Project description:Because premating behavioral isolation typically involves the interaction between traits in males and females, we generated whole-genome expression profiles of virgin males collected at the same age and diel time as the virgin females assayed in Cassone et al. (2010). Using the same Affymetrix Anopheles/Plasmodian GeneChip platform, levels and patterns of transcript abundance were examined between lab colonies of M and S Quantitative real-time PCR (qRT-PCR) was used to examine tissue-specific expression of candidate genes, as well as to validate transcription variation using RNA samples of M and S mosquitoes collected from natural populations in Burkina Faso and Cameroon. Gas chromatography-mass spectrometry was performed to examine qualitative differences in virgin male cuticular hydrocarbon composition between forms at mid-dusk and two additional time points.
Project description:shock treatment<br>Differential gene expression between the alternate 2La arrangements was examined for heat shock treated and control fourth instar larvae at three discrete time points to examine temporal variation in thermal stress response. One week after egg hatching, 72 fourth instar larvae of each 2La inversion polmorphism were randomly selected from multiple pans and placed into individual 13x100 mm glass culture tubes containing 2 mL of water. Heat shock treatment was carried out by placing tube racks containing larvae into a 38?C water bath for 60 min. After heat shock, larvae were placed back into 27?C water until collection, which took place 0.25 h, 2 h, and 8 h later by pooling 24 individuals into a 1.5 mL microcentrifuge tube and snap-freezing them in liquid nitrogen. Samples were stored at -80 ?C until RNA isolation. Control groups were treated identically for each time point with the only difference being that they were not exposed to the 60 min heat shock but rather placed into 27?C water<br>
Project description:Semi-field translocations to examine whether gene expression differences between Anopheles gambiae M and S occupying discrete larval habitats are due to inherent molecular form divergence or transcriptional plasticity
Project description:MicroRNAs (miRNA) have alternative forms known as isomiRs, which differ from each other by a few nucleotides. Next generation sequencing platforms facilitate identification of these isomiRs and recent discoveries regarding their functional importance have increased our understandings of the regulatory complexities of the microRNAome. Observed changes in the miRNA profiles in mosquitoes infected with flaviviruses have implicated small RNAs in the interactions between viruses and their vectors. Here we analysed the isomiR profiles of both uninfected and infected blood fed Aedes aegypti mosquitoes with a major human pathogen, Dengue virus at two time points post-infection. We found noticeable changes to the isomiR expression profile in response to infection and aging. Data analysis revealed a distinct bias towards isomiR production in the mature miRNA as opposed to the star strand. Furthermore, we noticed that only in 40% of Ae. aegypti miRNAs, the most abundant reads for each particular miRNA match the exact sequence reported in the miRbase. The isomiR expression variations between an Ae. aegypti embryonic cell line (Aag2) and whole mosquitoes demonstrated a tissue-specific pattern of isomiR production. Our results illustrated a bias for certain types of isomiRs for each miRNA. The findings presented in this study also provide evidence that isomiR production is not a random phenomenon and may be important in DENV colonisation of its vector. Examination of isomiR production rate in DENV infected and non infected mosquitoes
Project description:Spermatogenesis is a highly complex developmental process, during which diploid germline stem cells are transformed into motile haploid spermatozoa. The process involves a precisely regulated action of a large number of genes, making the testes the most distinct tissue within the body. Testis transcriptome has been analysed in several groups of animals, but never systematically studied in mosquitoes. This dataset, consisting of the transcriptome of the developing testes from late larvae and early pupae of the African malaria mosquito, Anopheles gambiae, closes the existing gap.
Project description:In this study we determine the effects of silencing adiponectin receptor in A. gambiae mosquitoes after taking Plasmodium gambiae infected blood meals. dsRNA (dsAdpR and dsGluc) microinjected A. gambiae mosquitoes were fed on P. berghei-infected Swiss Webster mice. Two days after a blood meal, the mosquitoes were collected for midgut dissection.
Project description:Identification of genes associated with bendiocarb resistance. Mosquitoes collected as larvae from Nagongera and Kihihi, Uganda. Bendiocarb-resistant and unexposed female mosquitoes selected using standard WHO tube bioassays. RNA was extracted from pools of five individuals identified as An. gambiae s.s. Insecticide-susceptible mosquitoes from the Kisumu strain were included as controls. RNA hybridized in an interwoven loop design to compare four biological replicates each of resistant, unexposed, and laboratory mosquitoes.