Desarrollo de vectores virales para inducir la expresión de la proteína Precursora de Amiloide β (APP) en cultivos celulares

GHIBAUDO FLORENCIA; HEREDIA FLORENCIA; LORENZO ALFREDO; HELGUERA PABLO

Congreso; XXVII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencia; 2012

Sociedad Argentina de Investigación en Neurociencia (SAN)

B-Amyloid (BA), a peptide generated by the metabolism of B-Amyloid PrecursorProtein (APP), is involved in neurotoxicity in Alzheimer’s Disease (AD).Understanding APP and its relationship with AB-induced neurotoxicity is criticalto the study of AD. This study was proposed to set parameters in the use oflentiviruses as a tool for expression of exogenous APP in neurons and in othercell types. Two kind of lentivirus were generated in the laboratory: Onecontaining Yellow Fluorescent Protein sequence as a reporter, which was usedto set parameters for the generation of infenctive viral particles, and the otherwas intended to express exogenous APP. Both type of viruses were positivelyused to infect HEK293T, CHO cells, astrocytes and neurons from rathippocampus. The infection efficiency was calculated by epifluorescencemicroscopy. The infection percentage was proportional to volume of virus usedto infect HEK293T and CHO cells but not to infect neurons. Although glial cellswere infected, virus showed a greater preference for neurons than glia. Wecould express exogenous APP in HEK293T cells and in mixed cultures ofneurons and astrocytes from rat hippocampus. Exogenous APP of infectedCHO cells could not be visualized by Western blot. This technique has provento be effective in vitro, and will be adjusted for studies in vivo.