Nilesh Kumar MİTRA, Kanakeswary KRİSHNAN, Chong Chung HİONG, Yen Nee DİNG, Hsiao Lung EDDİE, Archana SİKARWAR
E14 fare embriyosundan izole edilen serebellar ve kortikal nörosferler üzerine etanol\'ün nörotoksik etkisinin değerlendirilmesi
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
Evaluation of the neurotoxic effects of ethanol on the cerebellar and cortical neurospheres isolated from E14 mouse embryo
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,
___
- Lemoine P, Harousseau H, Borteyru JP, Menuet JC. Children of alcoholic parents-observed anomalies: discussion of 127 cases. Ther Drug Monit 2003;25 (2):132-6.
- Jones KL, Smith DW. Recognition of the fetal alcohol syndrome in early infancy. Lancet 1973; 302 (7836): 999-1001.
- Chen WJ, Parnell SE, West JR. Neonatal alcohol and nicotine exposure limits brain growth and depletes cerebellar Purkinje cells. Alcohol 1998;15(1):33-41.
- Coulter CL, Leech RW, Schaefer GB, Scheithauer BW, Brumback RA. Midline cerebral dysgenesis, dysfunc- tion of the hypothalamic-pituitary axis, and fetal alco- hol effects. Arch Neurol 1993;50 (7):771-5.
- Chen WJ, Parnell SE, West JR. Effects of alcohol and nicotine on developing olfactory bulb: loss of mitral cells and alterations in neurotransmitter levels. Alco- hol Clin Exp Res 1999;23(1):18-25.
- Streissguth AP and O’Malley K. Neuropsychiatric im- plications and long-term consequences of fetal alco- hol spectrum disorders. Semin Clin Neuropsychiatry 2000; 5 (3): 177-90.
- Cebolla AM, Cheron G, Hourez R, Bearzatto B, Dan B, Servais L. Effects of maternal alcohol consumption during breastfeeding on motor and cerebellar Purkinje cells behavior in mice. Neurosci Lett 2009;455 (1):4-7.
- Finlay BL, Darlington RB. Linked regularities in the de- velopment and evolution of mammalian brains. Sci- ence 1995; 268(5217):1578-84.
- Klein C, Butt SJ, Machold RP, Johnson JE, Fishell G. Cerebellum- and forebrain-derived stem cells possess intrinsic regional character. Development 2005;132(20):4497-508.
- National Institute of Alcohol Abuse Alcoholism. NIAAA Council Approves Definition of Binge Drinking. NIAAA newsletter[serialonline]2004,http://pubs.niaaa.nih. gov/publications/newsletter/winter2004/newsletter_ number3.pdf
- Sulik KK, Lauder JM, Dehart DB. Brain malformations in prenatal mice following acute maternal ethanol ad- ministration. Int J Dev Neurosc 1984;2(3): 203-14.
- Chen WJ, Maier SE, Parnell SE, West JR. Alcohol and the developing brain: neuroanatomical studies. Alcohol Res Health 2003; 27 (2): 174-80.
- Goodlett CR, Horn KH. Mechanisms of alcohol- induced damage to the developing nervous system. Alcohol Res Health 2001;25 (3):175-84.
- Chojnacki A, Weiss S. Production of neurons, astro- cytes and oligodendrocytes from mammalian CNS stem cells. Nat Protoc 2008; 3(6):935-40.
- Lee A, Kessler JD, Read TA et al. Isolation of neural stem cells from the postnatal cerebellum. Nat Neuro- sci 2005;8(6):723-9.
- Anthony B, Zhou FC, Ogawa T, Goodlett CR, Ruiz J. Alcohol exposure alters cell cycle and apoptotoic events during early neurulation. Alcohol Alcohol 2008;43(3):261-73.
- Zharkovsky T, Kaasik A, Jaako K, Zharkovsky A. Neu- rodegeneration and production of the new cells in the dentate gyrus of juvenile rat hippocampus after a sin- gle administration of ethanol. Brain Res 2003;978(1- 2):115-23.
- Aberg E, Hofstetter CP, Olson L, Brené S. Moderate ethanol consumption increases hippocampal cell pro- liferation and neurogenesis in the adult mouse. Int J Neuropsychopharmacol 2005;8(4):557-67.
- Diaz B, Serna F, De Pablo F, de la Rosa EJ. In vivo regulation of cell death by embryonic (pro) insulin and the insulin receptor during early retinal neurogenesis. Development 2000;127(8):1641-9.
- Kuan CY, Schloemer AJ, Lu A et al. Hypoxia-ischemia induces DNA synthesis without cell proliferation in dy- ing neurons in adult rodent brain. J Neurosci 2004; 24(47):10763-72.
- Cherian PP, Schenker S, Henderson GI. Ethanol-me- diated DNA damage and PARP-1 apoptotic responses in cultured fetal cortical neurons. Alcohol Clin Exp Res 2008;32(11):1884-92.
- Başlangıç: 2010
- Yayıncı: Sağlık Araştırmaları Derneği
Sayıdaki Diğer Makaleler
Nilesh Kumar MİTRA, Kanakeswary KRİSHNAN, Chong Chung HİONG, Yen Nee DİNG, Hsiao Lung EDDİE, Archana SİKARWAR
Kahramanmaraş'ta gerçekleşen toplu kardeş intiharlarının incelenmesi
Ramazan Karanfil, Alper KETEN, Cem Zeren, Mustafa Haki Sucaklı
Multipl sklerozlu hastalarda serum prolidaz ve iskemi modifiye albümin düzeyleri
Mehmet Uğur Çevik, Yavuz Yücel, Adalet Arıkanoğlu, Sefer Varol, Eşref Akıl, Hatice Yüksel, Mehmet Ufuk Aluçlu
Julide Sedef GÖÇMEN, Ünase BÜYÜKKOÇAK, Osman CAĞLAYAN