Qian Kun, Jiang Chengyun, Guan Daojie, Zhuang Anxiang, Meng Xiangkun, Wang Jianjun
College of Plant Protection, Yangzhou University, Yangzhou 225009, China.
College of Plant Protection, Jilin Agricultural University, Changchun 130118, China.
Insects. 2023 Jun 25;14(7):580. doi: 10.3390/insects14070580.
The glutamate-gated chloride channels (GluCls) play essential roles in signal transduction by regulating fast inhibitory synaptic transmission in the nervous system of invertebrates. While there is only one GluCl subunit in the insect, the diversity of insect GluCls is broadened by alternative splicing. In the present study, three variant genes were cloned from the red flour beetle . Analysis of the characteristics of TcGluCls including sequence features, genomic structures, and alternative splicing revealed that TcGluCls had the typical structural features of GluCls and showed high homologies with the GluCls from other insect orders. The TcGluCl-encoding gene consists of nine exons and three variants (, , and ) were generated by the alternative splicing of exon 3, which was a highly conserved alternative splicing site in insect GluCls. Homology modeling of TcGluCl-3a showed that the exon 3 coding protein located at the N-terminal extracellular domain, and there were no steric clashes encountered between the exon 3 coding region and ivermectin/glutamate binding pocket, which indicated that the alternative splicing of exon 3 might have no impact on the binding of GluCls to glutamate and insecticide. In addition to the head tissue, and also had high expressions in the ovary and testis of , whereas showed high expression in the midgut, suggesting the diverse physiological functions of TcGluCl variants in . The total and three variants showed the highest expression levels in the early stage larvae. The expressions of , , and were significantly increased from the late-stage larvae to the early stage pupae and indicated that the might be involved in the growth and development of . These results are helpful to further understand the molecular characteristics of insect GluCls and provide foundations for studying the specific function of the GluCl variant.
谷氨酸门控氯离子通道（GluCls）通过调节无脊椎动物神经系统中的快速抑制性突触传递，在信号转导中发挥着重要作用。虽然昆虫中只有一个GluCl亚基，但可变剪接拓宽了昆虫GluCls的多样性。在本研究中，从赤拟谷盗中克隆了三个变异基因。对赤拟谷盗GluCls（TcGluCls）的特性进行分析，包括序列特征、基因组结构和可变剪接，结果表明，TcGluCls具有GluCls的典型结构特征，并且与其他昆虫目的GluCls具有高度同源性。编码TcGluCl的基因由九个外显子组成，外显子3的可变剪接产生了三个变异体（、和），外显子3是昆虫GluCls中一个高度保守的可变剪接位点。TcGluCl-3a的同源建模表明，外显子3编码的蛋白质位于N端细胞外结构域，外显子3编码区与伊维菌素/谷氨酸结合口袋之间没有空间冲突，这表明外显子3的可变剪接可能对GluCls与谷氨酸和杀虫剂的结合没有影响。除头部组织外，和在赤拟谷盗的卵巢和睾丸中也有高表达，而在中肠中高表达，这表明TcGluCl变异体在赤拟谷盗中具有多种生理功能。总和三个变异体在幼虫早期表达水平最高。、和的表达从幼虫后期到蛹早期显著增加，表明可能参与了赤拟谷盗的生长和发育。这些结果有助于进一步了解昆虫GluCls的分子特征，并为研究GluCl变异体的具体功能提供基础。
