A TRIDIMENSIONAL VIEW OF THE ORGANIZATION OF ACTIN FILAMENTS IN CENTRAL NERVOUS SYSTEM USING FLUORESCENT PHOTOOXIDATION

CAPANI F, SARACENO E, BOTI V, AON L, FERNÁNDEZ J, GATO F, KRUSE MS, GIRALDEZ L, ELLISMAN M, COIRINI H

BIOCELL

INCA

Lugar: Mendoza; Año: 2007

0327-9545

Cellular and subcellular organization and distribution of actin filaments have been studied using different type of techniques.  The use of fluorescence photo-oxidation combined with phalloidin conjugates with eosin has allowed examining the precise cellular and subcellular localization of F-actin. Correlative fluorescence light microscopy and transmission electron microscopy studies of F-actin distribution are made relatively easy using this method by morphological and physiological studies. Because phalloidin-eosin is smaller than other markers, this method allows analysing the three-dimensional localization of F-actin using high-resolution light microscopy, three-d serial sections reconstructions and electron tomography.  The combination of this selective staining and 3-D reconstructions provide a valuable tool for revealing aspects of synaptic morphology that are not available using conventional electron microscopy.  By applying this selective staining technique and three-dimensional (3D) imaging we uncovered structural organization of actin in the postsynaptic densities in physiological and pathological conditions.