2026-09-22T18:58:15Zhttps://repository.ajou.ac.kr/oai/request

oai:repository.ajou.ac.kr:201003/130442024-10-11T02:21:26Zcom_201003_14372com_201003_14368col_201003_14374

Regulation of Astrocyte Functions by a Parkinson's Disease Gene, DJ-1 파킨슨병 유전자 DJ-1에 의한 성상세포 기능 조절 연구 최, 동주 조, 은혜 대학원 의생명과학과 110942 최, 동주 Parkinsons's disease DJ-1 Astrocyte Astrogliosis Anti-inflammatory function INTRODUCTION . 1 A. Parkinson's disease (PD) 1 B. Genes associated with PD . 2 1. DJ-1 (PARK7) 2 2. Other genes . 3 C. Function of astrocytes in the normal brain . 6 D. Function of astrocytes in the injured brain . 7 1. Reactive astrocytes : intermediate filaments (Ifs) and morphological features . 7 2. Reactive astrocytes : function of astrogliosis in neural protection . 8 3. Reactive astrocytes : function of astrogliosis for regeneration and repair . 9 4. Reactive astorycytes : functin of astrogliosis for regulation of inflammation . 11 E. Astrocyte dysfunction in neurodegenerative disease 12 F. Aim of this study 13 II. Materials and methods 15 1. Ethics statement . 15 2. DJ-1 deficient mice . 15 3. Animal MRI 15 4. Stereotaxic surgery and drug injection . 16 5. Tissue preparation . 16 6. Immunohistochemistry 17 7. Histological quantification 18 8. Organotypic cortical slice cultures 18 9. Cell culture 19 10. Western blot analysis . 19 11. Immunoprecipitation 21 12. Quantitative real-time PCR. 21 13. Reverse transcriptase PCR 22 14. DNA constructs . 23 15. Transfection 23 16. Proximity ligation assay (PLA) 23 17. Small interfering RNAs (siRNA) 24 18. Prostaglandin D2 (PGD2) ELISA . 25 19. Inhibition of protein synthesis by cycloheximide 25 20. Inhibition of Ubiquitination by MG132 25 21. Measurement of nitric oxide 25 22. Statistical analysis . 26 III. RESULTS 27 Part A. DJ-1, (PARK7), plays critical roles in astrogliosis by regulating of the pSTAT3 signaling pathway for tissue repair after brain injury 27 1. Repair of brain injury was attenuated in DJ-1 KO mice 27 2. DJ-1 deficiency causes a defect of astrogliosis . 30 3. Reduction of astrocyte derived GDNF expression and attenuated restoration of dopaminergic processes in the DJ-1 KO brain 41 4. DJ-1 rescued insufficient astrogliosis in DJ-1 KO brain slice cultures. 47 5. STAT3 activation (tyrosine phosphorylation of STAT3, pSTAT3) was attenuated in DJ- 1 KO slices 50 Part B. DJ-1 exerts the anti-inflammatory effects of astrocytes through stabilization of Sox9, a transcription factor that regulates lipocalin-type prostaglandin D2 synthase (L-PGDS) expression. 52 1. DJ-1 deficiency caused a defect of anti-inflammatory function of astrocytes. 52 2. DJ-1 deficiency caused a defect of PGD2 expression by attenuation of L-PGDS in astrocytes. 57 3. DJ-1 deficiency caused a defect of Sox9 protein expression by regulation of ubiquitination in astrocytes. 65 IV. DISCUSSION 73 V. SUMMARY AND CONCLUSION. 85 REFERENCES. 86 국문요약 . 109 Doctor 2016-11-29T06:08:37Z 2016-11-29T06:08:37Z 2016 2016 Thesis Theses http://repository.ajou.ac.kr/handle/201003/13044 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000022796 000000022796 en application/pdf