Project description:In mammalian systems, extracellular small RNAs can operate in a paracrine manner to communicate information between cells, relying on transport within vesicles. “Foreign” small RNAs derived from bacteria, plants and parasites have also been detected in mammalian body fluids, sparking interest in whether these could mediate inter-species communication. However, there is no mechanistic framework for RNA-mediated interspecies communication and the active movement of RNA via vesicles has not been shown outside of mammals. Here we demonstrate that specific microRNAs and Y RNAs are packaged into vesicles secreted by a gastrointestinal nematode, Heligmosomoides polygyrus, which naturally infects mice. Total RNA was extracted from the secretion product of adult worms and compared to the profile of small RNAs in adult worms, eggs and infective larvae.
Project description:In mammalian systems, extracellular small RNAs can operate in a paracrine manner to communicate information between cells, relying on transport within vesicles. “Foreign” small RNAs derived from bacteria, plants and parasites have also been detected in mammalian body fluids, sparking interest in whether these could mediate inter-species communication. However, there is no mechanistic framework for RNA-mediated interspecies communication and the active movement of RNA via vesicles has not been shown outside of mammals. Here we demonstrate that specific microRNAs and Y RNAs are packaged into vesicles secreted by a gastrointestinal nematode, Heligmosomoides polygyrus, which naturally infects mice. Total RNA was extracted from the serum of mice infected with Litomosoides sigmodontis at 60 days post infection
Project description:We present a comparative analysis of sex-specific tegument proteins of paired or virgin Schistosoma mansoni. By apply a new and highly sensitive workflow, detection of even low abundance proteins from the worm was achieved Therefore, a streptavidin-biotin affinity purification technique in combination with single pot solid-phase enhanced sample preparation was established for subsequent LC-MS/MS analysis. We were able to identify 1519 tegument proteins for male and female as to virgin and paired worms. Sex-specific proteins were screened, present in both virgin and paired adult schistosomes to reduce the impact of mating. Bioinformatic analysis revealed an involvement of female-specific tegument proteins in signaling pathways of cellular processes and antioxidant mechanisms. Male-specific proteins were found to be enriched in processes linked to phosphorylation and signal transduction. This suggests a task sharing between the sexes that might be necessary for survival in the host. Our datasets provide a basis for further studies to understand and ultimately decipher the strategies of the two worm sexes to evade the immune system.
Project description:In nature, animals often face feast or famine conditions. We aimed to identify the miRNAs of Caenorhabditis elegans that changed their expression under starvation conditions in stage L4 larvae. Our results highlight 14 miRNAs that show differential expression in starved versus well-fed larvae. In particular, miRNAs of the miR-35-3p/miR-41-3p family were upregulated 6-20 fold upon starvation. We verified the upregulation of miR-35-3p with qPCR. Additionally, we showed that the expression of gld-1, important in ovogenesis, and a validated target of miR-35-3p, was downregulated when the expression of miR-35-3p was higher. This study represents a starting point for a more comprehensive understanding of the role of miRNAs during starvation in C. elegans. Illumina small RNA sequencing of starved and well-fed L4 worms.
Project description:In this study, the composition of ES of male and female L4 stage Heligmosomoides polygyrus bakeri in the presence (cultured together) or absence (cultured alone) of the opposite sex was examined using mass spectrometry.
Project description:Schistosoma mansoni is the major causative agent of schistosomiasis in the Americas. This parasite takes advantage from host signaling molecules such as cytokines and hormones to complete its development inside the host. TNF-α is the most important cytokine involved in the inflammatory response when cercaria, the infective stage, penetrates the human skin and a severe inflammatory response is started. In this work the authors describe the complete sequence of a possible TNF-α receptor in S. mansoni and detect that the receptor is most highly expressed in cercaria among all life cycle stages. In an attempt to mimic the situation at the site of skin penetration cercariae have been mechanically transformed in vitro into schistosomula and immediately exposed to human TNF-α . Exposure of these early schistosomula to the human hormone caused a large-scale change in the expression of parasite genes. Exposure of adult worms to human TNF-α caused gene expression changes as well, although the set of parasite altered genes was entirely different from that of schistosomula. This work increases the number of known signaling pathways of the parasite, and opens new perspectives into understanding the molecular components of TNF-α response as well as possibly interfering with parasite-host interaction. A platform previously designed by our group (PMID: 17517391) was used. 300 ng of RNA from adult schistosomes treated with TNF and its control were used for the hybridization. RNA amplification and labeling kit (Agilent Technologies) were used according to manufacturer´s instructions. Each sample was separately labeled with either Cy3 or Cy5. 825 ng cRNA from each amplification were used for hybridization; self-self experiments were performed. Washing and scanning procedures were according to the manufacturer´s instructions using GenePix 4000B scanner (Molecular Devices, Sunnyvale, CA, USA). Data was extracted using Feature Extraction software (Agilent Technologies). We calculated a virtual Log2 ratio between intensities of TNF-α Treated/Control, for each gene. With these ratios, we used two different approaches for SAM statistical analyses. SAM one-class approach was used to identify genes with sustained changes in their expression levels along the entire time period of observation (1 and 24 h); SAM two-class approach was used to identify genes with transient changes in their expression levels. In both cases, genes were considered differentially expressed at q-value � 0.05. Hierarchical clustering of selected genes was generated using Spotfire Decision Site software (TIBCO Software Inc., Palo Alto, CA, USA). For a gene that was represented in the array by multiple probes, we picked a single representative probe by selecting the probe with the highest absolute value of the Log2 ratio between intensities of TNF-α Treated/Control.
Project description:Ectoparasitic flatworms from the family Diplozoidae (Platyhelminthes: Monogenea) represent serious hematophagous fish pathogens. Information related to the biochemical and molecular nature of the physiological processes is rather sporadic, as well as the knowledge of the molecules produced by monogeneans and their role in host-parasite interaction. Therefore, we performed a complex secretomic analysis of monogenean representative Eudiplozoon nipponicum for the purpose to identify functionally important protein molecules involved in these host-parasite interactions.
Project description:The aim of this study was to investigate the response in gene expression before and after exposure to the Benzimidazoles drug flubendazole in adult female Ascaridia galli worms. The nematode Ascaridia galli (order Ascaridida) is an economically important intestinal parasite responsible for increased food consumption, reduced performance, and mortality in commercial poultry production. Parasite control relies on repeated use of dewormers (anthelmintics). Benzimidazoles are currently the only anthelmintic registered against A. galli in the EU and there is an obvious risk that overuse of one drug class may lead to resistance. The worms were collected from a commercial laying hen farms before and on day three during a treatment period of 7 days with flubendazole.
Project description:Sperm competition theory predicts that males should tailor ejaculates according to their social status. Here we test this in a model vertebrate, the house mouse (Mus musculus domesticus), combining experimental data with a quantitative proteomics analysis of seminal fluid composition. Our analyses reveal that both sperm production and the relative production of proteins found in seminal fluid differ according to social dominance. Notably, whereas dominant males produce and ejaculate more sperm, subordinate males produce greater relative amounts of key proteins used in the formation of copulatory plugs. These findings have important implications for understanding the dynamics and outcome of sperm competition.
Project description:Anoplocephala perfoliata is a common tapeworm in horse with an ability to cause colic and increase mortality. At present, current diagnostic methods are not sensitive enough to detect A. perfoliata and immunoreactive excretory/secretory (E/S) proteins produced by this parasite, which comprise the most promising candidates for diagnostic tests. Here, we compared the E/S proteins produced by A. perfoliata worms of two different physiological states (small- and large-sized worms) in vitro by label-free quantitative (LFQ) proteomics with protein database composed of proteins encoded by Hymenolepis diminuta, Echinococcus multilocularis/granulosus and Taenia aseatica for protein identifications. The quantitative proteomics data was complemented with one-dimensional gel electrophoresis combined with immunoblotting using antisera from horses diagnosed positive and negative for this tapeworm. Protein bands cross-reacting most efficiently with the anti-A. perfoliata antibodies were subjected to in-gel digestion with LC-MS/MS identification. Both the non-gel and gel-based proteomic results were compared to propose a list of candidate proteins for further studies. These proteins will be validated for their suitability as diagnostic markers specific to cestodes and A. perfoliata.