Multiple Cytochrome P450 Genes: Their Constitutive Overexpression and Permethrin Induction in Insecticide Resistant Mosquitoes, Culex quinquefasciatus

Four cytochrome P450 cDNAs, CYP6AA7, CYP9J40, CYP9J34, and CYP9M10, were isolated from mosquitoes, Culex quinquefasciatus. The P450 gene expression and induction by permethrin were compared for three different mosquito populations bearing different resistance phenotypes, ranging from susceptible (S-Lab), through intermediate (HAmCqG0, the field parental population) to highly resistant (HAmCqG8, the 8th generation of permethrin selected offspring of HAmCqG0). A strong correlation was found for P450 gene expression with the levels of resistance and following permethrin selection at the larval stage of mosquitoes, with the highest expression levels identified in HAmCqG8, suggesting the importance of CYP6AA7, CYP9J40, CYP9J34, and CYP9M10 in the permethrin resistance of larva mosquitoes. Only CYP6AA7 showed a significant overexpression in HAmCqG8 adult mosquitoes. Other P450 genes had similar expression levels among the mosquito populations tested, suggesting different P450 genes may be involved in the response to insecticide pressure in different developmental stages. The expression of CYP6AA7, CYP9J34, and CYP9M10 was further induced by permethrin in resistant mosquitoes. Taken together, these results indicate that multiple P450 genes are up-regulated in insecticide resistant mosquitoes through both constitutive overexpression and induction mechanisms, thus increasing the overall expression levels of P450 genes.

[1]  Fang Zhu,et al.  Co-up-regulation of three P450 genes in response to permethrin exposure in permethrin resistant house flies, Musca domestica , 2008, BMC Physiology.

[2]  N. Liu,et al.  Phenobarbital induction of CYP6D1 is due to a trans acting factor on autosome 2 in house flies, Musca domestica , 1997, Insect molecular biology.

[3]  Shinji Kasai,et al.  Overexpression of cytochrome P450 genes in pyrethroid-resistant Culex quinquefasciatus. , 2010, Insect biochemistry and molecular biology.

[4]  Ernest Hodgson,et al.  Insect Cytochrome P450 , 1991 .

[5]  L. C. Terriere.  Enzyme Induction, Gene Amplification and Insect Resistance to Insecticides , 1983 .

[6]  R. Feyereisen,et al.  Expression of the Cytochrome P450 GeneCYP6A1in the Housefly,Musca domestica , 1992 .

[7]  Z. Dvořák,et al.  Xenobiotic-induced transcriptional regulation of xenobiotic metabolizing enzymes of the cytochrome P450 superfamily in human extrahepatic tissues. , 2008, Current drug metabolism.

[8]  D. Nebert,et al.  Evolution of the P450 gene superfamily: animal-plant 'warfare', molecular drive and human genetic differences in drug oxidation. , 1990, Trends in genetics : TIG.

[9]  S. Kasai,et al.  Use of isogenic strains indicates CYP9M10 is linked to permethrin resistance in Culex pipiens quinquefasciatus , 2010, Insect molecular biology.

[10]  R. Feyereisen,et al.  4.1 – Insect Cytochrome P450 , 2005 .

[11]  Nannan Liu,et al.  CYP4AB1, CYP4AB2, and Gp-9 gene overexpression associated with workers of the red imported fire ant, Solenopsis invicta Buren. , 2004, Gene.

[12]  Tetsuo Saito,et al.  Pest Resistance to Pesticides , 1984, Springer US.

[13]  L. C. Terriere.  Induction of detoxication enzymes in insects. , 1984, Annual review of entomology.

[14]  M. Berenbaum,et al.  Cytochrome P450 and actin genes expressed in Helicoverpa zea and Helicoverpa armigera: paralogy/orthology identification, gene conversion and evolution. , 2002, Insect biochemistry and molecular biology.

[15]  J. G. Scott,et al.  Increased transcription of CYP6D1 causes cytochrome P450-mediated insecticide resistance in house fly. , 1998, Insect biochemistry and molecular biology.

[16]  C. Mullin,et al.  Molecular mechanisms of insecticide resistance : diversity among insects , 1992 .

[17]  M. Schuler.  The Role of Cytochrome P450 Monooxygenases in Plant-Insect Interactions , 1996, Plant physiology.

[18]  S. Kasai,et al.  Molecular cloning, nucleotide sequence and gene expression of a cytochrome P450 (CYP6F1) from the pyrethroid-resistant mosquito, Culex quinquefasciatus Say. , 2000, Insect biochemistry and molecular biology.

[19]  Nannan Liu,et al.  Characterization of two novel cytochrome P450 genes in insecticide‐resistant house‐flies , 2008, Insect molecular biology.

[20]  E. Cupp,et al.  Insecticide Resistance and Cross-Resistance in Alabama and Florida Strains of Culex quinquefaciatus , 2004 .

[21]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[22]  J. G. Scott,et al.  CYP6D1 protects thoracic ganglia of houseflies from the neurotoxic insecticide cypermethrin. , 1998, Archives of insect biochemistry and physiology.

[23]  Nannan Liu,et al.  Differential expression of genes in pyrethroid resistant and susceptible mosquitoes, Culex quinquefasciatus (S.). , 2007, Gene.

[24]  A. Okey.  Enzyme induction in the cytochrome P-450 system. , 1990, Pharmacology & therapeutics.

[25]  Ting Li,et al.  Inheritance of Permethrin Resistance in Culex quinquefasciatus , 2010, Journal of medical entomology.

[26]  R. Feyereisen,et al.  Constitutive overexpression of the cytochrome P450 gene CYP6A1 in a house fly strain with metabolic resistance to insecticides. , 1994, Insect biochemistry and molecular biology.

[27]  Fang Zhu,et al.  Differential expression of CYP6A5 and CYP6A5v2 in pyrethroid-resistant house flies, Musca domestica. , 2008, Archives of insect biochemistry and physiology.

[28]  J. Aerts,et al.  Selection of appropriate control genes to assess expression of tumor antigens using real-time RT-PCR. , 2004, BioTechniques.

[29]  S. R. Palli,et al.  A brain-specific cytochrome P450 responsible for the majority of deltamethrin resistance in the QTC279 strain of Tribolium castaneum , 2010, Proceedings of the National Academy of Sciences.

[30]  D. Janzen,et al.  Herbivores: Their Interaction With Secondary Plant Metabolites , 1982 .

[31]  I. Podglajen,et al.  Modes and Modulations of Antibiotic Resistance Gene Expression , 2007, Clinical Microbiology Reviews.

[32]  Z. Wen,et al.  Insect cytochromes P450: diversity, insecticide resistance and tolerance to plant toxins. , 1998, Comparative biochemistry and physiology. Part C, Pharmacology, toxicology & endocrinology.

[33]  J. G. Scott,et al.  Cytochromes P450 and insecticide resistance. , 1999, Insect biochemistry and molecular biology.

[34]  M. Berenbaum,et al.  Metabolism of linear and angular furanocoumarins by Papilio polyxenes CYP6B1 co-expressed with NADPH cytochrome P450 reductase. , 2003, Insect biochemistry and molecular biology.

[35]  J. K. Nayar,et al.  Aedes albopictus (Diptera: Culicidae): an experimental and natural host of Dirofilaria immitis (Filarioidea: Onchocercidae) in Florida, U.S.A. , 1999, Journal of medical entomology.

[36]  C. Wittwer,et al.  Continuous fluorescence monitoring of rapid cycle DNA amplification. , 1997, BioTechniques.

[37]  Kostas Iatrou,et al.  comprehensive molecular insect science , 2004 .

[38]  W. Muir,et al.  Expression of Cyp6g1 and Cyp12d1 in DDT resistant and susceptible strains of Drosophila melanogaster , 2005, Insect molecular biology.

[39]  Nannan Liu,et al.  Resistance in the mosquito, Culex quinquefasciatus, and possible mechanisms for resistance. , 2005, Pest management science.

[40]  May R. Berenbaum,et al.  Jasmonate and salicylate induce expression of herbivore cytochrome P450 genes , 2002, Nature.

[41]  S. Kasai,et al.  Genomic structures of Cyp9m10 in pyrethroid resistant and susceptible strains of Culex quinquefasciatus. , 2010, Insect biochemistry and molecular biology.