Tritten Lucienne, Clarke Damian, Timmins Scott, McTier Tom, Geary Timothy G
Institute of Parasitology and Centre for Host-Parasite Interactions, McGill University, 21,111 Lakeshore Road, Sainte-Anne-de-Bellevue, QC, H9X 3V9, Canada.
Zoetis Inc., 333 Portage Street, Kalamazoo, MI 49008, USA.
Vet Parasitol. 2016 Dec 15;232:1-7. doi: 10.1016/j.vetpar.2016.11.005. Epub 2016 Nov 9.
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, 500L and 500L for 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.
犬心丝虫（恶丝虫）会向其宿主以及培养环境中释放排泄/分泌分子。我们报告了对相关分子的分析结果，这些分子包括微小RNA和蛋白质，分别来自80万条微丝蚴培养6天、500条幼虫培养7天、500条幼虫培养7天以及40条成年雌虫和40条成年雄虫培养48小时后所收集的恶丝虫培养基，所有培养均单独进行。此外，通过纳米颗粒追踪分析确定了类似外泌体颗粒的存在。我们的结果与先前在犬血液和培养基中检测到的恶丝虫分子一致，但进一步深入了解了这些过程的性别和阶段特异性。在分析的131个微小RNA候选物中，最丰富的序列均未仅与某一个阶段相关。线虫miR - 100家族、miR - 279、miR - 71的几种异构体高度表达，并且与感染犬血液中描述的犬心丝虫微小RNA谱有很大重叠。lin - 4主要与雄虫相关。我们还分别报告了微丝蚴、雌虫和雄虫培养基中的4种、27种和72种蛋白质。所有样本中唯一共有的蛋白质是肌动蛋白，在丝状线虫分泌蛋白组的其他研究中，仅有9/88个具有基因本体描述的蛋白质未被报道。外泌体蛋白占比很高，主要包括细胞骨架蛋白、代谢酶、ζ多肽和伴侣蛋白。
