The Department of Biobehavioral Health, Pennsylvania State University, University Park, PA, USA.
Department of Neuroscience, Perelman School of Medicine, University of Pennsylvania Medical School, Philadelphia, PA, USA.
Neuropharmacology. 2018 Oct;141:192-200. doi: 10.1016/j.neuropharm.2018.08.019. Epub 2018 Aug 28.
Numerous studies have attributed the psychopathology of anxiety and stress disorders to maladaptive behavioral responses such as an inability to extinguish fear. Therefore, understanding neural substrates of fear extinction is imperative for developing more effective therapies for anxiety and stress disorders. Although several studies indicated a role for cholinergic transmission and nicotinic acetylcholine receptors (nAChRs) in anxiety and stress disorder symptomatology, very little is known about the specific contribution of nAChRs in the fear extinction process. In the present study, we first examined the involvement of several brain regions essential for fear extinction (i.e., dorsal and ventral hippocampus, dHPC and vHPC; infralimbic, IL, and prelimbic, PL of the medial prefrontal cortex, mPFC; basolateral nucleus of the amygdala, BLA) in the impairing effects of a nAChR agonist, nicotine, on contextual fear extinction in mice. Our results showed that systemic administration of nicotine during contextual fear extinction increased c-fos expression in the vHPC and BLA while not affecting dHPC, IL or PL. In line with these results, local nicotine infusions into the vHPC, but not dHPC, resulted in impaired contextual fear extinction. Interestingly, we found that local nicotine infusions into the PL also resulted in impairment of contextual fear extinction. Second, we measured the protein levels of the GABA synthesizing enzymes GAD65 and GAD67 in the dHPC and vHPC during contextual fear extinction. Our results showed that in the group that received acute nicotine, both GAD65 and GAD67 protein levels were downregulated in the vHPC, but not in dHPC. This effect was negatively correlated with the level of freezing response during fear extinction suggesting that the downregulated GAD65/67 levels were associated with disrupted fear extinction. Finally, using c-fos/GAD65/67 double immunofluorescence, we showed that nicotine mainly increased c-fos expression in non-GABAergic ventral hippocampal cells, indicating that acute nicotine increases vHPC excitability. Overall, our results suggest that acute nicotine's impairing effects on fear extinction are associated with ventral hippocampal disinhibition. Therefore, these results further our understanding of the interaction between nicotine addiction and anxiety and stress disorders by describing novel neural mechanisms mediating fear extinction.
许多研究将焦虑和应激障碍的精神病理学归因于适应不良的行为反应,例如无法消除恐惧。因此,了解恐惧消退的神经基础对于开发更有效的焦虑和应激障碍治疗方法至关重要。尽管有几项研究表明胆碱能传递和烟碱型乙酰胆碱受体(nAChRs)在焦虑和应激障碍症状学中起作用,但对于 nAChRs 在恐惧消退过程中的具体作用知之甚少。在本研究中,我们首先研究了几个对恐惧消退至关重要的脑区(即背侧和腹侧海马体、dHPC 和 vHPC;内侧前额叶皮质的眶额回和前额叶皮层、mPFC;杏仁核的基底外侧核、BLA)在烟碱型乙酰胆碱受体激动剂尼古丁对小鼠情境性恐惧消退的损害作用中的参与情况。我们的结果表明,在情境性恐惧消退期间,系统给予尼古丁会增加 vHPC 和 BLA 中的 c-fos 表达,而不会影响 dHPC、IL 或 PL。与这些结果一致的是,局部给予烟碱到 vHPC 会导致情境性恐惧消退受损,但给予 dHPC 则不会。有趣的是,我们发现局部给予烟碱到 PL 也会导致情境性恐惧消退受损。其次,我们在情境性恐惧消退期间测量了 dHPC 和 vHPC 中的 GABA 合成酶 GAD65 和 GAD67 的蛋白水平。我们的结果表明,在接受急性尼古丁的组中,vHPC 中的 GAD65 和 GAD67 蛋白水平均下调,但 dHPC 中则没有。这种效应与恐惧消退期间的冻结反应水平呈负相关,表明下调的 GAD65/67 水平与恐惧消退受损有关。最后,通过 c-fos/GAD65/67 双重免疫荧光,我们发现尼古丁主要增加非 GABA 能性腹侧海马细胞中的 c-fos 表达,表明急性尼古丁增加 vHPC 的兴奋性。总的来说,我们的结果表明,急性尼古丁对恐惧消退的损害作用与腹侧海马抑制作用的破坏有关。因此,这些结果通过描述介导恐惧消退的新的神经机制,进一步了解了尼古丁成瘾与焦虑和应激障碍之间的相互作用。