Department of Physiology & Pharmacology, Wake Forest School of Medicine, Winston-Salem, North Carolina.
Department of Behavioral Neuroscience, Oregon National Primate Research Center, Oregon Health Sciences University, Portland, Oregon.
Alcohol Clin Exp Res. 2018 Sep;42(9):1661-1673. doi: 10.1111/acer.13821. Epub 2018 Jul 15.
Recent work with long-term ethanol (EtOH) self-administration in nonhuman primate models has revealed a complex array of behavioral and physiological effects that closely mimic human alcohol abuse. Detailed neurophysiological analysis in these models suggests a myriad of pre- and postsynaptic neurobiological effects that may contribute to the behavioral manifestations of long-term EtOH drinking. The molecular mechanisms regulating presynaptic effects of this chronic EtOH exposure are largely unknown. To this end, we analyzed the effects of long-term EtOH self-administration on the levels of presynaptic SNARE complex proteins in Macaca mulatta basolateral amygdala, a brain region known to regulate both aversive and reward-seeking behaviors.
Basolateral amygdala samples from control and EtOH-drinking male and female monkeys were processed. Total basolateral amygdala protein was analyzed by Western blotting using antibodies directed against both core SNARE and SNARE-associated proteins. We also performed correlational analyses between protein expression levels and a number of EtOH drinking parameters, including lifetime grams of EtOH consumed, preference, and blood alcohol concentration.
Significant interactions or main effects of sex/drinking were seen for a number of SNARE core and SNARE-associated proteins. Across the range of EtOH-drinking phenotypes, SNAP25 and Munc13-1 proteins levels were significantly different between males and females, and Munc13-2 levels were significantly lower in animals with a history of EtOH drinking. A separate analysis of very heavy-drinking individuals revealed significant decreases in Rab3c (females) and complexin 2 (males).
Protein expression analysis of basolateral amygdala total protein from controls and animals following long-term EtOH self-administration suggests a number of alterations in core SNARE or SNARE-associated components that could dramatically alter presynaptic function. A number of proteins or multiprotein components were also correlated with EtOH drinking behavior, which suggest a potentially heritable role for presynaptic SNARE proteins.
最近在非人类灵长类动物模型中进行的长期乙醇（EtOH）自我给药研究揭示了一系列与人类酒精滥用密切相关的复杂行为和生理效应。这些模型中的详细神经生理学分析表明，存在许多突触前和突触后神经生物学效应，这些效应可能导致长期 EtOH 饮酒的行为表现。调节这种慢性 EtOH 暴露突触前效应的分子机制在很大程度上是未知的。为此，我们分析了长期 EtOH 自我给药对猕猴基底外侧杏仁核突触前 SNARE 复合物蛋白水平的影响，该脑区已知调节厌恶和寻求奖励行为。
处理来自对照和 EtOH 饮酒的雄性和雌性猴子的基底外侧杏仁核样本。使用针对核心 SNARE 和 SNARE 相关蛋白的抗体通过 Western blot 分析总基底外侧杏仁核蛋白。我们还进行了蛋白表达水平与许多 EtOH 饮酒参数之间的相关性分析，包括终生摄入的 EtOH 克数、偏好和血液酒精浓度。
在许多 SNARE 核心和 SNARE 相关蛋白中，观察到性别/饮酒的显著相互作用或主要效应。在一系列 EtOH 饮酒表型中，SNAP25 和 Munc13-1 蛋白水平在雄性和雌性之间存在显著差异，而具有 EtOH 饮酒史的动物中的 Munc13-2 水平明显较低。对非常大量饮酒者的单独分析显示 Rab3c（女性）和复合蛋白 2（男性）显著减少。
从长期 EtOH 自我给药后的对照动物和动物的基底外侧杏仁核总蛋白的蛋白表达分析表明，核心 SNARE 或 SNARE 相关成分的许多改变可能会极大地改变突触前功能。一些蛋白或多蛋白成分也与 EtOH 饮酒行为相关，这表明突触前 SNARE 蛋白可能具有潜在的遗传性作用。
