Project description:The Drosophila phagocytic receptor Eater is expressed specifically in phagocytic hemocytes. It contributes to host immune defense and is required for survival of bacterial infections. Eater is involved in recognition and phagocytosis of bacteria. We used microarrays to determine whether any gene expression changes after bacterial phagocytosis are dependent on the expression of the phagocytic scavenger receptor Eater. We found transcriptional regulation in response to bacterial internalization, but no significant differences between controls and samples in which eater expression had been diminshed by RNAi eater knock down.
Project description:The Drosophila phagocytic receptor Eater is expressed specifically in phagocytic hemocytes. It contributes to host immune defense and is required for survival of bacterial infections. Eater is involved in recognition and phagocytosis of bacteria. We used microarrays to determine whether any gene expression changes after bacterial phagocytosis are dependent on the expression of the phagocytic scavenger receptor Eater. We found transcriptional regulation in response to bacterial internalization, but no significant differences between controls and samples in which eater expression had been diminshed by RNAi eater knock down. Drosophila S2 cells, a hemocyte-derived cell line with phagocytic properties, were exposed to a mixture of Gram-positive and Gram-negative bacteria at conditions of 50% of cell binding. At different time points of synchronized phagocytosis (30, 90 and 180 minutes) total RNA was extracted and subjected to microarray analysis. We compared S2 cells in which Eater expression was decreased by RNA interference with control S2 cells exposed to irrelevant double-stranded (pBR322). Three independent experiments were performed corresponding to three biological replicates. Decreased phagocytosis due to RNAi eater knock down was controlled in all cases.
Project description:Background The number of managed honey bee colonies has considerably decreased in many developed countries in recent years and the ectoparasitic mites are considered as major threats to honey bee colonies and health. However, their general biology remains poorly understood. Results We sequenced the genome and transcriptomes of Tropilaelaps mercedesae, the prevalent ectoparasitic mite infesting honey bees in Asia. The de novo assembled genome sequence (353 Mb) represents 53% of the estimated genome size because of the compression of repetitive sequences; nevertheless, we predicted 15,190 protein-coding genes which were well supported by the mite transcriptomes and proteomic dataes. Although amino acid substitutions have been accelerated within the conserved core genes in of two mites, T. mercedesae and Metaseiulus occidentalis, T. mercedesae has undergone the least gene family expansion and contraction between the seven arthropods we tested. The number of sensory system genes has been dramatically reduced; meanwhile, T. mercedesae may have evolved a specialized cuticle and water homeostasis mechanisms, as well as epigenetic control of gene expression for ploidy compensation between males and females., and water homeostasis. T. mercedesae contains all gene sets required to detoxify xenobiotics, enabling it to be miticide resistant. T. mercedesae is closely associated with a symbiotic bacteriuma (Rickettsiella grylli-like) and DWVdeformed wing virus (DWV), the most prevalent honey bee virus. The presence of DWV in both adult male and female mites was also confirmed by the proteomic analysis. Conclusions T. mercedesae has a very specialized life history and habitat as the ectoparasitic mite strictly dependsing on the honey bee inside the a stable colony. Thus, comparison of the genome and transcriptome sequences with those of a tick and free-living mites and tick has revealed the specific features of the genome shaped by interaction with the honey bee and colony environment. T. mercedesae, as well as Varroa destructor, genome and transcriptome sequences not only provide insights into the mite biology, but may also help to develop measures to control the most serious pests of the honey bee.