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Neuron-glia interaction - novel neural trophic factors and their molecular action mechanism -

神经元-胶质细胞相互作用-新型神经营养因子及其分子作用机制-

基本信息

批准号：
02454158
负责人：
KATO Taiji
金额：
$ 4.03万
依托单位：
Nagoya City University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

Two kinds of novel neural trophic factors were detected in von Recklinghausen neurofibroma (NF1) extracts. one of two, neuroblastoma growth factor (Mr<5 kDa), promotes the proliferation of human neuroblastoma cell and survival and neurite-extension of rat cortical neuron, but is distinct from nerve growth factor (NGF) or NGF-like factors. The other trophic factor is a glial growth inhibitor (Mr=100 k), which suppresses the growth of glioma cell lines, astrocytoma, glioblastoma, oligodendroglioma and Schwannoma. These factors do not appear to be previously identified cytokines or growth factors such as interleukins, granulocyte colony-stimu-lating factor, NGF and fibroblast growth factor, Ciliary neurotrophic factor-like activity was also detected in the extracts. Anovel endogenous polypeptide inhibitor of proliferation and DNA synthesis of glial cells, gliostatin, was purified from the extracts of neurofibroma by a procedure comprising column chromatography with Blue-Toyopearl and DEAE-Sephacel, and high-performance liquid chromatography with Butyl-Toyopearl, Hydroxylapatite and Mono Q. monoclonal antibody raised against partially purified (Hydroxylapatite step) gliostatin adsorbed the growth inhibitory activity of gliostatin and immunochemically visualized the putative gliostatin bands on Western blot analyses, but showed no cross-reactivity with known cytokinesis. Although the product showed an apparent Mr=100 k accompanied by an inhibitory activity on Superdex 200 column chromatography, it migrated at a lower apparent Mr=50 k under the reducing condition on Western blotting, indicating that a homodimeric structure of native gliostatin consisted of 50 KDa subcomponents. Gliostatin was a potent growth inhibitor acting at nM concentration against all glial tumor cells d glia maturation factor (GMF)-stimulated astroblasts but not neuronal cells.

从von Reckinghausen神经纤维瘤(NF1)提取物中检测到两种新的神经营养因子。其中神经母细胞瘤生长因子(5 KDa)能促进人神经母细胞瘤细胞的增殖和大鼠皮质神经元的存活和突起生长，但不同于神经生长因子(NGF)或NGF样因子。另一种营养因子是胶质生长抑制因子(MR=100k)，它抑制胶质瘤细胞系、星形细胞瘤、胶质母细胞瘤、少突胶质细胞瘤和神经鞘瘤的生长。这些因子似乎不是以前发现的细胞因子或生长因子，如白介素类、粒细胞集落刺激因子、神经生长因子和成纤维细胞生长因子，在提取物中还检测到睫状神经营养因子样活性。一种新型的内源性神经胶质细胞增殖和DNA合成的多肽抑制物--胶质抑素是从神经纤维瘤的提取物中提纯的，该方法包括用Blue-Toyota opearl和DEAE-Sephacel柱层析，用丁基-toyopearl，羟基磷灰石和Mono-Q的高效液相层析。针对部分纯化的(羟基磷灰石步骤)gliostatin的单抗吸附了gliostatin的生长抑制活性，并在Western印迹分析中显示了可能的gliostatin条带，但与已知的细胞质分裂没有交叉反应。虽然该产物在Superdex 200柱层析上表现出明显的MR=100k的抑制活性，但在Western blotting的还原条件下，它在较低的表观MR=50k处迁移，表明天然的胶质抑制素是由50 KDa的亚组分组成的同源二聚体结构。Gliostatin是一种有效的生长抑制物，在NM浓度下对所有胶质瘤细胞和胶质成熟因子(GMF)刺激的星形胶质细胞起作用，但对神经细胞没有作用。