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Evaluation of the neurotoxic effects of ethanol on the cerebellar and cortical neurospheres isolated from E14 mouse embryo

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Author(s): Nilesh Kumar Mitra | Kanakeswary Krishnan | Chong Chung Hiong | Yen Nee Ding | Hsiao Lung Eddie | Archana Sikarwar

Journal: Journal of Clinical and Experimental Investigations
ISSN 1309-8578

Volume: 3;
Issue: 4;
Start page: 443;
Date: 2012;
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Keywords: Neural stem cell | alcohol ethyl | in vitro toxicity

ABSTRACT
Objectives: Ethanol exposure during pregnancy has toxiceffects on the neural stem cells of the developing brain.Differential effects of the alcohol on the neural precursorcells from different regions of the brain have not been established.This study was planned to find out the differencein the morphology of the neurosphere (NS) from theembryonic mouse cortex and cerebellum when exposedto ethanol. The dose of ethanol was comparable to bloodalcohol concentration in binge drinking.Materials and methods: The germinal tissues aroundthe lateral ventricle and fourth ventricle of the embryo ofpregnant albino mice (E14) were dissected out. Primarycultures were plated using adequate density of cells inmedia hormone mix (MHM) and growth factors. After 48hours, ethanol was added to the cultures (low dose = 80mg/dl; high dose = 400 mg/dl). NS from different groupswere analyzed using phase-contrast microscopy, Trypanblue exclusion assay and immunostaining with 4’-6-Diamidino-2-phenylindol (DAPI).Results: With low dose ethanol treatment, the mean areaof cortical neurospheres was reduced by 6.9% whereasthat of the cerebellar neurospheres was reduced by51.2%. The mean count of pyknotic cells increased significantly(p
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