Maternal ethanol consumption before paternal fertilization: Stimulation of hypocretin neurogenesis and ethanol intake in zebrafish offspring
Maternal ethanol consumption before paternal fertilization: Stimulation of hypocretin neurogenesis and ethanol intake in zebrafish offspring
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DOI：
10.1016/j.pnpbp.2019.109728
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发表时间：
2020-01-10
影响因子：
5.6
通讯作者：
Leibowitz, Sarah F.
中科院分区：
文献类型：
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作者：
Collier, Adam D.;Min, Soe S.;Leibowitz, Sarah F.
大量临床和临床前研究表明，胚胎暴露于乙醇显著影响神经元发育，刺激饮酒和相关行为。在啮齿动物和斑马鱼中，我们的研究表明，胚胎暴露于低剂量乙醇中，除了增加青春期的自愿乙醇摄入量外，还增加了下丘脑下丘脑分泌素（hcrt）神经元的密度，hcrt是一种已知的调节奖励相关行为的神经肽。这里在斑马鱼身上提出的问题是，母体在受孕前摄入乙醇是否也会影响神经元和行为发育，这是临床报告提出的现象，但很少研究。为了确定孕前母体乙醇消耗是否也会影响后代的这些hcrt神经元和行为，我们首先标准化了一种测量AB系成年和幼斑马鱼自愿乙醇消耗的方法，分别给予含有10%或0.1%乙醇的明胶餐。我们发现，咀嚼明胶的次数可以准确地衡量成人的摄入量，也可以强有力地预测血液乙醇水平，同时也是斑马鱼幼虫摄入量的可靠指标。然后，我们使用这种喂养模式和实时成像来检查孕前母亲摄入10%乙醇-明胶与普通明胶相比14天对后代神经元发育的影响。成年雌性转基因HuC: EGFP斑马鱼在受精后28小时（hpf）摄入乙醇对分化的HuC（+）神经元数量没有影响，而成年雌性转基因HuC: EGFP斑马鱼孕前摄入乙醇显著增加了后代的hcrt神经元数量，这一效应在受精后28小时（hpf）观察到，并在受精后6和12天（dpf）得到证实。hcrt神经元的增加主要出现在大脑的左侧，表明乙醇的作用不对称，并且伴随着后代的行为变化，包括在6 dpf时显著增加的新颖诱导的运动活动，但不包括在12 dpf时自愿摄入0.1%乙醇-明胶的运动。值得注意的是，这些乙醇摄入量和孕前乙醇刺激的运动活动的测量与hcrt神经元的数量呈强烈的正相关。这些发现表明，孕前母体酒精摄入会影响后代的大脑和行为，产生与胚胎酒精暴露引起的影响相似的影响，并进一步证明乙醇诱导的hcrt神经发生增加会导致乙醇引起的行为障碍。
There are numerous clinical and pre-clinical studies showing that exposure of the embryo to ethanol markedly affects neuronal development and stimulates alcohol drinking and related behaviors. In rodents and zebrafish, our studies show that embryonic exposure to low-dose ethanol, in addition to increasing voluntary ethanol intake during adolescence, increases the density of hypothalamic hypocretin (hcrt) neurons, a neuropeptide known to regulate reward-related behaviors. The question addressed here in zebrafish is whether maternal ethanol intake before conception also affects neuronal and behavioral development, phenomena suggested by clinical reports but seldom investigated. To determine if preconception maternal ethanol consumption also affects these hcrt neurons and behavior in the offspring, we first standardized a method of measuring voluntary ethanol consumption in AB strain adult and larval zebrafish given gelatin meals containing 10% or 0.1% ethanol, respectively. We found the number of bites of gelatin to be an accurate measure of intake in adults and a strong predictor of blood ethanol levels, and also to be a reliable indicator of intake in larval zebrafish. We then used this feeding paradigm and live imaging to examine the effects of preconception maternal intake of 10% ethanol-gelatin compared to plain-gelatin for 14 days on neuronal development in the offspring. Whereas ethanol consumption by adult female HuC:GFP transgenic zebrafish had no impact on the number of differentiated HuC(+) neurons at 28 h post-fertilization (hpf), preconception ethanol consumption by adult female hcrt:EGFP zebrafish significantly increased the number of hcrt neurons in the offspring, an effect observed at 28 hpf and confirmed at 6 and 12 days post-fertilization (dpf). This increase in hcrt neurons was primarily present on the left side of the brain, indicating asymmetry in ethanol's actions, and it was accompanied by behavioral changes in the offspring, including a significant increase in novelty-induced locomotor activity but not thigmotaxis measured at 6 dpf and also in voluntary consumption of 0.1% ethanol-gelatin at 12 dpf. Notably, these measures of ethanol intake and locomotor activity stimulated by preconception ethanol were strongly, positively correlated with the number of hcrt neurons. These findings demonstrate that preconception maternal ethanol consumption affects the brain and behavior of the offspring, producing effects similar to those caused by embryonic ethanol exposure, and they provide further evidence that the ethanol-induced increase in hcrt neurogenesis contributes to the behavioral disturbances caused by ethanol.