title: Electron cryotomography of postsynaptic densities during development reveals a mechanism of assembly.
abstract: Postsynaptic densities (PSDs) are responsible for organizing receptors and signaling proteins that regulate excitatory transmission in the mammalian brain. To better understand the assembly and 3D organization of this synaptic structure, we employed electron cryotomography to visualize general and fine structural details of PSDs isolated from P2, P14, P21 and adult forebrain in the absence of fixatives and stains. PSDs at P2 are a loose mesh of filamentous and globular proteins and during development additional protein complexes are recruited onto the mesh. Quantitative analysis reveals that while the surface area of PSDs is relatively constant, the thickness and protein occupancy of the PSD volume increase dramatically between P14 and adult. One striking morphological feature is the appearance of lipid raft-like structures, first evident in PSDs from 14 day old animals. These detergent-resistant membranes stain for GM1 ganglioside and their terminations can be clearly seen embedded in protein "bowls" within the PSD complex. In total, these results lead to the conclusion that the PSD is assembled by the gradual recruitment and stabilization of proteins within an initial mesh that systematically adds complexity to the structure.
authors.affiliation: Department of Neurobiology and Anatomy, University of Texas Medical School at Houston, Houston, TX 77030, USA.
authors.forename: M T
authors.initials: MT
authors.lastname: Swulius
authors.forename: M M
authors.initials: MM
authors.lastname: Farley
authors.forename: M A
authors.initials: MA
authors.lastname: Bryant
authors.forename: M N
authors.initials: MN
authors.lastname: Waxham
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D055032:Electron Microscope Tomography / Q000379:methods*; D021621:Imaging, Three-Dimensional / Q000379:methods; D055495:Neurogenesis / Q000502:physiology*; D057907:Post-Synaptic Density / Q000502:physiology* / Q000648:ultrastructure*; D016548:Prosencephalon / Q000254:growth & development* / Q000648:ultrastructure*; D051381:Rats