title: A cell surface biotinylation assay to reveal membrane-associated neuronal cues: Negr1 regulates dendritic arborization.
abstract: A complex and still not comprehensively resolved panel of transmembrane proteins regulates the outgrowth and the subsequent morphological and functional development of neuronal processes. In order to gain a more detailed description of these events at the molecular level, we have developed a cell surface biotinylation assay to isolate, detect, and quantify neuronal membrane proteins. When we applied our assay to investigate neuron maturation in vitro, we identified 439 differentially expressed proteins, including 20 members of the immunoglobulin superfamily. Among these candidates, we focused on Negr1, a poorly described cell adhesion molecule. We demonstrated that Negr1 controls the development of neurite arborization in vitro and in vivo. Given the tight correlation existing among synaptic cell adhesion molecules, neuron maturation, and a number of neurological disorders, our assay results are a useful tool that can be used to support the understanding of the molecular bases of physiological and pathological brain function.
authors.affiliation: Institute of Neuroscience CNR, 20129 Milano, Italy;
authors.forename: Francesca
authors.initials: F
authors.lastname: Pischedda
authors.forename: Joanna
authors.initials: J
authors.lastname: Szczurkowska
authors.forename: Maria Daniela
authors.initials: MD
authors.lastname: Cirnaru
authors.forename: Florian
authors.initials: F
authors.lastname: Giesert
authors.forename: Elena
authors.initials: E
authors.lastname: Vezzoli
authors.forename: Marius
authors.initials: M
authors.lastname: Ueffing
authors.forename: Carlo
authors.initials: C
authors.lastname: Sala
authors.forename: Maura
authors.initials: M
authors.lastname: Francolini
authors.forename: Stefanie M
authors.initials: SM
authors.lastname: Hauck
authors.forename: Laura
authors.initials: L
authors.lastname: Cancedda
authors.forename: Giovanni
authors.initials: G
authors.lastname: Piccoli
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods*; D019921:Biotinylation; D015816:Cell Adhesion Molecules, Neuronal / Q000378:metabolism*; D002454:Cell Differentiation; D002462:Cell Membrane / Q000378:metabolism*; D048430:Cell Shape; D002478:Cells, Cultured; D003712:Dendrites / Q000378:metabolism*; D049229:Dendritic Spines / Q000378:metabolism; D020868:Gene Silencing; D057809:HEK293 Cells; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D055495:Neurogenesis; D013569:Synapses / Q000378:metabolism
chemical_list: D015816:Cell Adhesion Molecules, Neuronal; D008565:Membrane Proteins; C000592528:NEGR1 protein, mouse