title: A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.
abstract: Cellular and subcellular organization and distribution of actin filaments have been studied with various techniques. The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed the examination of the precise cellular and subcellular location of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are facilitated with this method for morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows the analysis of the three-dimensional location of F-actin with high-resolution light microscopy, three-d serial sections reconstructions, and electron tomography. The combination of selective staining and three-dimensional reconstructions provide a valuable tool for revealing aspects of the synaptic morphology that are not available when conventional electron microscopy is used. By applying this selective staining technique and three-dimensional imaging, we uncovered the structural organization of actin in the postsynaptic densities in physiological and pathological conditions.
authors.affiliation: Dept. Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 - 5o P, (1121) Buenos Aires, Argentina. fcapani@fmed.uba.ar
authors.forename: Francisco
authors.initials: F
authors.lastname: Capani
authors.forename: Ezequiel
authors.initials: E
authors.lastname: Saraceno
authors.forename: Valeria Romina
authors.initials: VR
authors.lastname: Boti
authors.forename: Laura
authors.initials: L
authors.lastname: Aon-Bertolino
authors.forename: Juan Carlos
authors.initials: JC
authors.lastname: Fernández
authors.forename: Fernándo
authors.initials: F
authors.lastname: Gato
authors.forename: Maria Sol
authors.initials: MS
authors.lastname: Kruse
authors.forename: Maria Sol
authors.initials: MS
authors.lastname: Krause
authors.forename: Lisandro
authors.initials: L
authors.lastname: Giraldez
authors.forename: Mark H
authors.initials: MH
authors.lastname: Ellisman
authors.forename: Héctor
authors.initials: H
authors.lastname: Coirini
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism / Q000648:ultrastructure*; D000199:Actins / Q000378:metabolism; D000818:Animals; D002490:Central Nervous System / Q000378:metabolism / Q000648:ultrastructure*; D004801:Eosine Yellowish-(YS) / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000494:pharmacology*; D006801:Humans; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods; D008958:Models, Molecular; D010084:Oxidation-Reduction; D010590:Phalloidine / Q000494:pharmacology; D017785:Photons; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000199:Actins; D005456:Fluorescent Dyes; D010590:Phalloidine; D004801:Eosine Yellowish-(YS)