Abstract

The study of the mechanism of neuronal cell death has been an important aspect of cell biology in the last decade. Considering the somewhat non-specific nature of the methods and the complexity of the neuronal cell cycle, a multi-technique approach has been suggested. This study is aimed at describing a method for demonstration of neuronal degeneration in the visual cortex (molecular layer) of adult wistar rats. It involves the use of colloidal gold (40 nm), conjugated with an antibody against protein G (transmembrane protein) to generate fluorescence; in essence to label the regions of cells with functional membrane, while the region of necrotic cells are empty. This method is complemented by histology to generate overlap images and to detect loss of membrane integrity in neurons at the early stages of progressive neuronal degeneration. An in vivo approach was used; 50 mg/Kg BW of potassium cyanide-Sigma (KCN) was administered orally to adult Wistar rats to induce neurotoxicity. The animals were sacrificed, and sections were labeled with anti Protein G-colloidal gold conjugate. The 40 nm gold particles generated fluorescence in the region of the cells with functional membrane properties, while empty spaces were found in regions of necrotic cells at a magnification of 400× (low magnification).

Key words: Immunoglobin G (IgG), nanoparticles, cell death, bovine serum albumin (BSA).