Project description:A pyrethroid-resistant strain of Culex quinquefasciatus, JPal-per, exhibits 2500-fold greater larval resistance to permethrin than the insecticide-susceptible strain Ogasawara. An increased microsome monooxygenase metabolism is involved in the resistance mechanism. Microarray analysis revealed altered expressions of cytochrome P450 genes in the fourth instar larvae of JPal-per compared to those in OGS. An oligo DNA array was designed for 62 different cDNA segments encoding unique P450 isoforms of the Cx. pipiens complex. Other probes for non-P450 Cx. pipiens complex genes were also mounted on the array.
Project description:The Southern house mosquito, Culex quinquefasciatus, is a vector of the causative agents of many diseases including West Nile Fever, St. Louis Encephalitis, and lymphatic filariasis. In order to manage the spread of these diseases, vector control efforts rely heavily on insecticides, including pyrethroids, namely permethrin. In our study we investigated the changes in the gene expression profiles of a highly-permethrin resistant strain of Cx. quinquefasciatus during constant exposure to permethrin at the LC-0, LC-50, and LC-70 rates, which killed 0, 50, and 70% of all larvae, respectively. Overall, we identified several genes that were up-regulated including detoxification genes such as cytochrome P450s as well as genes that were down-regulated.
Project description:A pyrethroid-resistant strain of Culex quinquefasciatus, JPal-per, exhibits 2500-fold greater larval resistance to permethrin than the insecticide-susceptible strain Ogasawara. An increased microsome monooxygenase metabolism is involved in the resistance mechanism. Microarray analysis revealed altered expressions of cytochrome P450 genes in the fourth instar larvae of JPal-per compared to those in OGS. An oligo DNA array was designed for 62 different cDNA segments encoding unique P450 isoforms of the Cx. pipiens complex. Other probes for non-P450 Cx. pipiens complex genes were also mounted on the array. Two strains with two color experiment, total 10 replicates, 5 biological replicates (each strain), 5 dye-swap experiments.
Project description:Culex pipiens molestus and Cx. p. quinquefasciatus are the members of Culex pipiens Complex, but they display relatively large differences in behavior and physiological responses. We compared the genes of these mosquitoes to identify those that were differentially expressed in each subspecies. Such genes could play important roles in subspecies-specific blood feeding or oviposition behavior. Culex pipiens molestus and Cx. p. quinquefasciatus females were undertaken Illumina RNA sequencing.
Project description:Culex pipiens molestus and Cx. p. quinquefasciatus are the members of Culex pipiens Complex, but they display relatively large differences in behavior and physiological responses. We compared the genes of these mosquitoes to identify those that were differentially expressed in each subspecies. Such genes could play important roles in subspecies-specific blood feeding or oviposition behavior.
Project description:Analysis of Culex quinquefasciatus responses to West Nile virus (WNV) infection at 7 and 14 days after ingestion of infected blood in the gut and carcass tissues.
Project description:The genomes of three major mosquito vectors of human diseases, including Anopheles gambiae, Aedes aegypti, and Culex pipiens quinquefasciatus, have been previously sequenced. C. p. quinquefasciatus has the largest number of predicted protein-coding genes, which partially results from the expansion of three detoxification gene families: cytochrome P450 monooxygenases (P450), glutathione S-transferases (GST), and carboxylcholinesterases (CCE). However, unlike A. gambiae and A. aegypti, which have large amounts of gene expression data, C. p. quinquefasciatus has limited transcriptomic resources. Knowledge of complete gene expression information is very important for the exploration of the functions of genes involved in specific biological processes. In the present study, the three detoxification gene families of C. p. quinquefasciatus were analyzed for phylogenetic classification and compared with those of three other dipteran insects. Gene expression during various developmental stages and the differential expression responsible for parathion resistance were profiled using the digital gene expression (DGE) technique. Results: A total of 291 detoxification genes were found in C. p. quinquefasciatus, including 70 CCE, 186 P450, and 35 GST genes. Compared with three other dipteran species, gene expansion in Culex mainly occurred in the CCE and P450 families, where the genes of M-NM-1-esterases, juvenile hormone esterases, and CYP325 of the CYP4 subfamily showed the most pronounced expansion on the genome. A total of 13314 genes were expressed in five DGE libraries. Genes with signal transduction and odorant binding functions were prominently expressed during egg development. Genes involved in proteolysis, glycosphingolipid biosynthesis, and purine metabolism were preferentially expressed at the larval stage. Seventy five percent of the detoxification genes were found to be expressed. One fourth of the CCE and P450 genes were expressed at unique stages, indicating their developmentally regulated expression. Fifteen detoxification genes, including 2 CCEs, 6 GSTs, and 7 P450s, were expressed at higher levels in a parathion-resistant strain than in a susceptible strain. Conclusion: The results of the present study provide new insights into the functions and evolution of three detoxification gene families in mosquitoes and comprehensive transcriptomic resources for C. p. quinquefasciatus, which will facilitate the elucidation of molecular mechanisms underlying the different biological characteristics of the three major mosquito vectors. Raw data were deposited in SRA and assigned accession number SRA049959: http://www.ncbi.nlm.nih.gov/sra?term=SRA049959 Five DGE libraries were sequenced: the egg, third instar larval, pupal, and adult stages of the SG strain, and the third instar larval stage of the S-lab strain.
Project description:Illumina HiSeq 2000 was conducted on the mRNA fraction of fourth instar Cx. quinquefasciatus from a highly permethrin resistant strain (HAmCqG8) and its parental low resistance strain (HAmCqG0). After aligning reads to the Johannesburg genome using the GTF annotations file from Ensembl Metazoa, we found that 367 genes were upregulated in the high resistant strain and 3982 were downregulated. The upregulated genes were associated with cytochrome P450s and proteases while the downregulated genes were mostly found in the “no annotation” category for the Structural Classification of Proteins classifications for Cx. quinquefasciatus. This study identified the upregulated genes in the highly resistant HAmCqG8 strain that may be involved in resistance to permethrin.
Project description:Illumina HiSeq 2000 was conducted on the mRNA fraction of fourth instar Cx. quinquefasciatus from a highly permethrin resistant strain (HAmCqG8) and its parental low resistance strain (HAmCqG0). After aligning reads to the Johannesburg genome using the GTF annotations file from Ensembl Metazoa, we found that 367 genes were upregulated in the high resistant strain and 3982 were downregulated. The upregulated genes were associated with cytochrome P450s and proteases while the downregulated genes were mostly found in the “no annotation” category for the Structural Classification of Proteins classifications for Cx. quinquefasciatus. This study identified the upregulated genes in the highly resistant HAmCqG8 strain that may be involved in resistance to permethrin. RNA Seq profiling of the mRNA of fourth instar HAmCqG0 and HAmCqG8 Cx. quinquefasciatus (whole insect pooled samples)