Activity-dependent release of transforming growth factor-beta in a neuronal network in vitro
Activity-dependent release of transforming growth factor-beta in a neuronal network in vitro
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DOI：
10.1016/j.neuroscience.2007.09.046
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发表时间：
2007-12-12
期刊：
影响因子：
3.3
通讯作者：
Krieglstein, K.
中科院分区：
文献类型：
--
作者：
Lacmann, A.;Hess, D.;Krieglstein, K.
对于神经营养素以及转化生长因子β（TGF-β）家族的成员，已经提出了合成和释放的活性依赖性调节。结合观察到神经递质的分泌是由神经营养因子启动的，可以合理地假设它们可能充当逆行调节剂，增强突触的功效和稳定性。在本研究中，我们测试了这一假设，并以胚胎E16.5的小鼠原代海马神经元为模型，研究了体外TGF-β的释放和调节。我们表明，神经元活性在体外调节TGF-β的释放和TGF-β的表达。根据 MLEC/纤溶酶原激活剂抑制剂 (PAI) 荧光素酶测定的评估，用 KCI、3-藜芦碱（藜芦定）、谷氨酸或氯化氨甲酰胆碱（卡巴胆碱）处理培养物会增加分泌的 TGF-β 水平，而在河豚毒素或河豚毒素存在的情况下，由 KCI 或藜芦定刺激的 TGF-β 释放会减少。 1,2-双(2-氨基苯氧基)乙烷-N,N,N',N'-四乙酸(BAPTA)。此外，谷氨酸盐的应用显着上调了培养物中TGF-β2和TGF-β3的表达。值得注意的是，KCI 刺激导致 Smad（SMA（线虫）和 MAD=mothers against dpp（果蝇）的复合术语）易位到细胞核中，并上调 TGF-β 诱导早期基因 (Tieg1) 的表达，表明活性依赖性释放的 TGF-β 可能发挥自分泌作用，从而激活 TGF-β 依赖性信号通路。总之，这些结果表明体外小鼠海马神经元中 TGF-β 的活性依赖性释放和基因转录，以及海马网络内释放的 TGF-β 的后续自分泌功能。 (c) 2007 年国际广播组织。由爱思唯尔有限公司出版。保留所有权利。
For neurotrophins and also for members of the transforming growth factor beta (TGF-beta) family an activity-dependent regulation of synthesis and release has been proposed. Together with the observation that the secretion of neurotransmitters is initiated by neurotrophic factors, it is reasonable to assume that they might act as retrograde modulators enhancing the efficacy and stabilization of synapses. In the present study, we have tested this hypothesis and studied the release and regulation of TGF-beta in vitro using mouse primary hippocampal neurons at embryonic day E16.5 as model.We show that neuronal activity regulates TGF-beta release and TGF-beta expression in vitro. Treatment of the cultures with KCI, 3-veratroylveracevine (veratridine), glutamate or carbamylcholine chloride (carbachol) increased the levels of secreted TGF-beta, as assessed by the MLEC/plasminogen activator inhibitor (PAI)luciferase-assay, whereas TGF-beta release stimulated by KCI or veratridine was reduced in the presence of tetrodotoxin or 1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid (BAPTA). In addition, application of glutamate significantly upregulated expression of TGF-beta 2 and TGF-beta 3 in the culture. Notably, KCI stimulation caused Smad (composite term from SMA (C. elegans) and MAD=mothers against dpp (Drosophila)) translocation into the nucleus and upregulated TGF beta inducible early gene (Tieg1) expression, demonstrating that activity-dependent released TGF-beta may exert autocrine actions and thereby activate the TGF-beta-dependent signaling pathway. Together, these results suggest an activity-dependent release and gene transcription of TGF-beta from mouse hippocampal neurons in vitro as well as subsequent autocrine functions of the released TGF-beta within the hippocampal network. (c) 2007 IBRO. Published by Elsevier Ltd. All rights reserved.