The Hospital for Sick Children, Toronto, Ontario, Canada.
Department of Psychology, University of Toronto, Mississauga, Ontario, Canada.
Prog Neuropsychopharmacol Biol Psychiatry. 2021 Aug 30;110:110327. doi: 10.1016/j.pnpbp.2021.110327. Epub 2021 Apr 14.
The zebrafish is increasingly well utilized in alcohol research, particularly in modeling human fetal alcohol spectrum disorders (FASD). FASD results from alcohol reaching the developing fetus intra utero, a completely preventable yet prevalent and devastating life-long disorder. The hope with animal models, including the zebrafish, is to discover the mechanisms underlying this disease, which may aid treatment and diagnosis. In the past, we developed an embryonic alcohol exposure regimen that is aimed at mimicking the milder, and most prevalent, forms of FASD in zebrafish. We have found numerous lasting alterations in behavior, neurochemistry, neuronal markers and glial cell phenotypes in this zebrafish FASD model. Using the same model (2 h long bath immersion of 24 h post-fertilization old zebrafish eggs into 1% vol/vol ethanol), here we conduct a proof of concept analysis of voltage-gated cation channels, investigating potential embryonic alcohol induced changes in L-, T- and N- type Ca and the SCN1A Na channels using Western blot followed by immunohistochemical analysis of the same channels in the pallium and cerebellum of the zebrafish brain. We report significant reduction of expression in all four channel proteins using both methods. We conclude that reduced voltage-gated cation channel expression induced by short and low dose exposure to alcohol during embryonic development of zebrafish may contribute to the previously demonstrated lasting behavioral and neurobiological changes.
斑马鱼越来越多地被用于酒精研究，特别是在模拟人类胎儿酒精谱系障碍（FASD）方面。FASD 是由于酒精进入胎儿体内，这是一种完全可以预防但普遍存在且具有破坏性的终身疾病。包括斑马鱼在内的动物模型的希望是发现这种疾病的机制，这可能有助于治疗和诊断。过去，我们开发了一种胚胎期酒精暴露方案，旨在模拟斑马鱼中较轻且最常见的 FASD 形式。我们在这种斑马鱼 FASD 模型中发现了许多持久的行为、神经化学、神经元标志物和神经胶质细胞表型的改变。使用相同的模型（24 小时后受精的斑马鱼卵在 1%体积/体积乙醇中浸泡 2 小时长的浴），我们在这里进行了电压门控阳离子通道的概念验证分析，使用 Western blot 研究潜在的胚胎酒精诱导的 L、T 和 N 型 Ca 和 SCN1A Na 通道变化，随后对斑马鱼大脑的大脑皮层和小脑进行相同通道的免疫组织化学分析。我们使用这两种方法都报告了所有四种通道蛋白表达的显著减少。我们得出结论，短时间和低剂量暴露于酒精会导致斑马鱼胚胎发育过程中电压门控阳离子通道表达减少，这可能导致之前证明的持久行为和神经生物学变化。
