title: Deconstructing complexity: serial block-face electron microscopic analysis of the hippocampal mossy fiber synapse.
abstract: The hippocampal mossy fiber (MF) terminal is among the largest and most complex synaptic structures in the brain. Our understanding of the development of this morphologically elaborate structure has been limited because of the inability of standard electron microscopy techniques to quickly and accurately reconstruct large volumes of neuropil. Here we use serial block-face electron microscopy (SBEM) to surmount these limitations and investigate the establishment of MF connectivity during mouse postnatal development. Based on volume reconstructions, we find that MF axons initially form bouton-like specializations directly onto dendritic shafts, that dendritic protrusions primarily arise independently of bouton contact sites, and that a dramatic increase in presynaptic and postsynaptic complexity follows the association of MF boutons with CA3 dendritic protrusions. We also identify a transient period of MF bouton filopodial exploration, followed by refinement of sites of synaptic connectivity. These observations enhance our understanding of the development of this highly specialized synapse and illustrate the power of SBEM to resolve details of developing microcircuits at a level not easily attainable with conventional approaches.
authors.affiliation: Neurobiology Section, Division of Biological Sciences and National Center for Microscopy and Imaging Research, University of California, San Diego, La Jolla, California 92093, USA.
authors.forename: Scott A
authors.initials: SA
authors.lastname: Wilke
authors.forename: Joseph K
authors.initials: JK
authors.lastname: Antonios
authors.forename: Eric A
authors.initials: EA
authors.lastname: Bushong
authors.forename: Ali
authors.initials: A
authors.lastname: Badkoobehi
authors.forename: Elmar
authors.initials: E
authors.lastname: Malek
authors.forename: Minju
authors.initials: M
authors.lastname: Hwang
authors.forename: Masako
authors.initials: M
authors.lastname: Terada
authors.forename: Mark H
authors.initials: MH
authors.lastname: Ellisman
authors.forename: Anirvan
authors.initials: A
authors.lastname: Ghosh
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D001369:Axons / Q000648:ultrastructure; D003712:Dendrites / Q000648:ultrastructure; D007091:Image Processing, Computer-Assisted; D051379:Mice; D008810:Mice, Inbred C57BL; D008854:Microscopy, Electron / Q000379:methods*; D019599:Mossy Fibers, Hippocampal / Q000648:ultrastructure*; D009412:Nerve Fibers / Q000648:ultrastructure*; D019581:Neuropil / Q000648:ultrastructure; D017729:Presynaptic Terminals / Q000648:ultrastructure; D011554:Pseudopodia / Q000648:ultrastructure; D011786:Quality Control; D012984:Software; D013569:Synapses / Q000648:ultrastructure*