Chien Ya-Chih, Lin Shing-Hong, Lien Cheng-Chang, Wood John N, Chen Chih-Cheng
Program in Molecular Medicine, National Yang Ming Chiao Tung University and Academia Sinica, Taipei, Taiwan.
Institute of Biomedical Sciences, Academia Sinica, Taipei, Taiwan.
J Biomed Sci. 2025 Apr 22;32(1):43. doi: 10.1186/s12929-025-01138-6.
Anxiety is an innate response in the face of danger. When anxiety is overwhelming or persistent, it could be considered an anxiety disorder. Recent studies have shown that acid-sensing ion channels (ASICs) represent a novel class of promising targets for developing effective therapies for anxiety. Especially, ASIC1a and ASIC4 of the ASIC family are widely expressed in the central nervous system and their gene knockouts result in reducing or enhancing anxiety-like responses in mice respectively. However, how ASIC1a and ASIC4 modulate anxiety-associated responses remains unknown.
Here we combined chemo-optogenetic, conditional knockout, gene rescue, molecular biology and biochemistry, and electrophysiological approaches to probe the roles of ASIC4 and ASIC4-expressing cells in anxiety-associated responses in mouse models.
Chemo-optogenetically activating ASIC4-positive cells induced fear and anxiety-like responses in mice. Also, mice lacking ASIC4 (Asic4) in the amygdala or the bed nucleus of the stria terminalis (BNST) exhibited anxiety-associated phenotypes. Conditional knockout of ASIC1a in ASIC4-positive cells reduced anxiety-associated behaviors. In situ hybridization analyses indicated that ASIC4 transcripts were highly co-localized with ASIC1a in the amygdala and BNST. We identified two glycosylation sites of ASIC4, Asn191 and Asn341, that were involved in interacting with ASIC1a and thus could modulate ASIC1a surface protein expression and channel activity. More importantly, viral vector-mediated gene transfer of wild-type ASIC4 but not Asn191 and Asn341 mutants in the amygdala or BNST rescued the anxiogenic phenotypes of Asic4 mice.
Together, these data suggest that ASIC4 plays an important role in fear and anxiety-related behaviors in mice by modulating ASIC1a activity in the amygdala and BNST.
焦虑是面对危险时的一种本能反应。当焦虑情绪压倒性存在或持续时，可被视为焦虑症。最近的研究表明，酸敏感离子通道（ASICs）是开发有效焦虑症治疗方法的一类新的有前景的靶点。特别是，ASIC家族的ASIC1a和ASIC4在中枢神经系统中广泛表达，它们的基因敲除分别导致小鼠焦虑样反应减少或增强。然而，ASIC1a和ASIC4如何调节与焦虑相关的反应仍不清楚。
在这里，我们结合化学光遗传学、条件性敲除、基因拯救、分子生物学和生物化学以及电生理方法，来探究ASIC4和表达ASIC4的细胞在小鼠模型中与焦虑相关反应中的作用。
化学光遗传学激活ASIC4阳性细胞可诱导小鼠产生恐惧和焦虑样反应。此外，杏仁核或终纹床核（BNST）中缺乏ASIC4（Asic4）的小鼠表现出与焦虑相关的表型。在ASIC4阳性细胞中条件性敲除ASIC1a可减少与焦虑相关的行为。原位杂交分析表明，ASIC4转录本在杏仁核和BNST中与ASIC1a高度共定位。我们确定了ASIC4的两个糖基化位点，即天冬酰胺191和天冬酰胺341，它们参与与ASIC1a相互作用，从而可调节ASIC1a表面蛋白表达和通道活性。更重要的是，病毒载体介导的野生型ASIC4而非天冬酰胺191和天冬酰胺341突变体在杏仁核或BNST中的基因转移挽救了Asic4小鼠的致焦虑表型。
总之，这些数据表明ASIC4通过调节杏仁核和BNST中的ASIC1a活性，在小鼠恐惧和焦虑相关行为中起重要作用。
