Details

Autor(en) / Beteiligte

• Cherian, Priscilla P.
• Schenker, Steven
• Henderson, George I.

Titel

Ethanol-Mediated DNA Damage and PARP-1 Apoptotic Responses in Cultured Fetal Cortical Neurons

Ist Teil von

• Alcoholism, clinical and experimental research, 2008-11, Vol.32 (11), p.1884-1892

Ort / Verlag

Oxford, UK: Blackwell Publishing Ltd

Erscheinungsjahr

2008

Link zum Volltext

Quelle

Wiley Online Library Journals【キャンパス外アクセス可】

Beschreibungen/Notizen

• Background:  Prior studies by many laboratories have illustrated that ethanol can elicit a cascade of caspase‐dependent apoptotic events in cultured neurons. Studies in our laboratory have connected this to oxidative stress and effects on fetal cortical neuron glutathione homeostasis. Aims:  The intent of the following studies is to address mechanisms underlying ethanol‐associated DNA damage that may be connected to apoptotic death of neurons. Methods:  Cultures of fetal rat cerebral cortical neurons were utilized. Estimates of DNA damage was determined by Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining and nuclear condensation; Poly(ADP‐ribose) polymerase‐1 (PARP‐1) expression was determined by immunostaining and Western blotting; and occurrence of parylation and AIF translocations were assessed by Western blotting. Results:  Ethanol treatment of the neurons generated increases in DNA damage by 4 hours while nuclear condensation was low at the short exposure period but increased markedly by 24 hours. This was temporally related to a marked up‐regulation of PARP‐1 expression. Activity of PARP‐1, as assessed by PolyADP‐ribose (PAR) formation, occurred within 15 minutes and peaked by 6 to 8 hours of ethanol treatment. An almost complete translocation of apoptosis inducing factor (AIF) from mitochondria to the nucleus occurred by 24 hours of ethanol treatment (4.0 mg/ml). Ethanol treatment for 4, 12, and 24 hours elicited an increasing caspase‐mediated cleavage of PARP‐1 to its 24 kDa fragment. Conclusions:  These data illustrate the rapid occurrence of DNA damage following ethanol exposure and that PARP‐1 pathways may play a role in the subsequent apoptotic death of these neurons.