Identification of Slit3 as a locus affecting nicotine preference in zebrafish and human smoking behaviour
Identification of Slit3 as a locus affecting nicotine preference in zebrafish and human smoking behaviour
复制标题
鉴定 Slit3 作为影响斑马鱼尼古丁偏好和人类吸烟行为的基因座
DOI：
10.1101/453928
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
García-González J
中科院分区：
文献类型：
--
作者：
García-González J
背景虽然有明确的证据表明尼古丁成瘾的易感性与遗传有关，但从人类研究中识别因果等位基因和途径已被证明是困难的。模式物种的突变产生野生型种群中不存在的强烈表型，并可用于确定可量化行为的生物学机制。我们验证了斑马鱼尼古丁偏好的正向遗传筛选可以预测影响人类吸烟行为的基因座和生物学机制的假设。方法基于群体的乙基亚硝脲诱变斑马鱼正向遗传筛选被用来识别表现出尼古丁偏好改变的鱼类线条。采用免疫组织化学、行为学和定量聚合酶链式反应等方法对突变幼虫进行鉴定。对来自英国和芬兰的一项双胞胎研究中的同源人类基因座进行了集中的SNP分析，评估了斑马鱼数据对人类吸烟行为的预测有效性。结果我们发现，尼古丁偏好在鱼类和人类中都是可遗传的，并发现斑马鱼Slit3基因的功能丧失突变与尼古丁偏好增加有关。Slit3突变幼鱼显示出对多巴胺能拮抗剂的声音惊吓的习惯性改变了敏感性，并增加了Drd2和Drd3mRNA的表达。多巴胺能神经元的寻路未受影响。对两个独立的人类队列中SLIT3基因座的分析确定了预测吸烟水平和戒烟可能性的两个遗传标记。结论这些发现表明SLIT3信号在影响与尼古丁依赖相关的行为的多巴胺能通路的发展中发挥作用，并证实了斑马鱼模型在探索复杂的人类行为方面的翻译相关性。
BACKGROUNDAlthough there is clear evidence of genetic contributions to susceptibility to nicotine addiction, it has proved difficult to identify causal alleles and pathways from studies in humans. Mutagenesis in model species generates strong phenotypes not present in wildtype populations and can be used to identify biological mechanisms underlying quantifiable behaviours. We tested the hypothesis that a forward genetic screen for nicotine preference in zebrafish can predict loci and biological mechanisms influencing human smoking behaviour.METHODSA population-based forward genetic screen of ethylnitrosurea-mutagenized zebrafish was used to identify lines of fish showing altered nicotine preference. Immunohistochemical, behavioral and quantitative PCR analyses were used to characterize mutant larvae. Focussed SNP analysis of the homologous human locus in cohorts from the UK and a Finnish Twin study assessed the predictive validity of the zebrafish data for human smoking behavior.RESULTSWe show nicotine preference is heritable in fish as in humans and identify loss-of-function mutations in the zebrafishSlit3gene as associated with increased nicotine preference.Slit3mutant larval fish showed altered sensitivity of habituation to acoustic startle to dopaminergic antagonists and increasedDrd2andDrd3mRNA expression. Dopaminergic neuronal pathfinding was unaffected. Analysis of the SLIT3 locus in two independent human cohorts identified 2 genetic markers that predict level of cigarette consumption and likelihood of cessation.CONCLUSIONThese findings suggest a role for SLIT3 signaling in development of dopaminergic pathways affecting behaviours associated with nicotine dependence and confirm the translational relevance of the zebrafish model in exploring complex human behaviors.