Rahman H, Lee K, Kentroti S, Vernadakis A
Department of Psychiatry, University of Colorado School of Medicine, Denver 80262.
Neurochem Res. 1993 May;18(5):551-7. doi: 10.1007/BF00966930.
Neuronal-enriched cultures were prepared from 8-day-old chick embryo cerebral hemispheres and exposed to ethanol (50 mM) from day 4 to 8 in culture. At day 8, both control and ethanol-treated cultures were processed for [3H]choline uptake in situ. Uptake was performed on cultures containing either Na(+)-plus or Na(+)-free (Li+) HEPES buffer. Total choline uptake as well as Na(+)-dependent and Na(+)-independent choline uptake were calculated. The Km and Vmax were calculated using the Lineweaver-Burke analysis. Our analysis of the data revealed that ethanol-treated cultures exhibited two values for Vmax, one similar to that found in control cultures and one significantly lower than controls. No differences were observed in Km values between control and ethanol-treated cultures. We interpret the low Vmax to represent a population of cholinergic neurons which have been arrested at an immature stage as a result of ethanol insult.
从8日龄鸡胚大脑半球制备富含神经元的培养物,并在培养的第4天至第8天暴露于乙醇(50 mM)中。在第8天,对对照和乙醇处理的培养物进行原位[3H]胆碱摄取实验。摄取实验在含有Na(+)加或无Na(+)(Li+)的HEPES缓冲液的培养物上进行。计算总胆碱摄取以及Na(+)依赖性和非Na(+)依赖性胆碱摄取。使用Lineweaver-Burke分析计算Km和Vmax。我们对数据的分析表明,乙醇处理的培养物表现出两个Vmax值,一个与对照培养物中发现的值相似,另一个明显低于对照。对照和乙醇处理的培养物之间的Km值没有差异。我们将低Vmax解释为代表由于乙醇损伤而停滞在未成熟阶段的胆碱能神经元群体。