Open Access

REVIEW

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

by FRANCISCO CAPANI, EZEQUIEL SARACENO, VALERIA ROMINA BOTI, LAURA AON-BERTOLINO, JUAN CARLOS FERNÁNDEZ, FERNANDO GATO, MARIA SOL KRAUSE, LISANDRO GIRALDEZ, MARK H. ELLISMAN, HÉCTOR COIRINI

1. Dept. Bioquímica Humana, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 - 5° P, (1121) Buenos Aires, Argentina.
2. Instituto de Biología y Medicina Experimental, Vuelta de Obligado 2490, (1428) Buenos Aires, Argentina.
3. Laboratorio de Neuroquímica e Biología Celular, Instituto de Ciencias da Saúde, Universidad Federal da Bahia, Bahia, Brasil.
4. National Center for Electron Microscopy and Imaging Research, University of California, San Diego, 9500 Gilman Drive, La Jolla, CA 92093-0608, USA.
Address correspondence to: Francisco Capani, MD, PhD. Departamento de Química Humana, Facultad de Medicina, Universidad de Buenos Aires, Paraguay 2155 - 3ºP, (1121) Buenos Aires, ARGENTINA. E-mail: fcapani@fmed.uba.ar

BIOCELL 2008, 32(1), 1-8. https://doi.org/10.32604/biocell.2008.32.001

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.

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