title: Molecular in situ topology of Aczonin/Piccolo and associated proteins at the mammalian neurotransmitter release site.
abstract: The protein machinery of neurotransmitter exocytosis requires efficient orchestration in space and time, for speed and precision of neurotransmission and also for synaptic ontogeny and plasticity. However, its spatial organization in situ is virtually unknown. Aczonin/Piccolo is a putative organizer protein of mammalian active zones. We determined by immunogold electron microscopy (EM) (i) the spatial arrangement (i.e., topology) of 11 segments of the Aczonin polypeptide in situ, and correlated it to (ii) the positioning of Aczonin-interacting domains of Bassoon, CAST/ELKS, Munc13, and RIM and (iii) the ultrastructurally defined presynaptic macromolecular aggregates known as dense projections and synaptic ribbons. At conventional synapses, Aczonin assumes a compact molecular topology within a layer 35 to 80 nm parallel to the plasma membrane (PM), with a "trunk" sitting on the dense projection top and a C-terminal "arm" extending down toward the PM and sideward to the dense projection periphery. At ribbon synapses, Aczonin occupies the whole ribbon area. Bassoon colocalizes with Aczonin at conventional synapses but not at ribbon synapses. At both conventional and ribbon synapses, CAST, Munc13, and RIM are segregated from Aczonin, closer to the PM, and Aczonin is positioned such that it may control the access of neurotransmitter vesicles to the fusion site.
authors.affiliation: Department of Neuroscience, Uppsala University, S-75124 Uppsala, Sweden.
authors.forename: Christoph
authors.initials: C
authors.lastname: Limbach
authors.forename: Michael M
authors.initials: MM
authors.lastname: Laue
authors.forename: Xiaolu
authors.initials: X
authors.lastname: Wang
authors.forename: Bin
authors.initials: B
authors.lastname: Hu
authors.forename: Nadine
authors.initials: N
authors.lastname: Thiede
authors.forename: Greta
authors.initials: G
authors.lastname: Hultqvist
authors.forename: Manfred W
authors.initials: MW
authors.lastname: Kilimann
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D019204:GTP-Binding Proteins / Q000378:metabolism; D015151:Immunoblotting; D016253:Microscopy, Immunoelectron; D046912:Multiprotein Complexes / Q000378:metabolism / Q000648:ultrastructure; D009419:Nerve Tissue Proteins / Q000378:metabolism; D009479:Neuropeptides / Q000378:metabolism*; D018377:Neurotransmitter Agents / Q000378:metabolism*; D011485:Protein Binding; D051381:Rats; D017207:Rats, Sprague-Dawley; D013569:Synapses / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C113688:Bsn protein, rat; D003598:Cytoskeletal Proteins; C479909:ERC1 protein, rat; D046912:Multiprotein Complexes; D009419:Nerve Tissue Proteins; D009479:Neuropeptides; D018377:Neurotransmitter Agents; C405514:Pclo protein, rat; C107293:Rim protein, mammalian; C103912:Unc13a protein, rat; D019204:GTP-Binding Proteins