Project description:A protein pilot dataset detecting Wolbachia proteins from protein extracted from dissected infected Culex pipiens mosquito ovaries. The experiment was based of an iTRAQ experiment comparing infected and uninfected ovarian tissues and has been usefull in characterizing the wPip (Buckeye) ovarian proteome.
Project description:Arboviruses are defined by their ability to replicate in both mosquito vectors and mammalian hosts. There is good evidence that arboviruses “prime” their progeny for infection of the next host, such as differential glycosylation of their outer glycoproteins or packaging of host ribosomal subunits. We and others have previously shown that mosquito derived viruses more efficiently infect mammalian cells than mammalian derived viruses. These observations are coherent with arboviruses acquiring host-specific adaptations, and we hypothesized that virus derived from either the mammalian host or mosquito vector will elicit different responses when infecting the mammalian host. Here we perform an RNA-sequencing analysis of the transcriptional response of Human Embryonic Kidney 293 (HEK-293) cells to infection with either mosquito (Aedes albopictus, C7/10) or mammalian (Baby Hamster Kidney, BHK) derived Sindbis virus (SINV). We show that C7/10 derived virus infection leads to a more robust transcriptional response in HEK-293s as compared to infection with the BHK derived virus. Surprisingly, despite more efficient infection, we found an increase in interferon-β (IFN-β) and interferon stimulated gene (ISG) transcripts in response to C7/10 derived virus infection versus BHK derived virus infection. However, translation of interferon stimulated genes was lower in HEK-293s infected with C7/10 derived virus, starkly contrasting with the transcriptional response. This inhibition of ISG translation is reflective of a more rapid overall shut-off of host cell translation following infection with C7/10 derived virus. Finally, we show that C7/10 derived virus infection of HEK-293 cells leads to elevated levels of phosphorylated eukaryotic translation elongation factor-2 (eEF2), identifying a potential mechanism leading to the more rapid shut-off of host translation. We postulate that the rapid shut-off of host translation in mammalian cells infected with mosquito derived virus acts to counter the IFN-β stimulated transcriptional response.
Project description:We conducted a genome wide survey of mosquito gene expression profiles in 4 different tissues of adult Anopheles gambiae mosquitoes. The tissues included the head, the midgut, the carcass (corresponding to the remainder of the mosquito after decapitation and midgut removal) and ovaries. Ovaries were collected from adult female mosquitoes, which had been bloodfed 48h prior to dissection. To investigate mosquito expression in those tissues, we performed competitive two-dye hybridizations of experimental and standard reference RNA samples to MMC1 microarrays. MMC1 microarrays contain approximately 20,000 EST clones. Standard reference RNA was produced in vitro from the spotted ESTs and was utilized to provide consistent, non-zero reference values for almost all probes of the array. For the experiment, two biological replicates, corresponding to mosquito generations 1 and 3 (generation 2 was discarded due to probable labelling errors) and one technical (dye-swap) replicate was conducted.
Project description:Laodelphax striatellus is naturally infected with the Wolbachia strain wStri, which significantly increase the fecundity of its host. Wolbachia-infected females produce 30%–40% more eggs than Wolbachia-uninfected females. MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that play critical roles in the regulation of gene expression at post-transcriptional level. Here we report the differentially expressed miRNAs between Wolbachia-infected and Wolbachia-uninfected strains of L. striatellus ovaries. Our data may be helpful to explore the molecular mechanisms by which Wolbachia increase the fecundity of Laodelphax striatellus.
Project description:Female mosquitoes require a blood meal for oogenesis, and thereby receive a substantial iron load in the forms of holo-transferrin and hemoglobin. Our previous data showed that during digestion of a blood meal, the gut iron concentration decreases 10-fold, while that of the ovaries doubles from ingestion to 72 hours post feeding. Approximately 72 hours post feeding, eggs are laid with ~125 ng Fe each. We are interested in the effects of the blood meal on the expression of iron related proteins detected in the ovaries during time post feeding before eggs are laid. We have used shotgun proteomic analysis to identify proteins in the developing ovaries of Aedes aegypti; this information provides further insight into the effect of a blood meal on mosquito oogenesis.
Project description:Aedes aegypti [Linnaeus in Hasselquist (Diptera: Culicidae); yellow fever mosquito] transmits several viruses that infect millions of people each year including, Zika, dengue, yellow fever, chikungunya, and West Nile. Disease transmission occurs during blood feeding. Only the females blood feed as they require a bloodmeal for oogenesis. In the bloodmeal, females receive a substantial iron load in the forms of holo-transferrin and hemoglobin. We are interested in the effects of the iron in a bloodmeal on the expression of proteins during oogenesis. Our previous data showed that during digestion of a blood meal, the gut iron concentration decreases 10-fold, while that of the ovaries doubles from ingestion to 72 hours post feeding. Approximately 72 hours post feeding, eggs are laid with ~125 ng Fe each. We are interested in the effects of the blood meal iron on the expression of proteins detected in the ovaries during the early oogenesis, 24 hours post feeding, before eggs are laid. We have used tandem mass tag-labeling proteomics to quantify proteins expressed at this early stage following feeding of a controlled iron diet. Our findings provide the first quantitative report of differential ovary protein expression in early oogenesis in mosquitoes fed three different iron diets.
Project description:We sequenced the small RNA profiles in ZIKV-infected and non-infected Ae. aegypti mosquitoes at 2, 7 and 14 days post-infection. ZIKV induced an RNAi response in the mosquito with virus-derived short interfering RNAs dramatically increased in abundance post-infection. Further, we found 17 host miRNAs that were modulated by the ZIKV infection at all time points.
Project description:To confirm that female-to-male sexual fate reversal in Smad4flox/floxMerCreMer Stra8−/− ovaries occurs independently of somatic environment. We analyzed transcriptome of samples using RNA from control testes, ovaries, Smad4flox/floxMerCreMer Stra8+/− and Smad4flox/floxMerCreMer Stra8−/− ovaries.
Project description:The mosquito-borne chikungunya virus (CHIKV) poses a threat to human health in tropical countries throughout the world. The molecular interactions of CHIKV with its mosquito vector, Aedes aegypti are not fully understood. Following oral acquisition of CHIKV via salinemeals, we analyzed changes in the proteome of Ae. aegypti in 12 h intervals by label-free timsTOF Pro mass spectrometry. For each of the seven time points, between 2647 and 3167 proteins were identified among CHIKV infected and non-infected mosquito samples and fewer than 6% of those identified proteins were affected by the virus.