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

Abstract

Objectives: Ethanol exposure during pregnancy has toxic effects on the neural stem cells of the developing brain. Differential effects of the alcohol on the neural precursor cells from different regions of the brain have not been established. This study was planned to find out the difference in the morphology of the neurosphere (NS) from the embryonic mouse cortex and cerebellum when exposed to ethanol. The dose of ethanol was comparable to blood alcohol 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-Diamidino-2-phenylindol (DAPI). Results: With low dose ethanol treatment, the mean area of cortical neurospheres was reduced by 6.9% whereas that of the cerebellar neurospheres was reduced by 51.2%. The mean count of pyknotic cells increased significantly (p

Keywords

• Neural stem cell, alcohol ethyl, in vitro toxicity

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

Öz

Amaç: Gebelikte etanol maruziyeti, gelişen beyindeki nöral kök hücreler üzerine toksik etkilere sahiptir. Beynin değişik bölgelerindeki nöral öncül hücreler üzerine alkolün farklı etkileri henüz belirlenmemiştir. Bu çalışma fare embriyonik korteks ve serebellumundan alınan nörosferin etanole maruz kaldığında morfolojideki farklılığı araştırmak 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

Anahtar Kelimeler

• Nöral kök hücre, etil alkol, in vitro toksisite

Kaynakça

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