title: Sub-synaptic, multiplexed analysis of proteins reveals Fragile X related protein 2 is mislocalized inFmr1KO synapses.
abstract: The distribution of proteins within sub-synaptic compartments is an essential aspect of their neurological function. Current methodologies, such as electron microscopy (EM) and super-resolution imaging techniques, can provide the precise localization of proteins, but are often limited to a small number of one-time observations with narrow spatial and molecular coverage. The diversity of synaptic proteins and synapse types demands synapse analysis on a scale that is prohibitive with current methods. Here, we demonstrate SubSynMAP, a fast, multiplexed sub-synaptic protein analysis method using wide-field data from deconvolution array tomography (ATD). SubSynMAP generates probability distributions for that reveal the functional range of proteins within the averaged synapse of a particular class. This enables the differentiation of closely juxtaposed proteins. Using this method, we analyzed 15 synaptic proteins in normal and Fragile X mental retardation syndrome (FXS) model mouse cortex, and revealed disease-specific modifications of sub-synaptic protein distributions across synapse classes and cortical layers.
authors: Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, United States.|Center for Sleep Sciences and Medicine, Stanford University School of Medicine, Stanford, United States.|Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States.
authors: Gordon X
authors: 0000-0002-2707-7118
authors: GX
authors: Wang
authors: Department of Molecular and Cellular Physiology, Stanford University School of Medicine, Stanford, United States.|Allen Institute for Brain Science, Seattle, USA.
authors: Stephen J
authors: SJ
authors: Smith
authors: Department of Psychiatry and Behavioral Sciences, Stanford University School of Medicine, Stanford, United States.|Center for Sleep Sciences and Medicine, Stanford University School of Medicine, Stanford, United States.|INSERM 1024, Ecole Normale Supérieure, Paris, France.
authors: Philippe
authors: P
authors: Mourrain
mesh_terms: D000818:Animals; D004195:Disease Models, Animal; D051860:Fragile X Messenger Ribonucleoprotein 1 / Q000235:genetics*; D005600:Fragile X Syndrome / Q000473:pathology*; D055786:Gene Knockout Techniques; D051379:Mice; D018345:Mice, Knockout; D061848:Optical Imaging / Q000379:methods*; D016601:RNA-Binding Proteins / Q000032:analysis*; D013569:Synapses / Q000737:chemistry*
chemical_list: D051860:Fragile X Messenger Ribonucleoprotein 1; D016601:RNA-Binding Proteins; C512309:FXR2 protein, mouse; C497684:Fmr1 protein, mouse
keywords: FXR2P; FXS; mouse; neuroscience; synapse; synapse classification