GDNF signaling in embryonic midbrain neurons in vitro
GDNF signaling in embryonic midbrain neurons in vitro
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DOI：
10.1016/j.brainres.2007.04.071
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发表时间：
2007-07-23
期刊：
影响因子：
2.9
通讯作者：
di Porzio, Umberto
中科院分区：
文献类型：
--
作者：
Consales, Claudia;Volpicelli, Floriana;di Porzio, Umberto
胶质细胞源性神经营养因子（GDNF）对许多细胞类型，包括中脑多巴胺能（mDA）神经元发挥营养作用。使用大鼠中脑原代培养丰富的mDA神经元，我们表明，延长GDNF刺激增加他们的生存和轴突生长。它调节DA功能（酪氨酸羟化酶，TH和多巴胺转运蛋白，dat）和DA高亲和力摄取所必需的基因的表达。为了鉴定参与GDNF信号通路的基因，我们使用了GDNF刺激3小时的mDA培养物的DNA微阵列。在这里，我们表明，GDNF信号顺序激活基因编码的转录因子EGR 1和TIEG。此外，它增加了cav 1的表达，cav 1编码小窝的主要成分。GDNF还调节编码钙调磷酸酶亚基ppp 3R 1和ppp 3CB的基因的表达，并抑制钙-钙调蛋白依赖性蛋白激酶II β亚型（CaMK II β）基因表达。这些蛋白质参与神经元分化和突触可塑性。此外，GDNF刺激下调了参与神经元凋亡的糖原合成酶激酶3 β（gsk 3 β）基因的表达。使用特定的细胞内信号转导通路的抑制剂，我们发现egr 1，tieg，cav 1，CaMk II β和gsk 3 β基因表达的变化是细胞外信号调节激酶1/2（ERK）依赖的，而cAMP依赖的蛋白激酶（PKA）途径影响ppp 3R 1和ppp 3CB基因表达的上调。这些结果表明，GDNF刺激导致参与神经元可塑性和存活以及mDA功能的基因的转录调节，部分由ERK和PKA信号转导介导。(C)2007 Elsevier B. V.保留所有权利。
The glial cell line-derived neurotrophic factor (GDNF) exerts trophic actions on a number of cell types, including mesencephalic dopaminergic (mDA) neurons. Using rat mesencephalic primary cultures enriched in mDA neurons, we show that protracted GDNF stimulation increases their survival and neurite outgrowth. It modulates the expression of genes essential for DA function (tyrosine hydroxylase, TH and dopamine transporter, dat) and of DA high affinity uptake. To identify genes involved in GDNF signaling pathways, we have used DNA microarray on mDA cultures stimulated with GDNF for 3 h. Here we show that GDNF signaling sequentially activates the genes encoding for the transcription factors EGR1 and TIEG. In addition, it increases the expression of cav1, which encodes for the major component of caveolae. GDNF also modulates the expression of the genes encoding for the Calcineurin subunits ppp3R1 and ppp3CB, and inhibits calcium-calmodulin-dependent protein kinase II beta isoform (CaMKII beta) gene expression. These proteins are involved in neuronal differentiation and synaptic plasticity. Moreover, GDNF stimulation down regulates the expression of the glycogen synthase kinase 3 beta (gsk3 beta) gene, involved in neuronal apoptosis. Using inhibitors of specific intracellular signal transduction pathways we show that changes of egr1, tieg, cav1, CaMkII beta and gsk3 beta genes expression are extracellular-signal regulated kinases 1/2 (ERK)-dependent, while the cAMP-dependent protein kinase (PKA) pathway influences the up-regulation of ppp3R1 and ppp3CB gene expression. These results demonstrate that GDNF stimulation results in the transcriptional modulation of genes involved in neuronal plasticity and survival and in mDA function, mediated in part by ERK and PKA signaling. (C) 2007 Elsevier B.V. All rights reserved.