IBCN   20355
INSTITUTO DE BIOLOGIA CELULAR Y NEUROCIENCIA "PROFESOR EDUARDO DE ROBERTIS"
Unidad Ejecutora - UE
congresos y reuniones científicas
Título:
EFFECTS OF ETHANOL IN PRIMARY CORTICAL NEURONS
Autor/es:
GUADAGNOLI T,ARONNE MP, SAEZ TMM, CALTANA L,POZO DEVOTO V, GIRONACCI M BRUSCO HA.
Lugar:
Rio de Janeiro, Brazil
Reunión:
Simposio; 13th International Symposium on Tobacco Treatment 9th International Symposium on Alcohol and Other Drugs / 1st Latin American; 2010
Institución organizadora:
International Drug Abuse Research Society
Resumen:
The high susceptibility of developing brain neurons to the damaging effects of etanol has been demonstrated using animal and in vitro models of fetal alcohol exposure. Our previous "in vivo" results showed neuronal damage in EtOH-exposed rats. Ethanol (EtOH) is a diffusible molecule with access to the Central Nervous System and it directly acts on neurons. In this work, we have studied the “in vitro” response of primary cortical neurons to EtOH exposure.
Experiments were performed on primary cultures from cortical neurons of Wistar rat at P1 to study the effects of EtOH in morphology and survival in a short-term bioassay. Cells were plated at 3.5 x 106 cells/well in 35mm Nunclon A dishes containing Neurobasal supplemented with B-27 and 2 mM L-glutamine at 37°C in an atmosphere of 95% air and 5% CO2 during 6 days. 12 h before treatment the cell medium was replaced by Neurobasal, and exposed to 25mM, 50mM or 100mM EtOH during 24 h. Control cells were incubated with Neurobasal only. Apoptosis and necrosis were identified using a vital fluorescence rapid test with staining with acridine orange and ethidium bromide and TUNEL staining for apoptosis. MAP-2 immunostaining was used as a specific dendrite marker.We observed an increased of the apoptotic and necrotic neurons percentage in the 50 mM and 100mM EtOH treatment. We observed an increased of TUNEL+ neurons at 50 mM and 100 mM, in concordance to the acridine orange and ethidium bromide stain.We quantified the number of MAP-2+ neurons, the primary dendrite length, secondary dendrite length and number of primary and secondary dendrites. We observed a significantly decrease of the MAP-2 immunoreactivity with 50mM or 100mM EtOH respect to the control treatment.The morphology analysis revealed that cortical neurons treated with 50mM and 100mM EtOH presented a reduction in primary dendrite length and increased in the number of primary dendrite. Our result suggested that exposure to 50mM and 100mM ethanol during 24 hours promotes neuronal death as well as morphological changes in the elongation and branching of dendrites.