Project description:Brugia malayi is a parasitic nematode that causes lymphatic filariasis in humans. A total of 178 novel microRNA were identified from short read transcriptional data, which when combined with known Brugia microRNAs yielded a total of 284 microRNA. Of these, 123 microRNA sequences (43%) are differentially expressed over the mammalian life stages of B. malayi that we examined. Putative targets of these microRNA were identified from inversely expressed target clusters that contain valid seed sequences for the corresponding microRNAs. The largest identified cluster is downregulated in adult females and enriched in zinc finger domains, helicase domains, and DNA binding domains suggesting this microRNA cluster may have regulatory control over a large proportion of adult female specific mRNA genes. MicroRNA-like molecules are identified as produced by the Wolbachia endosymbiont, providing evidence for direct nucleic acid-based interdomain communication between filarial nematodes and their bacterial obligate endosymbiont.
Project description:Brugia pahangi is a parasitic nematode that is closely related to B. malayi and Wuchereria bancrofti. B. malayi and W. bancrofti are responsible for lymphatic filariasis, affecting around 120 million people in 73 countries worldwide.This project aims to undertake high-throughput sequencing of Brugia pahangi transcriptome. The objective is to use transcriptomics to support gene finding and to recognize genes expressed in given life stages.
Project description:Comparative analysis of gene expression profiles provided novel insights into the genes that are transcriptionally active in infective and developing larvae of two closely related species. Species differences may indicate different metabolic adaptations that could affect host specificity, tissue tropism, and pathogenicity Two biological replicates of infective (L3) or developing larval RNA used for hybridization, in duplicate, to examine the gene expression changes in Brugia larvae Brugia malayi vector derived third stage larvae (Bm VL3); Brugia pahangi vector derived third stage larvae (Bp VL3); Brugia pahangi L3 cultured in vitro (Bp cL3); Brugia pahangi L3 derived from peritoneal cavity of infected gerbils (Bp ipL3); Brugia pahangi migrating L3 (Bp mL3) from infected gerbils
Project description:Lymphatic filarial nematodes maintain a mutualistic association with the intracellular bacterium Wolbachia. Wolbachia populations expand following infection of the mammalian host, to support larval growth and development. Utilizing transcriptomic data from Brugia malayi over the first two weeks post-infection, we present an analysis of the biochemical pathways that are involved in Wolbachia population growth and regulation in support of larval development. In Wolbachia, we observe coordinated regulation of carbon metabolism with an alternating pattern of glycolysis and TCA cycle pathways reminiscent of the ‘Warburg effect’. Wolbachia's purine, pyrimidine and haem biosynthesis and Type IV secretion pathways are also upregulated and correlate with the upregulation of the nematode’s DNA replication pathway. In the nematode we observe up-regulation of the autophagy pathway, a key regulator of Wolbachia populations. These findings support a key role for nucleotide and haem provisioning from Wolbachia in support of the larval growth and development of its nematode host.