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

为了研究轴浆转运，将这些大分子注入原代培养的神经细胞，并与含有大分子的人红细胞膜融合。用光血凝素E(PHA-E)和聚乙二醇1000将FITC-BSA或FITC-Ig G固定到培养的小鼠背根神经节(DRG)细胞中。注射效率约为30%，80%的细胞延伸出神经细胞样突起。乳胶珠在轴突中顺行和逆行输送，其顺行速度为0.53微米/秒，逆行速度为0.43微米/秒。速度取决于珠子的大小。从小鼠颈上神经节分离细胞，在盖玻片上培养3d。用视频增强显微镜观察颗粒在神经突起中的运动和神经元的构筑。神经突起中颗粒的运动变得缓慢，最后随着乙酰胆碱的应用而停止。通过去除乙酰胆碱，这些变化是完全可逆的。槟榔碱像乙酰胆碱一样阻止了轴浆运输。QNX可抑制乙酰胆碱的作用。这些药理数据表明，乙酰胆碱的作用与M受体有关。胰岛素激活蛋白(IAP)也可抑制乙酰胆碱对轴浆转运的影响。当轴浆运输停止时，运输的物质从神经突起的微管中脱落。然后，从该区域长出新的轴突分支。这一结果表明，神经递质作为一种可塑性，除了在神经传递中的经典作用外，还可以在调节神经元结构方面发挥重要作用。

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.