A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.
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.
Capani
Francisco
F
Dept. Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 - 5o P, (1121) Buenos Aires, Argentina. fcapani@fmed.uba.ar
Saraceno
Ezequiel
E
Boti
Valeria Romina
VR
Aon-Bertolino
Laura
L
Fernández
Juan Carlos
JC
Gato
Fernándo
F
Kruse
Maria Sol
MS
Krause
Maria Sol
MS
Giraldez
Lisandro
L
Ellisman
Mark H
MH
Coirini
Héctor
H
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
D000199:Actins; D005456:Fluorescent Dyes; D010590:Phalloidine; D004801:Eosine Yellowish-(YS)