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목차
Abstract 12
제1장 서론 14
제1절 오피오이드 수용체의 생리적 기전 14
제2절 삼환계 항우울제의 기전과 부작용 22
제3절 비 선택적 양이온 통로 TRPC4 24
제2장 연구 재료 및 방법 28
제1절 세포 배양 및 발현 28
제2절 실험 용액 및 시약 29
제3절 단일세포 막전압 고정법 32
제4절 통계 및 유의성 평가 33
제3장 결과 34
제1절 오피오이드 수용체에 의한 TRPC4 이온통로의 활성 34
제2절 오피오이드 수용체에 의존적으로 TRPC4 이온통로의 활성을 유발하는 삼환계 항우울제 39
1. 삼환계 항우울제에 의한 TRPC4 이온통로의 억제 39
2. μOR에 의존적인 아미트립틸린에 의한 TRPC4 이온통로의 활성화 44
3. δOR와 κOR에 의존적인 삼환계 항우울제에 의한 TRPC4 이온통로의 활성화 48
4. 농도에 따라 TRPC 활성에 이상(biphasic) 효과를 보이는 삼환계 항우울제 55
5. Gi-pathway의 선택적 억제에 따른 삼환계 항우울제의 TRPC4 활성 효과 상실[이미지참조] 61
제3절 오피오이드 수용체의 삼환계 항우울제 결합 부위 69
1. 오피오이드 수용체의 리간드 결합 부위와 편향된 작용성 69
2. Asp 잔기의 돌연변이에 따른 아미트립틸린에 의한 TRPC4 활성 억제 71
제4장 고찰 85
참고문헌 90
Table 1. Substance of solution for patch clamping 30
Table 2. Chemical information 31
Figure 1. Distribution of opioid peptides and their receptors in the CNS 16
Figure 2. Types of opioid receptors 17
Figure 3. Types of G-protein signaling pathways 19
Figure 4. Biased agonism and functional selectivity of opioid agonists 20
Figure 5. Molecular structure of TCAs 23
Figure 6. Schematic representation of the TRP superfamily of ion channels 25
Figure 7. TRPC4β current is increased by DAMGO treatment depending on μOR co-expression. 36
Figure 8. TRPC4β current is increased by DPDPE treatment depending on δOR co-expression. 37
Figure 9. TRPC4β current is increased by U50488 treatment depending on κOR co-expression. 38
Figure 10. GTPγS-induced TRPC4β current is inhibited by AMI treatment. 41
Figure 11. GTPγS-induced TRPC4β current is inhibited by DES treatment. 42
Figure 12. GTPγS-induced TRPC4β current is inhibited by IMI treatment. 43
Figure 13. TRPC4β current is increased by AMI treatment depending on μOR co-expression. 45
Figure 14. TRPC4β current is increased by DES treatment depending on μOR co-expression. 46
Figure 15. TRPC4β current is increased by IMI treatment depending on μOR co-expression. 47
Figure 16. TRPC4β current is increased by AMI treatment depending on δOR co-expression. 49
Figure 17. TRPC4β current is increased by DES treatment depending on δOR co-expression. 50
Figure 18. TRPC4β current is increased by IMI treatment depending on δOR co-expression. 51
Figure 19. TRPC4β current is increased by AMI treatment depending on κOR co-expression. 52
Figure 20. TRPC4β current is increased by DES treatment depending on κOR co-expression. 53
Figure 21. TRPC4β current is increased by IMI treatment depending on κOR co-expression. 54
Figure 22. TRPC4β current is not affected by 0.1 μM AMI treatment, regardless of μOR co-expression. 56
Figure 23. TRPC4β current is increased by 1 μM AMI treatment depending on μOR co-expression. 57
Figure 24. TRPC4β current is increased by 10 μM AMI treatment depending on μOR co-expression 58
Figure 25. TRPC4β current is not affected by 50 μM AMI treatment, regardless of μOR co-expression 59
Figure 26. The increase in TRPC current due to MOR modulation exhibits a biphasic effect, dependent on the concentration of AMI. 60
Figure 27. PTX pre-treatment blocks AMI-induced activation of TRPC4, which was modulated by μOR modulation. 63
Figure 28. PTX pre-treatment blocks AMI-induced activation of TRPC4, which was modulated by δOR modulation. 64
Figure 29. PTX pre-treatment blocks AMI-induced activation of TRPC4, which was modulated by κOR modulation. 65
Figure 30. Dominant negative mutant of Gαi₂ inhibits AMI-induced activation of TRPC4, which was modulated by μOR modulation.[이미지참조] 66
Figure 31. Dominant negative mutant of Gαi₂ inhibits AMI-induced activation of TRPC4, which was modulated by δOR modulation.[이미지참조] 67
Figure 32. Dominant negative mutant of Gαi₂ inhibits AMI-induced activation of TRPC4, which was modulated by κOR modulation.[이미지참조] 68
Figure 33. The sequence alignment of OR subtypes 70
Figure 34. The μOR(D147R) mutant did not alter DAMGO-induced TRPC4 activation. 73
Figure 35. The δOR(D128R) mutant did not alter DPDPE-induced TRPC4 activation. 74
Figure 36. The κOR(D138R) mutant did not alter U50488-induced TRPC4 activation. 75
Figure 37. The increase in TRPC4 current triggered by AMI did not occur in μOR(D147R). 76
Figure 38. The increase in TRPC4 current triggered by AMI did not occur in δOR(D128R). 77
Figure 39. The increase in TRPC4 current triggered by AMI did not occur in κOR(D138R). 78
Figure 40. The μOR(D147R) mutation induces diminished activation of TRPC4 by AMI. 79
Figure 41. The δOR(D128R) mutation induces diminished activation of TRPC4 by AMI. 80
Figure 42. The κOR(D138R) mutation induces diminished activation of TRPC4 by AMI. 81
Figure 43. The μOR(D147R) mutation induces diminished activation of TRPC4 by DAMGO. 82
Figure 44. The δOR(D128R) mutation induces diminished activation of TRPC4 by DPDPE. 83
Figure 45. The κOR(D138R) mutation induces diminished activation of TRPC4 by U50488. 84