Role of Dopamine D2 Receptor in Wnt5a-mediated Dopaminergic Neuronal Development
- 주제(키워드) Wnt5a , dopamine D2 receptor , ERK
- 발행기관 고려대학교 대학원
- 지도교수 백자현
- 발행년도 2009
- 제출일 2009-01-03
- 학위수여년월 2009. 2
- 학위명 석사
- 학과 일반대학원 생명공학과
- 세부전공 의약생명공학 전공
- 원문페이지 45 p
- 실제URI http://www.dcollection.net/handler/korea/000000007218
- 본문언어 영어
- 제출원본 000045535436
초록/요약
Dopamine D2 receptor (D2R) is one of important receptors which are able to modulate neurophysiological functions in the CNS. Deficiency of D2R induces the lack of voluntary movement and akinesia. The well-known factor of Wnt5a as promoting dopaminergic neuronal development did not affect in D2 receptor knock-out mice. Co-treatment with both D2R agonist and Wnt5a increases the number of dopaminergic neurons more than in the case of only agonist or Wnt5a stimulation in WT mice. However the synergic effect of D2R agonist and Wnt5a in dopaminergic neuronal development was completely blocked by PD98059 (MAPK inhibitor) and haloperidol (D2R antagonist). Indeed, the result of immunoprecipitation of D2R and Wnt5a was shown that Wnt5a was associated with D2R. Dishevelled 3 (Dvl3) which is well-known as a signaling molecule of both canonical and non-canonical pathway of Wnt was phosphorylated by D2R agonist and Wnt5a in WT but not in D2R^(-/-) and this activation was also blocked by haloperidol. Nevertheless, phosphorylation of ERK was not affected by treatment of Wnt5a after knockdown of Dvl3. However, ERK activation induced by Wnt5a was blocked by PTX, LY294002 (PI3K inhibitor), and haloperidol. These findings suggest that the interaction between D2R and Wnt5a is a crucial role in dopaminergic neuronal development via PI3K and ERK pathway.
more초록/요약
도파민 제2형 수용체인 D2R은 중추신경계에서 도파민성 신경전달에 있어 신경 생리학적 기능을 조절하는 매우 중요한 역할을 담당한다. 도파민 제2형 수용체가 없는 쥐는 파킨슨씨 병과 유사하게 운동성이 느리거나 줄어있는 양상을 보이며, 도파민성 신경세포의 수가 줄어있는 경향성을 보인다. 도파민성 신경세포발달에 중요한 인자로 알려져 있는 Wnt5a는 D2R이 없는 쥐에서는 그 반응성이 나타나지 않았다. 또한 이러한 효과는 WT에서 D2R에 길항제인 haloperidol이나 MEK 억제제인 PD98059를 처리하였을 때 억제되었다. 실제적으로 Wnt5a는 ligand로서 D2R에 결합하고 있음을 확인하였고 이러한 결합으로 D2R을 통해 ERK가 인산화됨을 확인하였다. 이러한 관찰 결과들은 Wnt5a가 D2R을 통해 ERK를 활성화시켜 도파민성 신경세포의 발달을 유도하고 있음을 제시한다.
more목차
INTRODUCTION = 1
MATERIAL & METHODS = 4
1.Animal preparation and mesencephalic neuronal cell culture = 4
2.Immunocytochemistry = 5
3.Preparation of Wnt5a conditioned media = 5
4.Immunoprecipitation and Western blotting = 6
5.Ligand binding assay = 7
6.In situ hybridization = 8
7.Oligomeric siRNA transfection and drug treatments = 9
8.Western blot analysis of p-ERK = 10
9.Statistical analysis = 10
RESULTS = 11
1.The synergic effect on dopaminergic neuronal development through the D2R agonist and Wnt5a = 11
2.Interaction between Wnt5a and dopamine D2 receptor = 15
3.Effect of D2R agonist and Wnt5a on dishevelled3 phos-phorylation via dopamine D2 receptor = 18
4.Effect of Dvl3 expression knockdown in D2R-mediated ERK signaling by Wnt5a in dopaminergic mesencephalic neuronal culture = 20
5. PI3K-mediated ERK activation induced by Wnt5a = 25
DISCUSSION = 27
REFERENCES = 29
Abstract in Korean = 34
ACKNOWLEGEMENT = 35
