Nilesh Kumar MİTRA, Kanakeswary KRISHNAN, Chong Chung HIONG, Yen Nee DING, Hsiao Lung EDDIE, Archana SIKARWAR

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

Amaç: Gebelikte etanol maruziyeti, gelişen beyindekinöral kök hücreler üzerine toksik etkilere sahiptir. Beynin değişik bölgelerindeki nöral öncül hücreler üzerine alkolün farklı etkileri henüz belirlenmemiştir. Bu çalışma fareembriyonik korteks ve serebellumundan alınan nörosferinetanole maruz kaldığında morfolojideki farklılığı araştır-mak için planlandı. Etanol dozu aşırı alkol alımındaki kan konsantrasyonuna benzer idi. Gereç ve yöntem: Gebe albino fare (E14) embriyosunun lateral ventrikül ve dördüncü ventrikülü çevresindeki germinal doku alındı. Primer kültürler, ortam hormon karışımındaki yeterli yoğunluktaki hücreler kullanılarak ekim yapıldı. Kırksekiz saat sonra kültür ortamına etanol (düşük doz = 80 mg/dl; yüksek doz = 400 mg/dl) eklendi. Farklı grupların nörosferleri faz-kontrast mikroskop, triptan mavısı dışlama testi ve 4’-6-Diamidino-2-fenilindol immunoboyama ile analiz edildi. Bulgular: Düşük doz etanol ile kortikal nörosferde %6,9 azalma gözlenirken, serebellar nörosferde %51,2 azalma oldu. Ortalama piknotik hücre sayıları hem düşük hem de yüksek doz etanol uygulanan gruplarda, hem kortikal hem de serebellar kültürlerde, kontrol gruplarına göre anlamlı artış gösterdi (p

E14 fare embriyosundan izole edilen serebellar ve kortikal nörosferler üzerine etanol’ün nörotoksik etkisinin değerlendirilmesi

Objectives: Ethanol exposure during pregnancy has tox-ic effects 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 es-tablished. This study was planned to find out the differ-ence in 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 around the lateral ventricle and fourth ventricle of the embryo of pregnant albino mice (E14) were dissected out. Primary cultures were plated using adequate density of cells in media hormone mix (MHM) and growth factors. After 48 hours, ethanol was added to the cultures (low dose = 80 mg/dl; high dose = 400 mg/dl). NS from different groups were analyzed using phase-contrast microscopy, Trypan blue exclusion assay and immunostaining with 4’-6-Di- amidino-2-phenylindol (DAPI). Results: With low dose ethanol treatment, the mean area of cortical neurospheres was reduced by 6.9% where- as that of the cerebellar neurospheres was reduced by 51.2%. The mean count of pyknotic cells increased signif- icantly (p

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