Immunophilin ligands and GDNF enhance neurite branching or elongation from developing dopamine neurons in culture
Immunophilin ligands and GDNF enhance neurite branching or elongation from developing dopamine neurons in culture
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DOI：
10.1006/exnr.2000.7417
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发表时间：
2000-07-01
影响因子：
5.3
通讯作者：
Isacson, O
中科院分区：
文献类型：
--
作者：
Costantini, LC;Isacson, O
免疫亲和素配体的神经营养作用已经在外周和中枢神经系统损伤的动物模型中显示出来。为了研究这些化合物的特异性促生长作用，我们检测了不同的免疫亲和素配体对原代培养的多巴胺(DA)神经元的影响，并与众所周知的DA营养因子-胶质细胞系源性神经营养因子(GDNF)进行了比较。在胚胎第14天的中脑腹侧神经元培养中，免疫亲和素配体促进了DA突起的延长，与载体处理组相比，这抑制了钙调神经磷酸酶(FK506和环孢素A)的活性。GDNF也观察到了这种延长，已知通过磷酸化依赖的途径发挥其营养作用。相反，不抑制钙调神经磷酸酶的免疫亲和素配体(雷帕霉素和V-10,367)增加了DA神经突起的分支，这表明延长依赖于维持的磷酸化，而分支不依赖。此外，V-10,367和雷帕霉素都拮抗FK506的延长效应和诱导的分支。伸长的拮抗作用(和分支的重现)说明了发育中的DA神经元伸长或分支的内在能力，但不是两者兼而有之。我们证明了免疫亲和素FKBP12(12-kDa FK506结合蛋白)在腹侧中脑神经元培养中表达并与DA神经元共定位，本工作阐明了GDNF和免疫亲和素配体通过与细胞内靶点的相互作用来调节DA神经元发育生长过程的特定促生长作用，(C)2000学术出版社。
Neurotrophic effects of immunophilin ligands have been shown in animal models of peripheral and central nervous system insult. To investigate the specific growth-promoting effects of these compounds, we examined the effects of various immunophilin ligands on primary dopamine (DA) neurons in culture and compared these with a well-known DA trophic factor, glial cell line-derived neurotrophic factor (GDNF). In neuronal cultures from Embryonic Day 14 ventral mesencephalon, enhanced elongation of DA neurites was observed with immunophilin ligands, which inhibited the phosphatase activity of calcineurin (FK506 and cyclosporin A) when compared to vehicle-treated cultures. This elongation was also observed with GDNF, known to exert its trophic effects through phosphorylation-dependent pathways. In contrast, immunophilin ligands that do not inhibit calcineurin (rapamycin and V-10,367) increased branching of DA neurites, suggesting that elongation is dependent upon maintained phosphorylation while branching is not, In addition, both V-10,367 and rapamycin antagonized the elongation effects of FK506 and induced branching. The antagonism of elongation (and reappearance of branching) illustrates the intrinsic abilities of developing DA neurons to either elongate or branch, but not both, We show that the immunophilin FKBP12 (12-kDa FK506-binding protein) is expressed in ventral mesencephalic neuronal cultures and colocalizes with DA neurons, This work elucidates the specific growth-promoting effects by which GDNF and immunophilin ligands modify developmental growth processes of DA neurons, via their interactions with intracellular targets, (C) 2000 Academic Press.