title: Dirofilaria immitis exhibits sex- and stage-specific differences in excretory/secretory miRNA and protein profiles.
abstract: The canine heartworm Dirofilaria immitis releases excretory/secretory molecules into its host and in culture. We report analyses of the types, amounts and stage-dependence of microRNAs and proteins found in D. immitis culture media recovered after incubating 800,000 microfilariae for 6days, 500L3and 500L4for 7days, as well as 40 adult females and 40 adult males for 48h, all separately. In addition, the presence of exosome-like particles was established by nanoparticle tracking analysis. Our results are in concordance with the D. immitis molecules previously detected in dog blood and in culture medium, but add additional insight into the sex- and stage-specificity of these processes. Of 131 miRNA candidates analyzed, none of the most abundant sequences was exclusively associated with one stage. Several isoforms of the nematode miR-100 family, miR-279, miR-71, were highly represented and overlapped substantially with the profile of heartworm miRNAs described from infected dog blood. lin-4 was primarily associated with males. We also report 4, 27 and 72 proteins in media from microfilariae, females and males, respectively. The only protein in common to all samples was actin, and only 9/88 proteins with a gene ontology description had not been reported in other studies of filarial secretomes. Exosomal proteins were well represented, dominated by cytoskeletal proteins, metabolic enzymes, zeta polypeptide, and chaperones.
authors.affiliation: Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, 21,111 Lakeshore Road, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada.
authors.forename: Lucienne
authors.initials: L
authors.lastname: Tritten
authors.affiliation: Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, 21,111 Lakeshore Road, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada.
authors.forename: Damian
authors.initials: D
authors.lastname: Clarke
authors.affiliation: Zoetis Inc., 333 Portage Street, Kalamazoo, MI 49008, USA.
authors.forename: Scott
authors.initials: S
authors.lastname: Timmins
authors.affiliation: Zoetis Inc., 333 Portage Street, Kalamazoo, MI 49008, USA.
authors.forename: Tom
authors.initials: T
authors.lastname: McTier
authors.affiliation: Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, 21,111 Lakeshore Road, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada. Electronic address: timothy.g.geary@mcgill.ca.
authors.forename: Timothy G
authors.initials: TG
authors.lastname: Geary
mesh_terms: D000818:Animals; D004183:Dirofilaria immitis / Q000254:growth & development / Q000502:physiology*; D004184:Dirofilariasis / Q000097:blood* / Q000469:parasitology; D004283:Dog Diseases / Q000097:blood* / Q000469:parasitology; D004285:Dogs; D005260:Female; D015801:Helminth Proteins / Q000097:blood*; D008018:Life Cycle Stages / Q000502:physiology*; D008297:Male; D035683:MicroRNAs / Q000032:analysis*; D012737:Sex Factors
chemical_list: D015801:Helminth Proteins; D035683:MicroRNAs
keywords: Dirofilaria immitis; Protein; Secretion; Secretome; Stages; microRNA