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目的:探讨转录因子垂体同源盒家族因子3(Pitx3)和孤儿核受体相关因子1(Nurr1)在多巴胺(DA)能神经元终末分化中的表达特点。方法:采用免疫荧光方法检测体外培养的中脑源性神经干细胞(mNSCs)诱导分化后和大鼠胚胎发育至出生腹侧中脑Pitx3、Nurr1和酪氨酸羟化酶(TH)的表达。结果:(1)体外培养的mNSCs诱导分化48 h的Map-2阳性细胞不表达Pitx3、Nurr1和TH;分化7 d的TH阳性细胞均表达转录因子Pitx3和Nurr1;(2)在大鼠腹侧中脑黑质(SN)、腹侧被盖区(VTA)和中缝背核(DRN)可见大量TH与Pitx3(或Nurr1)共表达的神经元;(3)Pitx3和Nurr1阳性细胞主要分布于E16.5、P0大鼠SN、VTA和DRN区,其中Pitx3阳性细胞还少量分布于腹侧中脑非DA能神经元区域,Nurr1阳性细胞在腹侧中脑分布范围较Pitx3更广泛。结论:转录因子Pitx3、Nurr1在中脑DA能神经元的终末分化和生存维持中起重要作用。
Objective: To investigate the expression of Pitx3 and ornuclear receptor 1 (Nurr1) in the terminal differentiation of dopaminergic neurons. Methods: The expression of Pitx3, Nurr1 and Tyrosine hydroxylase (TH) in cultured ventral midbrain after induced by mMNCs and cultured in vitro were detected by immunofluorescence. Results: (1) Pitx3, Nurr1 and TH were not expressed in Map-2 positive cells induced by mNSCs cultured for 48 h in vitro. The transcription factors Pitx3 and Nurr1 were expressed in TH positive cells on the 7th day after differentiation. (2) A large number of neurons co-expressed with TH and Pitx3 (or Nurr1) were found in SN, VTA and DRN. (3) Pitx3 and Nurr1 positive cells mainly distributed in E16 .5, P0 rats SN, VTA and DRN area, Pitx3 positive cells were also distributed in the ventral mesencephalic non-DA neurons, Nurr1 positive cells in the ventral mesencephalon distribution range broader than Pitx3. Conclusion: The transcription factors Pitx3 and Nurr1 play an important role in the terminal differentiation and survival of DA neurons in midbrain.