title: Strongyloides papillosus: changes in transcript levels of lysozyme and aspartic protease 2 in percutaneously migrated larvae.
abstract: The infection of the host is the crucial event in the life-cycle of parasites. To understand the molecular mechanisms of this important step, different methods are used in present studies. For analysis of changes in transcript levels the most sensitive method is the quantitative real-time PCR (qPCR). For an accurate analysis the evaluation of a set of adequate reference genes is necessary. The present study aimed to analyse the transcriptional levels of two genes of interest, the putative aspartic protease Spa-asp-2 and the putative lysozyme Spa-lys, in infective, free-living larvae of Strongyloides papillosus at different ages and from long-term and short-term infections and percutaneously migrated ("parasitic") larvae. Percutaneously migrated larvae were collected using the PERL chamber system and ovine skin in vitro. Reference genes identified as most suitable for transcriptional analysis according to geNorm analysis were genes for the eukaryotic translation elongation factor 1 alpha (Spa-eft-2), actin variation 2 (Spa-act-v2) and beta tubulin (Spa-tbb-1). Transcriptional analysis of the genes in percutaneously migrated larvae showed an upregulation of Spa-asp-2, while Spa-lys was downregulated. Data from the presented study provide a first glance into the changes of transcript levels of S. papillosus induced by percutaneous migration.
authors.affiliation: Institute for Parasitology, University of Veterinary Medicine Hannover, Bünteweg 17, 30559 Hannover, Germany.
authors.forename: Valerie
authors.initials: V
authors.lastname: Biewener
authors.forename: Claudia
authors.initials: C
authors.lastname: Welz
authors.forename: Grisada
authors.initials: G
authors.lastname: Khumpool
authors.forename: Ulla
authors.initials: U
authors.lastname: Küttler
authors.forename: Thomas
authors.initials: T
authors.lastname: Schnieder
mesh_terms: D000818:Animals; D057055:Aspartic Acid Proteases / Q000235:genetics* / Q000378:metabolism; D002417:Cattle; D002418:Cattle Diseases / Q000469:parasitology; D018076:DNA, Complementary / Q000096:biosynthesis / Q000737:chemistry; D017238:Genes, Helminth; D007814:Larva / Q000201:enzymology / Q000235:genetics; D009113:Muramidase / Q000235:genetics* / Q000378:metabolism; D018510:RNA, Helminth / Q000235:genetics / Q000302:isolation & purification; D060888:Real-Time Polymerase Chain Reaction; D012756:Sheep; D012757:Sheep Diseases / Q000469:parasitology; D012867:Skin / Q000469:parasitology; D013321:Strongyloides / Q000201:enzymology* / Q000235:genetics; D013322:Strongyloidiasis / Q000469:parasitology / Q000662:veterinary; D015854:Up-Regulation
chemical_list: D018076:DNA, Complementary; D018510:RNA, Helminth; D009113:Muramidase; D057055:Aspartic Acid Proteases