title: The molecular basis of the specificity and cross-reactivity of the NeuN epitope of the neuron-specific splicing regulator, Rbfox3.
abstract: Ever since its description and the generation of its defining antibody some 20 years ago, NeuN (Neural Nuclei) has been an invaluable tool for developmental neuroscientist sand neuropathologists to identify neurons and follow their normal or malignant development [corrected].The recent identification of the splicing factor Rbfox3 as the molecule constituting the genuine NeuN epitope has opened up a novel perspective on NeuN immunostaining and its interpretation. Here, we briefly review these recent developments, and we provide a series of data that allow to rationalize the specificity of the NeuN/A60 antibody on aldehyde-fixed tissues on the one hand, and its cross-reactivity with Synapsin I and R3hdm2 on Western blots on the other. We argue that rather than being considered as a mere marker for mature neurons, Rbfox3-mediated NeuN/A60 immunoreactivity may provide a window onto neuronal biology. Specifically, we hypothesize that the phosphorylation-dependent antigenicity of the Rbfox3/NeuN epitope should allow to visualize neuronal physiology realized through Rbfox3, including splicing, on the single-cell level.
authors.affiliation: Department of Molecular and Cellular Physiology, Stanford School of Medicine, Stanford, CA, 94305, USA.
authors.forename: Stephan
authors.initials: S
authors.lastname: Maxeiner
authors.forename: Alexander
authors.initials: A
authors.lastname: Glassmann
authors.forename: Hung-Teh
authors.initials: HT
authors.lastname: Kao
authors.forename: Karl
authors.initials: K
authors.lastname: Schilling
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000918:Antibody Specificity; D001921:Brain / Q000276:immunology; D002478:Cells, Cultured; D003429:Cross Reactions / Q000276:immunology; D004268:DNA-Binding Proteins; D000939:Epitopes / Q000276:immunology; D005260:Female; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009419:Nerve Tissue Proteins / Q000276:immunology* / Q000493:pharmacokinetics*; D009474:Neurons / Q000276:immunology; D009687:Nuclear Proteins / Q000276:immunology* / Q000493:pharmacokinetics*; D010766:Phosphorylation; D016415:Sequence Alignment; D016704:Synapsins / Q000235:genetics / Q000276:immunology*
chemical_list: D004268:DNA-Binding Proteins; D000939:Epitopes; D009419:Nerve Tissue Proteins; C492380:NeuN protein, mouse; D009687:Nuclear Proteins; D016704:Synapsins