Ca2+ oscillations induced by testosterone enhance neurite outgrowth
Ca2+ oscillations induced by testosterone enhance neurite outgrowth
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DOI：
10.1242/jcs.02775
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发表时间：
2006-02-15
影响因子：
4
通讯作者：
Ehrlich, BE
中科院分区：
文献类型：
--
作者：
Estrada, M;Uhlen, P;Ehrlich, BE
替吉奥在调节神经元功能方面具有短期和长期作用。在这里，我们展示了快速的细胞内雄激素受体的非依赖性作用，睾酮对细胞内钙在神经母细胞瘤细胞。我们确定了睾酮诱导的Ca 2+信号，主要开始于神经突尖端，随后向细胞核传播，然后重复以产生振荡模式。最初的瞬变依赖于生产肌醇1，4，5-三磷酸[Ins（1，4，5）P-3]，但随后的瞬变需要细胞外Ca 2+内流和Ca 2+从细胞内存储释放。抑制百日咳毒素敏感的G蛋白受体或使用针对Ins（1，4，5）P-3受体1型的siRNA阻断了Ca 2+反应，而抑制或敲低细胞内雄激素受体则没有效果。细胞溶质和细胞核Ca 2+用小清蛋白缓冲，所述小清蛋白被工程化以靶向细胞溶质或细胞核。细胞质小清蛋白阻断Ca 2+信号在两个隔室，核小清蛋白只阻断核信号。突变体小清蛋白的表达没有改变睾酮诱导的Ca 2+信号。神经母细胞瘤细胞中的神经突生长通过添加睾酮而增强。这种作用被抑制时，胞质Ca 2+缓冲和衰减时，小白蛋白靶向细胞核。我们的研究结果与睾酮的快速作用一致，涉及Ins（1，4，5）P-3介导的Ca 2+振荡，并支持以下观点：用于神经元细胞分化的途径中存在协同作用，涉及快速非基因组作用和雄激素的经典基因组作用。
Testosterone has short- and long-term roles in regulating neuronal function. Here, we show rapid intracellular androgen receptor-independent effects of testosterone on intracellular Ca2+ in neuroblastoma cells. We identified testosterone-induced Ca2+ signals that began primarily at the neurite tip, followed by propagation towards the nucleus, which was then repeated to create an oscillatory pattern. The initial transient depended upon production of inositol 1,4,5-trisphosphate [Ins(1,4,5)P-3], but subsequent transients required both extracellular Ca2+ influx and Ca2+ release from intracellular stores. Inhibition of pertussis toxin-sensitive G-protein receptors or the use of siRNA for the Ins(1,4,5)P-3 receptor type 1 blocked the Ca2+ response, whereas inhibition or knock-down of the intracellular androgen receptor was without effect. Cytosolic and nuclear Ca2+ were buffered with parvalbumin engineered to be targeted to the cytosol or nucleus. Cytoplasmic parvalbumin blocked Ca2+ signaling in both compartments; nuclear parvalbumin blocked only nuclear signals. Expression of a mutant parvalbumin did not modify the testosterone-induced Ca2+ signal. Neurite outgrowth in neuroblastoma cells was enhanced by the addition of testosterone. This effect was inhibited when cytosolic Ca2+ was buffered and was attenuated when parvalbumin was targeted to the nucleus. Our results are consistent with a fast effect of testosterone, involving Ins(1,4,5)P-3-mediated Ca2+ oscillations and support the notion that there is synergism in the pathways used for neuronal cell differentiation involving rapid non-genomic effects and the classical genomic actions of androgens.