. Shh directly promtes CNS specific Nestin expression in P19 cells 27 4. Shh promotes CNS specific Nestin expression through N-myc 29 5. Shh induces population of neural stem cells in vivo 34 6. Shh directly induces neural differentiation of P19 cells 38 7. Shh promotes differentiation of P19 cells into motor and dopaminergic neurons 40 8. Shh regulates the expression of Id and Hes1 during neural differentiation of P19 cells 42 9. Id expression correlates with stemness of NSCs 44 10. Id proteins increase the proliferating neural precursors in the developing brain 46 PART. II 51 11. Ngn1 regulates CNS specific nestin expression though the NE2 of Nestin 2nd intron3-region 51 12. Ngn1 promotes neural progenitor cells into glutamatergic neuron in vivo 57 IV. DISCUSSION and CONCLUSIONS 66 REFERENCES 72 국문요약 83 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 신경발생과정에서 Shh의 역활 | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.department | 일반대학원 의학과 | - |
| dc.date.awarded | 2011. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 569276 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011545 | - |
| dc.subject.keyword | Sonic hedgehog | - |
| dc.subject.keyword | Neural induction | - |
| dc.subject.keyword | Differentiation | - |
| dc.subject.keyword | Nestin | - |
| dc.subject.keyword | N-myc | - |
| dc.subject.keyword | P19 cells | - |
| dc.subject.keyword | Pro-neural gene | - |
| dc.subject.keyword | Neurogenin1 | - |
| dc.subject.keyword | Mash1 | - |
| dc.description.alternativeAbstract | Nestin is known as a specific marker for neural stem cells and regulated by the enhancer located on the nestin 2nd intron of nestin gene. Nestin 2nd intron is highly conserved among species and crucial for nestin expression in central nervous system (CNS) progenitor cells. The transgenic mice that constructed to express green fluorescence protein (GFP) with nestin enhancer have been developed and have great advantage to represent live neural stem cells during research in vivo and vitro. In this study, we characterized GFP expressing the neural stem cells or progenitors in vitro culture. We also investigated that Ngn1, which is a basic helix-loop-helix (bHLH) transcription factor that is expressed in neuronal precursors during development of the nervous system, can promote nestin expression by nestin 2nd intron enhancer. E-box 2 in 637bp of nestin 2nd intron were thought to be strong candidates for bHLH factors during the early neurogenesis. Ngn1 increased the E-box 2 mediated reporter gene activity. To evaluate the functional mechanism of E-box mediated nestin gene expression in vivo, we generated transgenic (Tg) mice with the 637bp fragment of nestin 2nd intron containing the wild or mutated E-box2. Wild type tg mice successfully exhibited CNS-specific LacZ expression. We also found that LacZ positive cells are proliferating neural progenitors or intermediate progenitors, which is expressed with Ki67 and Tbr2, and can differentiate into vGLUT2 positive glutamatergic neurons in developing cortex. These results indicate that pro-neural Ngn1 promotes nestin expression through nesitn 2nd intron E-box 2 and Ngns mediated LacZ positive cells can differentiate into glutamatergic neurons. | - |
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