Project description:In this project we aimed to establish a proteome-based pipeline for a non-model Baikal endemic amphipod species Eulimnogammarus cyaneus (Dybowsky, 1874). We used next-generation transcriptome sequencing to create a database for protein identification and investigated the proteome of E. cyaneus with LC-MS with particular emphasis on sexual dimorphism and response to acute thermal stress. We were able to characterize ca. 1000 protein groups and found male- and female-specific proteins, as well as proteins specific for thermal stress conditions.
Project description:Over the last 20 years, the advances in sequencing technologies highlighted the unique composition of the salivary glands of blood-feeding arthropods. Further biochemical and structural data demonstrated that salivary proteins can disrupt host hemostasis, inflammation and immune response in addition to favor pathogen transmission. Previously, a Sanger-based sialome of the adult Ochlerotatus. triseriatus female salivary glands was published based on 731 ESTs. Here we revisited O. triseriatus salivary glands contents using an Illumina-sequencing approach of both male and female tissues. In the current data set we report 10,317 coding DNA sequences classified into several functional classes. The translated transcripts also served as reference database for the proteomic analysis of O. triseriatus female saliva, in which unique peptides of 101 proteins were found. Finally, comparison of male and female libraries allowed the identification of female-enriched transcripts that are potentially related to blood acquisition and virus transmission.
Project description:Simultaneous analysis of Plasmodium falciparum and Anopheles gambiae gene expression: an assessment of parasite maturation and of the mosquito response to infection. Malaria is probably the most lethal and most prevalent parasitic disease in the world, causing more than one million deaths per year. In spite of intensive research, no vaccine exists and good vaccine candidates are still a necessity. Plasmodium sporozoites from mosquito midguts are not as infective as those from salivary glands, presenting a different type of motility which is presumably associated with their invasive capacity. It is assumed that these differences are due to maturation of sporozoites, which happens either in the salivary glands or somewhere along the way from the midgut to the glands. Indeed, the invasion of the salivary gland is considered to be an essential step for malaria transmission to the host and it is known to causes expression of multiple genes in the parasite and activation of some immune-response genes in the insect. Here we compared the gene expression profile of P. falciparum parasites from midgut and salivary glands in order to search for genes involved in parasite maturation; on the assumption that any differentially expressed genes found under this condition could be potentially related to the higher infectivity of the salivary gland derived sporozoites and therefore would be potentially good vaccine candidates. Also, because very little is known about mosquito genes involved in the response to Plasmodium, we compared the gene expression profile of infected and uninfected midguts and salivary glands. Our results demonstrate that, upon infection, at least 13 genes are differentially expressed in the mosquito midgut and 43 in the salivary gland. Gene ontology enrichment analysis showed that among other things, in mosquito salivary glands, infection causes multiple genes related to chitin metabolism to be differentially expressed. Also and more importantly, at least 54 genes were differentially expressed in P. falciparum from salivary glands in comparison to midgut (19 down-regulated and 35 up-regulated). Last, we show for the first time that P. falciparum causes multiple genes in the salivary gland to be regulated and that some of those genes could be potentially beneficial to the parasite.