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The Studied of Axoplasmic Transport Using Cultured Neuronal Cells by Fusion with Erythrocyte Ghosts.

通过与红细胞幽灵融合来研究使用培养的神经元细胞的轴浆运输。

基本信息

批准号：
63480099
负责人：
TAKENAKA Toshifumi
金额：
$ 4.1万
依托单位：
Yokohama 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：
1988
资助国家：
日本
起止时间：
1988 至 1990
项目状态：
已结题

项目摘要

In order to study the axoplasmic transport, the macromolecules were injected into primary cultured neuronal cells by fusion with human erythrocyte ghosts containing macromolecules. FITC-BSA or FITC-IgG trapped into cultured mouse dorsal root ganglion (DRG) cells using photohaemaagglutinin E (PHA-E) and polyethylene glycol 1000. The injection efficiency was about 30% and 80% of the cells extended neurites like neuronal cells. The latex beads transported in neurites both anterogradely and retrogradely The velocities of beads were 0.53 mu m/sec in anterograde direction and 0.43 mu m/sec in retrograde direction respectively. The velocity depends on the size of beads. The velocity of small beads is faster than that of bigger beads.Cells were dissociated from the superior cervical ganglion (SCG) of mice and cultured on coverslips for 3 days. The particle movements in neurites and neuronal architecture were observed by video-enhanced microscopy. The movements of particles in neurites becomes slow and finally stops with the application of Acetylcholine. These changes were completely reversible by removing Acetylcholine. Arecoline stopped the axoplasmic transport like acetylcholine. QNX restrained the acetylcholine effects. These pharmacological data show that the effect of acetylcholine related with muscarinic receptor. Inslet Activating Protein (IAP) also restrained the effect of acetylcholine on the axoplasmic transport. When axoplasmic transport was stopped, the transported material dropped off from the microtubule in the neurites. Then from that area the new branches of the neurites grow. This result demonstrates that neurotransmitter can play a prominent role in regulating neuronal architecture as a type of plasticity besides their classical role in neurotransmission.

为了研究轴浆运输，将大分子注射到原代培养的神经细胞中，通过与含有大分子的人红细胞血影融合。使用光血凝素E（PHA-E）和聚乙二醇1000将FITC-BSA或FITC-IgG捕获到培养的小鼠背根神经节（DRG）细胞中。注射效率约为30%，80%的细胞像神经元细胞一样伸出突起。乳胶珠在神经突中的运输既有顺向的，也有逆向的，其速度分别为0.53 μ m/sec和0.43 μ m/sec。速度取决于珠子的大小。从小鼠颈上级神经节（SCG）分离细胞，在盖玻片上培养3天。通过视频增强显微镜观察突起内颗粒的运动和神经元的结构。颗粒在神经突中的运动变得缓慢，并最终停止与应用乙酰胆碱。这些变化通过去除乙酰胆碱完全可逆。槟榔碱像乙酰胆碱一样阻断轴浆运输。QNX抑制乙酰胆碱效应。这些药理学数据表明，乙酰胆碱的作用与M受体有关。胰岛激活蛋白（IAP）也抑制乙酰胆碱对轴浆转运的影响。当轴浆运输停止时，运输的物质从轴突中的微管脱落。然后从那个区域长出新的神经突分支。这一结果表明，神经递质除了在神经传递中发挥经典作用外，还可以作为一种可塑性在调节神经元结构中发挥重要作用。