LORENZO ALFREDO GUILLERMO
Congresos y reuniones científicas
Título:
Desarrollo de vectores virales para inducir la expresión de la proteína Precursora de Amiloide β (APP) en cultivos celulares
Autor/es:
GHIBAUDO FLORENCIA; HEREDIA FLORENCIA; LORENZO ALFREDO; HELGUERA PABLO
Reunión:
Congreso; XXVII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencia; 2012
Institución organizadora:
Sociedad Argentina de Investigación en Neurociencia (SAN)
Resumen:
B-Amyloid (BA), a peptide generated by the metabolism of B-Amyloid Precursor
Protein (APP), is involved in neurotoxicity in Alzheimer’s Disease (AD).
Understanding APP and its relationship with AB-induced neurotoxicity is critical
to the study of AD. This study was proposed to set parameters in the use of
lentiviruses as a tool for expression of exogenous APP in neurons and in other
cell types. Two kind of lentivirus were generated in the laboratory: One
containing Yellow Fluorescent Protein sequence as a reporter, which was used
to set parameters for the generation of infenctive viral particles, and the other
was intended to express exogenous APP. Both type of viruses were positively
used to infect HEK293T, CHO cells, astrocytes and neurons from rat
hippocampus. The infection efficiency was calculated by epifluorescence
microscopy. The infection percentage was proportional to volume of virus used
to infect HEK293T and CHO cells but not to infect neurons. Although glial cells
were infected, virus showed a greater preference for neurons than glia. We
could express exogenous APP in HEK293T cells and in mixed cultures of
neurons and astrocytes from rat hippocampus. Exogenous APP of infected
CHO cells could not be visualized by Western blot. This technique has proven
to be effective in vitro, and will be adjusted for studies in vivo.
Protein (APP), is involved in neurotoxicity in Alzheimer’s Disease (AD).
Understanding APP and its relationship with AB-induced neurotoxicity is critical
to the study of AD. This study was proposed to set parameters in the use of
lentiviruses as a tool for expression of exogenous APP in neurons and in other
cell types. Two kind of lentivirus were generated in the laboratory: One
containing Yellow Fluorescent Protein sequence as a reporter, which was used
to set parameters for the generation of infenctive viral particles, and the other
was intended to express exogenous APP. Both type of viruses were positively
used to infect HEK293T, CHO cells, astrocytes and neurons from rat
hippocampus. The infection efficiency was calculated by epifluorescence
microscopy. The infection percentage was proportional to volume of virus used
to infect HEK293T and CHO cells but not to infect neurons. Although glial cells
were infected, virus showed a greater preference for neurons than glia. We
could express exogenous APP in HEK293T cells and in mixed cultures of
neurons and astrocytes from rat hippocampus. Exogenous APP of infected
CHO cells could not be visualized by Western blot. This technique has proven
to be effective in vitro, and will be adjusted for studies in vivo.
