Seminal fluid and sperm proteomics of Aedes aegypti
ABSTRACT: Sperm samples were extracted from adult A. aegypti male seminal vesicles. Semen samples were extracted from bursa of recently mated bursa of N15 labeled females. The goal was to differentiate between the contribution of seminal fluid proteins and sperm proteins in the semen transfered to the female. Our results yield insights into the molecular function, genome organization, regulation, and evolution of sperm proteins and SFPs in this important disease vector.
The yellow fever mosquito, Aedes aegypti,, transmits several viruses causative of serious diseases, including dengue, Zika, and chikungunya. Some proposed efforts to control this vector involve manipulating reproduction to suppress wild populations or to replace them with disease-resistant mosquitoes. The design of such strategies requires an intimate knowledge of reproductive processes, yet our basic understanding of reproductive genetics in this vector remains largely incomplete. To accelerate ...[more]
Project description:Investigation of whole genome gene expression level changes of testes in the meiotic drive system in aedes aegypti during spermatogenesis compared to non drive strain. The meiotic drive system in Aedes aegypti causes the female determining chromosome to fragment during spermatogenesis. A six chip study using total RNA from three separately extracted non driving strain testes of Aedes aegypti and three separately extracted meiotic drive strain testes of Aedes aegypti.
Project description:Aedes aegypti is a major vector for dengue, chikungunya and yellow fever. Though salivary gland plays a crucial role in transmission of virus and vector life cycle, there is no global and unbiased published report on salivary gland proteome of Ae. aegypti. In this study, we have carried out mass spectrometry based proteomic analysis of salivary gland from adult female Ae. aegypti mosquitoes. By fractionating the proteins isolated from salivary gland on SDS-PAGE and analyzing the in-gel digested bands on high resolution mass spectrometry, we identified 1,205 proteins. This is by far the largest catalogue of salivary gland proteome of Ae. aegypti. These proteins were then assigned molecular functions and biological processes. Several immunity related pathways were found to be enriched in salivary gland. In addition, subset of proteins was predicted to secretory in nature and may play an important role during blood feeding. This study provides a useful resource of proteins expressed in salivary gland of Ae. aegypti female mosquitoes and will aid in biomedical research focused on development of transmission blocking vaccine.
Project description:Bacillus thuringiensis israelensis (Bti) toxins are increasingly used for mosquito control, but little is known about the precise mode of action of each of these toxins, and how they interact to kill mosquito larvae. By using RNA sequencing, we investigated change in gene transcription level and polymorphismvariations associatedwith resistance to each Bti Cry toxin and to the full Bti toxin mixture in the dengue vector Aedes aegypti. The upregulation of genes related to chitin metabolismin all selected strain suggests a generalist, non-toxin-specific response to Bti selection in Aedes aegypti. Changes in the transcription level and/or protein sequences of several putative Cry toxin receptors (APNs, ALPs, α-amylases, glucoside hydrolases, ABC transporters) were specific to each Cry toxin. Selective sweeps associated with Cry4Aa resistancewere detected in 2 ALP and 1 APNgenes. The lack of selection of toxin-specific receptors in the Bti-selected strain supports the hypothesis that Cyt toxin acts as a receptor for Cry toxins in mosquitoes.
Project description:Investigation of whole genome gene expression level changes during a 44 hour time period in Aedes aegypti observed under light/dark and constant dark conditions. This is a 12-plex high defination NimbleGen array design. The Higgs White Eye (Wh) strain of Aedes aegypti was investigated for rhythmic expression during the light/dark phase and constant darkness. Samples were collected every 4 hours for 44 hours in each treatment, and RNA collected from whole head tissue.
Project description:Whole genome transcriptional profiling comparing Ae. aegypti infected with Wolbachia sp. wMelPop (PGYP1) to an uninfected Ae. aegypti control line (PGYP1.tet). The objective of the experiment was to identify genes that may be involved in the life shortening phenotype associated with wMelPop infection. Two-colour experiment; infected vs. uninfected Ae. aegypti; 4 biological replicates; 2 dye swaps.
Project description:The incomplete genome annotation of non-model organisms hampers molecular and proteomic studies. Proteomics informed by transcriptomics (PIT) is suited to non-model organisms because peptides are identified using transcriptomic, not genomic, data. Aedes aegypti is the mosquito vector for the (re-)emerging dengue, chikungunya, yellow fever and Zika viruses. An Ae. aegypti genome sequence is available, however experimental evidence for >90% of the Ae. aegypti proteome or the activity of transposable elements (TEs) that constitute 50% of the Ae. aegypti genome is lacking. We used PIT to characterise the proteome of the Aedes aegypti derived cell line Aag2. Hotspots of incomplete genome annotation were identified which are not explained by poor sequence and assembly quality. We developed criteria for the characterisation of proteomically active TEs and demonstrate that protein expression does not correlate with a TE’s genomic abundance. Finally, we identify Phasi Charoen-like virus as an unrecognised contaminant of Aag2 cells. We therefore present the first proteomic characterisation of mobile genetic elements, and provide proof-of-principle that PIT can evaluate a genome’s annotation to guide annotation efforts.
Project description:Microarray analysis on days 1, 2 and 7 post-infection with dengue, yellow fever and West Nile virus in Aedes aegypti Rockefeller strain mosquitoes RNA was purified and hybridized with Nimblegen X4 microarray chips using 81-mer probes designed from 18,000 open reading frames (ORF) found in the Ae. aegypti genome, with 2 different probes per ORF Nimblegen X4 array format, 81mer probes, RNA samples taken mostly in triplicate, microarray analysis done in triplicate. Thirty seven samples in total. Fold change data with infection/mock on the Series record.
Project description:We reported the RNA-seq based analyses of the transcriptional changes in the Aedes aegypti midguts knock down 3, 6days, feed antibody 18h transcriptome. Comparison of the midguts transcriptome of Aedes aegypti females at two knockdown time points and one feed condition; GFP dsRNA-3 or -6days: 3 or 6 days after 7day-old mosquitos were microinjected GFP dsRNA AaMesh dsRNA-3 or -6days: 3 or 6 days after 7day-old mosquitos were microinjected AaMesh dsRNA Pre-immune-18h: 18hrs after 7day-old wild type mosquitos were fed with Pre-immune AaMesh antibody-18h: 18hrs after 7day-old wild type mosquitos were fed with AaMesh antibody
Project description:Custom microarrays were used to examine global differences in female vs. male gene expression in the developing pupal head of the dengue vector mosquito Aedes aegypti. RNA was extracted from the heads of male and female 24 hr pupae. 20 male or female heads were pooled for each of four replicates. Hybridization experiments were performed on the Nimblegen Aedes aegypti 12-plex microarray design: 090305_Aedes_aegypti_TEfam_expr.ndf. Four unique replicates and two repeat replicates were assessed in the hybridization experiment.