title: Synapse-dependent and independent mechanisms of thalamocortical axon branching are regulated by neuronal activity.
abstract: Axon branching and synapse formation are critical processes for establishing precise circuit connectivity. These processes are tightly regulated by neural activity, but the relationship between them remains largely unclear. We use organotypic coculture preparations to examine the role of synapse formation in the activity-dependent axon branching of thalamocortical (TC) projections. To visualize TC axons and their presynaptic sites, two plasmids encoding DsRed and EGFP-tagged synaptophysin (SYP-EGFP) were cotransfected into a small number of thalamic neurons. Time-lapse imaging of individual TC axons showed that most branches emerged from SYP-EGFP puncta, indicating that synapse formation precedes emergences of axonal branches. We also investigated the effects of neuronal activity on axon branching and synapse formation by manipulating spontaneous firing activity of thalamic cells. An inward rectifying potassium channel, Kir2.1, and a bacterial voltage-gated sodium channel, NaChBac, were used to suppress and promote firing activity, respectively. We found suppressing neural activity reduced both axon branching and synapse formation. In contrast, increasing neural activity promoted only axonal branch formation. Time-lapse imaging of NaChBac-expressing cells further revealed that new branches frequently appeared from the locations other than SYP-EGFP puncta, indicating that enhancing activity promotes axonal branch formation due to an increase of branch emergence at nonsynaptic sites. These results suggest that presynaptic locations are hotspots for branch emergence, and that frequent firing activity can shift branch emergence to a synapse-independent process.
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
authors.forename: Naoyuki
authors.initials: N
authors.lastname: Matsumoto
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
authors.forename: Maki
authors.initials: M
authors.lastname: Hoshiko
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Sugo
authors.affiliation: Division of Brain Structure and Function, Faculty of Medical Science, University of Fukui, Eiheiji, Yoshida, 910-1193, Japan.
authors.forename: Yugo
authors.initials: Y
authors.lastname: Fukazawa
authors.affiliation: Laboratory of Cellular and Molecular Neurobiology Graduate School of Frontier Biosciences, Osaka University, Suita, Osaka, 565-0871, Japan.
authors.forename: Nobuhiko
authors.initials: N
authors.lastname: Yamamoto
mesh_terms: D000818:Animals; D001369:Axons / Q000502:physiology* / Q000648:ultrastructure; D001921:Brain / Q000254:growth & development* / Q000648:ultrastructure; D018274:Electroporation; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009434:Neural Pathways / Q000254:growth & development* / Q000648:ultrastructure; D055495:Neurogenesis / Q000502:physiology*; D009474:Neurons / Q000502:physiology / Q000648:ultrastructure; D009924:Organ Culture Techniques; D018408:Patch-Clamp Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D013569:Synapses / Q000502:physiology* / Q000648:ultrastructure; D059008:Time-Lapse Imaging
keywords: axon guidance; axonal arborization; cortical development; synaptogenesis; thalamus