A tridimensional view of the organization of actin filaments in the central nervous system by use of fluorescent photooxidation.

CAPANI F, SARACENO E, BOTI VR, AON-BERTOLINO L, FERNÁNDEZ JC, GATO F, KRUSE MS, GIRALDEZ L, ELLISMAN MH, COIRINI H.

BIOCELL

INST HISTOL EMBRIOL-CONICET

Año: 2008 vol. 32 p. 1 - 8

0327-9545

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.