Project description:We report the RNA-seq based analyses of the transcriptional changes in the Aedes aegypti transcriptome 4 days after blood feeding with 2 μM hsa-miR-150-5p antagomir or mock.
Project description:The Anopheles gambiae midgut harbors bacteria that proliferate upon a blood feed. We used microarrays to examine the midgut gene expression response at early stages (3hours) after an artifitial meal containing heat killed bacteria. Anopheles gambiae G3 mosquitoes 5-6 day-old were fed BSA (20% in PBS with fresh 10 mM sodium bicarbonate) with or without heat killed E. coli (equivalent of 2.5 ml of 0.8 OD) . Three pools of 10 mosquito midguts were dissected after 3h and processed for microarray analysis of gene expression.
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:To understand the the effect of antagomir-17 treatment on human endothelial cells derived from human umbilical cord blood (UCB) CD34+ hematopoietic stem cells, we have employed mRNA sequencing. The antagomiR-17 used in this study was purchased from Dharmacon and cell transfection was performed using Lipofectamine RNAiMAx from Life Technologies. Scramble antagomiR from Ambion was used as control. Cells were transfected with antagomiR-17 or scrambled antagomiR for 48 hours. After 48 h, the cells were collected, RNA was isolated and RNA samples were shipped to Exiqon Services, Denmark for mRNA sequencing. All sequencing experiments (RNA integrity measurements, library preparation and next generation sequencing) were conducted at Exiqon Services, Denmark.
Project description:Fat body is an important tissue in the context of vitellogenesis, vector immunity, vector physiology and vector-parasite interaction. However, the proteome of fatbody and impact of blood meal on the gene expression of this vital organ has not been investigated so far. Therefore, in this study, we made an attempt to identify proteins expressed in fatbody of An. stephensi and their altered expression in response to blood meal. In all, we identified 4,504 proteins in the fatbody using multiple fractionation strategies, which is by far the largest resource of fatbody proteome in any mosquito species. Further, comparative proteomic analysis of fatbody 24 and 48 hours post blood meal led to identification of over 300 differentially expressed proteins. Bioinformatics analysis of these proteins suggested their role in vitellogenesis, lipid transport, mosquito immunity and oxidation-reduction processes. Interestingly, we identified four novel genes,which were found to be differentially expressed upon blood meal. These proteins are potential target for vector control strategies and development of transmission blocking vaccines.
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:The Anopheles gambiae midgut harbors bacteria that proliferate upon a blood feed. We used microarrays to examine the midgut gene expression response at early stages (3hours) after an artifitial meal containing heat killed bacteria.
Project description:Aedes aegypti mosquito ovarian follicles develop synchronously after taking a blood meal. A single layer of follicular epithelial cells surrounding the oocyte is responsible for secreting a majority of eggshell structural components between 18 and 54 hours post blood meal. We previously identified a protein called eggshell organizing factor 1 (EOF1). When we knocked down EOF1 with RNA interference in female adult mosquitoes, they laid eggs that were not properly melanized, and fragile eggs did not contain viable embryos. Here, we aimed to determine putative downstream eggshell proteins in RNAi-EOF1 female mosquitoes. Our eggshell proteomics identified 220 proteins, suggesting that mosquito extracellular eggshells are composed of a complex mixture of proteins. These proteins are involved in the formation of an intact eggshell that protects the embryonic development and larva from the environment for long periods of time.
Project description:Plasmodium sporozoites are injected, in addition to saliva, into animal hosts when a female Anopheles mosquito takes a blood meal. The molecular components of saliva that interact with Plasmodium during this process are poorly characterized. Here we collected Plasmodium sporozoites directly from salivating Anopheles mosquitoes and looked for the presence of vector proteins that could be interacting with the parasites during transmission for further characterization.