Differential involvement of glutamate-gated chloride channel splice variants in the olfactory memory processes of the honeybee Apis mellifera
Differential involvement of glutamate-gated chloride channel splice variants in the olfactory memory processes of the honeybee Apis mellifera
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DOI：
10.1016/j.pbb.2014.05.025
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发表时间：
2014-09-01
影响因子：
3.6
通讯作者：
Armengaud, Catherine
中科院分区：
文献类型：
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作者：
Demares, Fabien;Drouard, Florian;Armengaud, Catherine
谷氨酸门控氯离子通道（GluCl）属于cys环配体门控离子通道超家族，在多种无脊椎动物神经系统中均有表达。在蜜蜂中，独特的基因amel_glucl编码两个可变剪接亚基Amel_GluCl A和Amel_GluCl B。这些变异体在蜜蜂脑中的表达和差异定位以前已有报道。在这里，我们的特点是参与嗅觉学习和记忆过程中的每个变体，使用特定的小干扰RNA（siRNA）针对每个变体。首先，在mRNA和蛋白质水平上测试了两种siRNA降低其靶标表达的功效。两种蛋白质在注射后24 h显示其各自的表达降低。其次，将每种siRNA注射到大脑中，以通过使用调节长鼻延伸反射（PER）的经典范例来测试其是否影响嗅觉记忆。发现Amel_GluCl A仅参与1-壬醇的回收，而Amel_GluCl B参与对用作条件刺激或新气味剂的2-己醇的PER反应。在此，首次在无脊椎动物物种中表征了两种高度相似的GluCl亚基的差异行为参与。(C)2014爱思唯尔公司All rights reserved.
Glutamate-gated chloride channels (GluCl) belong to the cys-loop ligand-gated ion channel superfamily and their expression had been described in several invertebrate nervous systems. In the honeybee, a unique gene amel_glucl encodes two alternatively spliced subunits, Amel_GluCl A and Amel_GluCl B. The expression and differential localization of those variants in the honeybee brain had been previously reported. Here we characterized the involvement of each variant in olfactory learning and memory processes, using specific small-interfering RNA (siRNA) targeting each variant. Firstly, the efficacy of the two siRNAs to decrease their targets' expression was tested, both at mRNA and protein levels. The two proteins showed a decrease of their respective expression 24 h after injection. Secondly, each siRNA was injected into the brain to test whether or not it affected olfactory memory by using a classical paradigm of conditioning the proboscis extension reflex (PER). Amel_GluCl A was found to be involved only in retrieval of 1-nonanol, whereas Amel_GluCl B was involved in the PER response to 2-hexanol used as a conditioned stimulus or as new odorant Here for the first time, a differential behavioral involvement of two highly similar GluCl subunits has been characterized in an invertebrate species. (C) 2014 Elsevier Inc. All rights reserved.