Effect of FGF-9 on cultured neurons : novel approach for the treatment of neurodegenerative disorders

FGF-9 对培养神经元的影响：治疗神经退行性疾病的新方法

• 批准号: 10557060
• 负责人: KANDA Takashi
• 金额: $ 5.82万
• 依托单位: Tokyo Medical and Dental University Graduate School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557060/
• 关键词: FGF-9; Alzheimer disease; basal forebrain cholinergic neuron; autocrine; paracrine; amyotrophic lateral sclerosis; blood-brain barrier; neurotrophic factor; NGF; ALS

We examined the effect of fibroblast growth factor (FGF)-9 on primary cultures of rat basal forebrain cholinergic neurons (BFCN) obtained at embryonic day 17 and anterior horn cholinergic neurons (AHCN) obtained embryonic day 14. FGF-9 enhanced survival of AChE-positive neurons, increased their mean soma size, and upregulated choline their acetyltransferase (ChAT) activity in BFCN and AHCN.For BFCN, The ChAT-promoting effect of FGF-9 was approximately as potent as that of nerve growth factor (NGF) and was greater than those of basic fibroblast growth factor (bFGF), ciliary neurotrophic factor (CNTF), or glia-derived neurotrophic factor (GDNF). Simultaneous addition of FGF-9 and NGF induced extremely high ChAT levels, suggesting that FGF-9 and NGF may enhance cholinergic properties in BFCN via different pathways that can act synergistically. In immunocytochemical and in situ hybridization studies in cultured cells and also in sections of adult rat brain, BFCN showed cytoplasmic immunostaining for FGF-9 and expressed FGF-9 messenger RNA ; thus, we concluded that FGF-9 acts on BFCN in an autocrine and/or paracrine manner. Thus, we conclude that FGF-9 may be a promising candidate for therapeutic trials in Alzheimer disease and amyotrophic lateral sclerosis.The next step is to develop effective methods to deliver exogenous FGF-9 into the central nervous system. We coupled FGF-9 with some polyamines and this FGF-9-polyamine complex can pass through in vitro BBB system ; however, the neurotrophic effect of FGF-9 on BFCN or AHCN was somewhat diminished after coupling with polyamine. Experiments to establish optimal condition for FGF-9-polyamine coupling are now underway.

我们检测了成纤维细胞生长因子 (FGF)-9 对胚胎第 17 天获得的大鼠基底前脑胆碱能神经元 (BFCN) 和胚胎第 14 天获得的前角胆碱能神经元 (AHCN) 原代培养物的影响。FGF-9 增强了 AChE 阳性神经元的存活率，增加了其平均胞体大小，并上调了其胆碱能神经元的存活率。 BFCN 和 AHCN 中的乙酰转移酶 (ChAT) 活性。对于 BFCN，FGF-9 的 ChAT 促进作用大约与神经生长因子 (NGF) 一样有效，并且大于碱性成纤维细胞生长因子 (bFGF)、睫状神经营养因子 (CNTF) 或胶质源性神经营养因子 (GDNF) 的作用。同时添加 FGF-9 和 NGF 诱导了极高的 ChAT 水平，表明 FGF-9 和 NGF 可能通过可协同作用的不同途径增强 BFCN 中的胆碱能特性。在培养细胞和成年大鼠脑切片的免疫细胞化学和原位杂交研究中，BFCN 显示 FGF-9 的细胞质免疫染色并表达 FGF-9 信使 RNA；因此，我们得出结论，FGF-9 以自分泌和/或旁分泌方式作用于 BFCN。因此，我们得出结论，FGF-9 可能是阿尔茨海默病和肌萎缩侧索硬化症治疗试验的有希望的候选者。下一步是开发有效的方法将外源性 FGF-9 输送到中枢神经系统。我们将FGF-9与一些多胺偶联，这种FGF-9-多胺复合物可以通过体外BBB系统；然而，FGF-9 对 BFCN 或 AHCN 的神经营养作用在与多胺偶联后有所减弱。目前正在进行实验以确定 FGF-9-多胺偶联的最佳条件。

项目成果

Iwasaki T,Kanda T,Mizusawa H: "Effects of pericytes and various cytokines on integrity of endoneurial monolayer originated from blood-nerve barrier : an in vitro study"Journal of Medical and Dental Sciences. 46(1). 31-40 (1999)

Iwasaki T、Kanda T、Mizusawa H：“周细胞和各种细胞因子对源自血神经屏障的神经内膜单层完整性的影响：一项体外研究”医学和牙科科学杂志。

Kanda T, Iwasaki T, Nakamura S, Kurokawa T, Ikeda K, Mizusawa H: Brain Research. 876. 22-30 (2000)

Kanda T、Iwasaki T、Nakamura S、Kurokawa T、Ikeda K、Mizusawa H：大脑研究。

Kanda T: "Pathology of unmyelinated nerve fibers : a review."Histology and Histopathology. 15. 313-324 (2000)

Kanda T：“无髓神经纤维的病理学：综述。”组织学和组织病理学。

Kanda T, Iwasaki T, Yamawaki M, Mizusawa H, Tai T: "Anti-CM1 antibody facilitates leakage in an in vitro blood-nerve barrier model."Neurology. 55. 585-587 (2000)

Kanda T、Iwasaki T、Yamawaki M、Mizusawa H、Tai T：“抗 CM1 抗体促进体外血神经屏障模型中的渗漏。”神经学。

岩崎孝之,神田隆,水澤英洋: "In vitroモデルをもちいた血液脳関門及び血液神経関門の研究."脳の科学. 21. 553-556 (1999)

Takayuki Iwasaki、Takashi Kanda、Hidehiro Mizusawa：“使用体外模型研究血脑屏障和血神经屏障。” 21. 553-556 (1999)