Brain-derived neurotrophic factor signalling in adult pig retinal ganglion cell neurite regeneration in vitro
Brain-derived neurotrophic factor signalling in adult pig retinal ganglion cell neurite regeneration in vitro
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DOI：
10.1016/j.brainres.2004.02.023
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发表时间：
2004-05-08
期刊：
影响因子：
2.9
通讯作者：
Hicks, D
中科院分区：
文献类型：
--
作者：
Bonnet, D;Garcia, M;Hicks, D
脑源性神经营养因子（BDNF）在啮齿类动物模型的体内和体外刺激视网膜神经节细胞（RGC）存活和轴突再生中有牵连，但关于神经营养因子在高等哺乳动物中的作用的数据很少。因此，我们分析了成年猪RGC的原代培养物中的BDNF信号传导。通过免疫组化、HPLC分析和RT-PCR检测，BDNF蛋白和mRNA在体内和体外的猪视网膜中均有表达，可能参与了RGC神经突的发生。最初的剂量反应研究确定了最佳效果由20 ng/ml BDNF诱导，导致神经突长度增加约3倍。我们分析了磷脂酰肌醇3激酶（PI 3 K）和丝裂原活化蛋白激酶（MAPK）级联对BDNF诱导的神经突起再生的各自贡献。添加PI 3 K抑制剂渥曼青霉素或MAPK抑制剂U 0126阻断了50-100%的BDNF诱导的神经突伸长; U 0126也显著降低了神经突再生，低于未处理的对照水平。trk受体抑制剂K252 a对神经突再生或形态学没有可观察到的影响。因此，这些数据表明，BDNF是一个强大的刺激剂的神经突起生长的RGC制备从成年大型哺乳动物视网膜，至少有两个信号通路的因果关系。基于BDNF的治疗可能在治疗人类RGC变性中具有潜在的用途。(C)2004 Elsevier B. V.保留所有权利。
Brain-derived neurotrophic factor (BDNF) has been implicated in stimulating retinal ganglion cell (RGC) survival and axonal regeneration in rodent animal models in vivo and in vitro, but very little data are available on neurotrophin effects in higher mammals. We hence analysed BDNF signalling in primary cultures of adult pig RGC. As detected by immunohistochemistry, HPLC analysis and RT-PCR, BDNF protein and mRNA were present within pig retina in vivo and in vitro, where it may be involved in baseline RGC neuritogenesis. Initial dose-response studies established optimal effects were induced by 20 ng/ml BDNF, leading to an approximately threefold increase in neurite length. We analysed the respective contributions of phosphatidyl inositol 3 kinase (PI3K) and mitogen activated protein kinase (MAPK) cascades to BDNF-induced neurite regeneration. Addition of either the PI3K inhibitor wortmannin or the MAPK inhibitor U0126 blocked 50-100% BDNF-induced neurite elongation; U0126 also significantly reduced neurite regeneration below untreated control levels. The trk receptor inhibitor K252a had no observable effect on neurite regeneration or morphology. These data hence demonstrate that BDNF is a potent stimulator of neurite growth in RGC prepared from an adult large mammal retina, and that at least two signalling pathways are causally involved. BDNF-based therapy may be of potential use in treating RGC degeneration in hunians. (C) 2004 Elsevier B.V. All rights reserved.