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KMID : 1102020180480030055
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Applied Microscopy
2018 Volume.48 No. 3 p.55 ~ p.61
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Role of Actin Filament on Synaptic Vesicle Pooling in Cultured Hippocampal Neuron
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Lee Se-Jeong
Kim Hyun-Wook
Na Ji-Eun
Kim Da-Som
Kim Dai-Hyun
Ryu Jae-Ryun
Sun Woong
Rhyu Im-Joo
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Abstract
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The synaptic vesicle is a specialized structure in presynaptic terminals that stores various neurotransmitters. The actin filament has been proposed for playing an important role in mobilizing synaptic vesicles. To understand the role of actin filament on synaptic vesicle pooling, we characterized synaptic vesicles and actin filament after treatment of brain-derived neurotrophic factor (BDNF) or Latrunculin A on primary cultured neuron from rat embryo hippocampus. Western blots revealed that BDNF treatment increased the expression of synapsin I protein, but Latrunculin A treatment decreased the synapsin I protein expression. The increased expression of synapsin I after BDNF disappeared by the treatment of Latrunculin A. Three-dimensional (3D) tomography of synapse showed that more synaptic vesicles localized near the active zone and total number of synaptic vesicles increased after treatment of BDNF. But the number of synaptic vesicle was 2.5-fold reduced in presynaptic terminals and the loss of filamentous network was observed after Latrunculin A application. The treatment of Latruculin A after preincubation of BDNF group showed that synaptic vesicle number was similar to that of control group, but filamentous structures were not restored. These data suggest that the actin filament plays a significant role in synaptic vesicles pooling in presynaptic terminals.
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KEYWORD
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Synaptic vesicle, Actin filament, Brain-derived neurotrophic factor, Latrunculin A, Electron tomography
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