Biology Department, University of North Dakota, United States; Turtle Mountain Community College, United States.
Pharmacol Biochem Behav. 2012 Dec;103(2):157-67. doi: 10.1016/j.pbb.2012.07.017. Epub 2012 Aug 15.
Finding genetic polymorphisms and mutations linked to addictive behavior can provide important targets for pharmaceutical and therapeutic interventions. Forward genetic approaches in model organisms such as zebrafish provide a potentially powerful avenue for finding new target genes. In order to validate this use of zebrafish, the molecular nature of its reward system must be characterized. We have previously reported the use of cocaine-induced conditioned place preference (CPP) as a reliable method for screening mutagenized fish for defects in the reward pathway. Here we test if CPP in zebrafish involves the dopaminergic system by co-treating fish with cocaine and dopaminergic antagonists. Sulpiride, a potent D2 receptor (DR2) antagonist, blocked cocaine-induced CPP, while the D1 receptor (DR1) antagonist SCH23390 had no effect. Acute cocaine exposure also induced a rise in the expression of tyrosine hydroxylase (TH), an important enzyme in dopamine synthesis, and a significant decrease in the expression of elongation factor 1α (EF1α), a housekeeping gene that regulates protein synthesis. Cocaine selectively increased the ratio of TH/EF1α in the telencephalon, but not in other brain regions. The cocaine-induced change in TH/EF1α was blocked by co-treatment with sulpiride, but not SCH23390, correlating closely with the action of these drugs on the CPP behavioral response. Immunohistochemical analysis revealed that the drop in EF1α was selective for the dorsal nucleus of the ventral telencephalic area (Vd), a region believed to be the teleost equivalent of the striatum. Examination of TH mRNA and EF1α transcripts suggests that regulation of expression is post-transcriptional, but this requires further examination. These results highlight important similarities and differences between zebrafish and more traditional mammalian model organisms.
发现与成瘾行为相关的遗传多态性和突变,可以为药物和治疗干预提供重要的靶点。模式生物(如斑马鱼)中的正向遗传学方法为寻找新的靶基因提供了一个潜在的有力途径。为了验证这种斑马鱼的应用,必须对其奖励系统的分子性质进行特征分析。我们之前曾报道过,使用可卡因诱导的条件位置偏好(CPP)作为筛选突变鱼在奖励途径缺陷的可靠方法。在这里,我们通过与可卡因和多巴胺拮抗剂共同处理鱼来测试 CPP 在斑马鱼中是否涉及多巴胺能系统。Sulpiride 是一种有效的 D2 受体(DR2)拮抗剂,可阻断可卡因诱导的 CPP,而 D1 受体(DR1)拮抗剂 SCH23390 则没有作用。急性可卡因暴露还诱导了酪氨酸羟化酶(TH)的表达增加,TH 是多巴胺合成中的重要酶,同时也导致延伸因子 1α(EF1α)的表达显著下降,EF1α 是一种调节蛋白质合成的管家基因。可卡因选择性地增加了脑端脑的 TH/EF1α 比值,但在其他脑区则没有。Sulpiride 共同处理可阻断可卡因诱导的 TH/EF1α 变化,但 SCH23390 则没有,这与这些药物对 CPP 行为反应的作用密切相关。免疫组织化学分析显示,EF1α 的下降是腹侧端脑背核(Vd)的选择性下降,该区域被认为是硬骨鱼的纹状体等效物。对 TH mRNA 和 EF1α 转录本的检查表明,表达的调节是转录后,但这需要进一步检查。这些结果突出了斑马鱼与更传统的哺乳动物模型生物之间的重要相似性和差异。