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Measurement of saturation processes in glutamatergic and GABAergic synapse densities during long-term development of cultured rat cortical networks

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Title: Measurement of saturation processes in glutamatergic and GABAergic synapse densities during long-term development of cultured rat cortical networks
Authors: Ito, Daisuke Browse this author
Komatsu, Takumi Browse this author
Gohara, Kazutoshi Browse this author →KAKEN DB
Keywords: Dissociated culture
Immunofluorescence staining
Multi-electrode array
Issue Date: 9-Oct-2013
Publisher: Elsevier science bv
Journal Title: Brain research
Volume: 1534
Start Page: 22
End Page: 32
Publisher DOI: 10.1016/j.brainres.2013.08.004
PMID: 23948099
Abstract: The aim of this study was to clarify the saturation processes of excitatory and inhibitory synapse densities during the long-term development of cultured neuronal networks. For this purpose, we performed a long-term culture of rat cortical cells for 35 days in vitro (DIV). During this culture period, we labeled glutamatergic and GABAergic synapses separately using antibodies against vesicular glutamate transporter 1 (VGluT1) and vesicular transporter of gamma-aminobutyric acid (VGAT). The densities and distributions of both types of synaptic terminals were measured simultaneously. Observations and subsequent measurements of immunofluorescence demonstrated that the densities of both types of antibody-labeled terminals increased gradually from 7 to 21-28 DIV. The densities did not show a further increase at 35 DIV and tended to become saturated. Triple staining with VGluT1, VGAT, and microtubule-associated protein 2 (MAP2) enabled analysis of the distribution of both types of synapses, and revealed that the densities of the two types of synaptic terminals on somata were not significantly different but that glutamatergic synapses predominated on the dendrites during long-term culture. However, some neurons did not fall within this distribution, suggesting differences in synapse distribution on target neurons. The electrical activity also showed an initial increase and subsequent saturation of the firing rate and synchronized burst rate during long-term culture, and the number of days of culture to saturation from the initial increase followed the same pattern under this culture condition. (C) 2013 Elsevier B.V. All rights reserved.
Type: article (author version)
Appears in Collections:工学院・工学研究院 (Graduate School of Engineering / Faculty of Engineering) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 郷原 一寿

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