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Role of Sonic Hedgehog in Development of the Central Nervous System

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dc.contributor.author박, 래희-
dc.date.accessioned2011-11-07T04:43:40Z-
dc.date.available2011-11-07T04:43:40Z-
dc.date.issued2011-
dc.identifier.urihttp://repository.ajou.ac.kr/handle/201003/4369-
dc.description.abstractSonic hedgehog (Shh) functions as a morphogen to induce early dorso-ventral patterning of the central nervous system (CNS) and specifies cell fates of spinal motor neurons. Shh has also been known to promote cell cycle progression of embryonic stem cells as well as adult neural stem/progenitor cells. However, it has not been elucidated how Shh induces neural induction during early development of nervous system.

To elucidate the role of Shh in neural induction, we used a pluripotent embryonal carcinoma cell line, P19 and investigated the effects of Shh on inducing the expression of nestin, the marker for neural stem/progenitor cells. In early stages, we found that Shh promoted the CNS-specific enhancer of the nestin gene through regulation of E-box on nestin 2nd intron 5’-region by Nmyc that is a target gene of Shh signaling. Shh also increased population and proliferation of neural stem cells in vitro or in vivo. In late stages, Shh induced specification of motor neurons as well as dopaminergic neurons. Simultaneously, Shh inhibited the expression of meso-endoderm lineage markers, Gsc, Sox17, and GATA4. The results suggest that Shh promotes neurogenesis in multiple ways: by inducing the neuroectodermal cells, by repressing non-neural cell fates, and by promoting differentiation of neurally committed progenitor cells into motor and dopaminergic neurons.
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dc.description.tableofcontentsI. 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
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dc.language.isoen-
dc.titleRole of Sonic Hedgehog in Development of the Central Nervous System-
dc.title.alternative신경발생과정에서 Shh의 역활-
dc.typeThesis-
dc.identifier.urlhttp://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000011545-
dc.subject.keywordSonic hedgehog-
dc.subject.keywordNeural induction-
dc.subject.keywordDifferentiation-
dc.subject.keywordNestin-
dc.subject.keywordN-myc-
dc.subject.keywordP19 cells-
dc.subject.keywordPro-neural gene-
dc.subject.keywordNeurogenin1-
dc.subject.keywordMash1-
dc.description.degreeDoctor-
dc.contributor.department대학원 의학과-
dc.contributor.affiliatedAuthor박, 래희-
dc.date.awarded2011-
dc.type.localTheses-
dc.citation.date2011-
dc.embargo.liftdate9999-12-31-
dc.embargo.terms9999-12-31-
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