1990 Fiscal Year Final Research Report Summary
The Studied of Axoplasmic Transport Using Cultured Neuronal Cells by Fusion with Erythrocyte Ghosts.
| Project/Area Number |
63480099
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
General physiology
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| Research Institution | Yokohama City University |
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Principal Investigator |
TAKENAKA Toshifumi Yokohama, City Univ., Physiology, Prof.,, 医学部, 教授 (00045999)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAKAMI Tadashi Yokohama, City Univ., Physiology, Lecture., 医学部, 助手 (60177649)
HORI Hideaki Yokohama, City Univ., Physiology, Lecture., 医学部, 助手 (30117759)
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| Project Period (FY) |
1988 – 1990
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| Keywords | axoplamic transport / cell fusion / erythocyte ghosts / cultured cell / dorsal root ganlion / supper cervical ganglion / latex beads / axonal flow |
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| Research Abstract |
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.
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[Publications] Yamaguchi, M., Yamada, S., Daimon, N., Yamamoto, I., Kawakami, T. & Takenaka, T.: "Electromagnetic mechanism of magnetic nerve stimulation." J. Appl. Phys.66. 1459-1465 (1989)
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[Publications] Kawakami, T., Hikawa, N., Kanou, M., Gotoh, H., Takenaka, T. & Horie, H.: "Axoplasmic transport of mitochondria in tissue cultured neuron." Jpn. J. Physiol. 39(Suppl). S45 (1989)
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