Jenab S, Inturrisi C E
Department of Pharmacology, Cornell University Medical College, New York, NY 10021.
Brain Res Mol Brain Res. 1994 Nov;27(1):95-102. doi: 10.1016/0169-328x(94)90189-9.
We have used a sensitive solution hybridization assay with a riboprobe transcribed from the coding sequence of the delta opioid receptor (DOR) to quantitate the changes in DOR mRNA transcript levels following exposure of NG108-15 cells to ethanol and/or the opioid antagonist, naloxone. Incubation of NG108-15 cells with 200 mM ethanol or 1 microM naloxone, treatments that have previously been shown to upregulate DOR binding, increased DOR mRNA transcript levels 2 to 3 fold. DOR mRNA levels peaked at 24 to 48 h after exposure to either ethanol or naloxone. At 168 h, DOR mRNA levels in NG108-15 cells exposed to naloxone had returned to control (untreated) levels while the levels in ethanol treated cells remained nearly equal to peak values. Exposure to a combination of ethanol plus naloxone for 24 h produced an additive effect, so that DOR mRNA transcripts were increased 3 fold. Northern blot analysis identified six DOR transcript bands ranging in size from 8.7 to 2.1 kb. The above treatments increased each of the six bands proportionately, so that no difference was observed in the fraction of the total hybridization signal produced by each band of the Northern blot. These results demonstrate that each of the DOR transcripts in NG108-15 cells are subject to homologous (naloxone) as well as heterologous (ethanol) upregulation.
我们使用了一种灵敏的溶液杂交检测方法，该方法采用从δ阿片受体（DOR）编码序列转录而来的核糖探针，来定量NG108 - 15细胞暴露于乙醇和/或阿片拮抗剂纳洛酮后DOR mRNA转录水平的变化。用200 mM乙醇或1 μM纳洛酮孵育NG108 - 15细胞，先前已证明这些处理可上调DOR结合，使DOR mRNA转录水平增加2至3倍。暴露于乙醇或纳洛酮后，DOR mRNA水平在24至48小时达到峰值。在168小时时，暴露于纳洛酮的NG108 - 15细胞中的DOR mRNA水平已恢复到对照（未处理）水平，而乙醇处理细胞中的水平仍几乎等于峰值。暴露于乙醇加纳洛酮的组合24小时产生了累加效应，使得DOR mRNA转录本增加了3倍。Northern印迹分析鉴定出六个DOR转录带，大小范围从8.7至2.1 kb。上述处理使六个带中的每一个都成比例增加，因此在Northern印迹的每个带产生的总杂交信号分数中未观察到差异。这些结果表明，NG108 - 15细胞中的每个DOR转录本都受到同源（纳洛酮）以及异源（乙醇）上调的影响。
