The mammalian heme oxygenase (HO) plays an important role in cytoprotection against oxidative-stress-induced cell damage; however, functional characterization of insect HO is still limited. In this study, cDNA encoding a HO, named Sf HO, was cloned from Spodoptera frugiperda. Analysis of the transcription level and enzymatic activity showed that exposure of the LC 30 concentration of chlorantraniliprole to the third instar larvae significantly upregulated both the mRNA level and enzymatic activity of Sf HO at 24 h after treatment. Further injection of the HO activator, hemin, into the third instar larvae led to the upregulation of Sf HO as well as decreased susceptibility of S. frugiperda to chlorantraniliprole. Consistently, overexpression of Sf HO increased the Sf9 cell viability under chlorantraniliprole treatment. Strikingly, both RNAi and the dual-luciferase reporter assay in Sf9 cells revealed that, unlike mammalian HO that is regulated by the transcription factor nuclear factor erythroid 2-related factor 2 (Nrf2), Sf HO was not subject to the regulation by cap 'n' collar isoform C (CncC), the Nrf2 homologue in insects. These data provide insights into the function and regulatory mechanism of insect HOs and had applied implications for the control of S. frugiperda.
Chilo suppressalis has developed high levels of resistance to abamectin in many areas of China, while the underline resistance mechanisms are largely unclear. ATP-binding cassette (ABC) transporters function in transporting a large diversity of substrates including insecticides and play important roles in the detoxification metabolism of insects. In this study, synergism bioassay revealed that the ABC transporters were involved in the detoxification of C. suppressalis to abamectin. Six ABC transporter genes were upregulated in C. suppressalis after abamectin exposure, among which five genes CsABCC8, CsABCE1, CsABCF1, CsABCF2, and CsABCH1 were induced in the detoxification-related tissues. In addition, the five ABC transporters were recombinantly expressed in Sf9 cells, and the cytotoxicity assay showed that the viabilities of cells expressing CsABCC8 or CsABCH1 were significantly increased when compared with the viabilities of cells expressing EGFP after abamectin, chlorantraniliprole, cyantraniliprole, fipronil, and chlorpyrifos treatment, respectively. Overexpression of CsABCE1 significantly increased the viabilities of cells to abamectin, chlorantraniliprole, deltamethrin, and indoxacarb exposure, respectively. These results suggested that CsABCC8, CsABCE1, and CsABCH1 might participate in the detoxification and transport of abamectin and several other classes of insecticides in C. suppressalis. Our study provides valuable insights into the transport-related detoxification mechanisms in C. suppressalis and other insects.
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 TcGluCl variant genes were cloned from the red flour beetle Tribolium castaneum. 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 (TcGluCl-3a, TcGluCl-3b, and TcGluCl-3c) 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, TcGluCl-3a and TcGluCl-3c also had high expressions in the ovary and testis of T. castaneum, whereas TcGluCl-3b showed high expression in the midgut, suggesting the diverse physiological functions of TcGluCl variants in T. castaneum. The total TcGluCl and three variants showed the highest expression levels in the early stage larvae. The expressions of TcGluCl, TcGluCl-3b, and TcGluCl-3c were significantly increased from the late-stage larvae to the early stage pupae and indicated that the TcGluCl might be involved in the growth and development of T. castaneum. 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.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.