title: Mapping the connectivity of serotonin transporter immunoreactive axons to excitatory and inhibitory neurochemical synapses in the mouse limbic brain.
abstract: Serotonin neurons arise from the brainstem raphe nuclei and send their projections throughout the brain to release 5-HT which acts as a modulator of several neuronal populations. Previous electron microscopy studies in rats have morphologically determined the distribution of 5-HT release sites (boutons) in certain brain regions and have shown that 5-HT containing boutons form synaptic contacts that are either symmetric or asymmetric. In addition, 5-HT boutons can form synaptic triads with the pre- and postsynaptic specializations of either symmetrical or asymmetrical synapses. However, due to the labor intensive processing of serial sections required by electron microscopy, little is known about the neurochemical properties or the quantitative distribution of 5-HT triads within whole brain or discrete subregions. Therefore, we used a semi-automated approach that combines immunohistochemistry and high-resolution confocal microscopy to label serotonin transporter (SERT) immunoreactive axons and reconstruct in 3D their distribution within limbic brain regions. We also used antibodies against key pre- (synaptophysin) and postsynaptic components of excitatory (PSD95) or inhibitory (gephyrin) synapses to (1) identify putative 5-HTergic boutons within SERT immunoreactive axons and, (2) quantify their close apposition to neurochemical excitatory or inhibitory synapses. We provide a 5-HTergic axon density map and have determined the ratio of synaptic triads consisting of a 5-HT bouton in close proximity to either neurochemical excitatory or inhibitory synapses within different limbic brain areas. The ability to model and map changes in 5-HTergic axonal density and the formation of triadic connectivity within whole brain regions using this rapid and quantitative approach offers new possibilities for studying neuroplastic changes in the 5-HTergic pathway.
authors.affiliation: Translational Research Institute, Queensland University of Technology, Brisbane, Qld 4059, Australia.
authors.forename: Arnauld
authors.initials: A
authors.lastname: Belmer
authors.affiliation: Translational Research Institute, Queensland University of Technology, Brisbane, Qld 4059, Australia.
authors.forename: Paul M
authors.initials: PM
authors.lastname: Klenowski
authors.affiliation: Translational Research Institute, Queensland University of Technology, Brisbane, Qld 4059, Australia.
authors.forename: Omkar L
authors.initials: OL
authors.lastname: Patkar
authors.affiliation: Translational Research Institute, Queensland University of Technology, Brisbane, Qld 4059, Australia. selena.bartlett@qut.edu.au.|Institute of Health and Biomedical Innovation (IHBI), Queensland University of Technology, Brisbane, Australia. selena.bartlett@qut.edu.au.
authors.forename: Selena E
authors.initials: SE
authors.lastname: Bartlett
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001369:Axons / Q000378:metabolism* / Q000648:ultrastructure; D001931:Brain Mapping; D002352:Carrier Proteins / Q000378:metabolism; D000076122:Disks Large Homolog 4 Protein; D051528:Guanylate Kinases / Q000378:metabolism; D021621:Imaging, Three-Dimensional; D008032:Limbic System / Q000166:cytology* / Q000502:physiology; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008854:Microscopy, Electron; D017729:Presynaptic Terminals / Q000378:metabolism / Q000648:ultrastructure; D012701:Serotonin / Q000378:metabolism; D050486:Serotonin Plasma Membrane Transport Proteins / Q000378:metabolism*; D013569:Synapses / Q000378:metabolism* / Q000502:physiology* / Q000648:ultrastructure; D016708:Synaptophysin / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; D000076122:Disks Large Homolog 4 Protein; C501447:Dlg4 protein, mouse; D008565:Membrane Proteins; D050486:Serotonin Plasma Membrane Transport Proteins; D016708:Synaptophysin; C073441:gephyrin; D012701:Serotonin; D051528:Guanylate Kinases
keywords: 3D reconstruction; Gephyrin; Neuromodulation; PSD-95; Serotonin; Synaptophysin