title: RNA interference mediated knockdown of the KDEL receptor and COPB2 inhibits digestion and reproduction in the parasitic copepod Lepeophtheirus salmonis.
abstract: Retrograde transport of proteins from the endoplasmic reticulum to the Golgi is an essential part of the secretory pathway that all newly synthesised secreted and membrane proteins in eukaryotic cells undergo. The aim of this study was to characterise two components of the retrograde transport pathway in the parasitic copepod Lepeophtheirus salmonis (salmon louse) on a molecular and functional level. LsKDELR and LsCOPB2 were confirmed to be the salmon louse homologues of the chosen target proteins by sequence similarity. Ontogenetic analysis by qRT-PCR revealed the highest expression levels of both genes in adult females and the earliest larval stage. LsKDELR and LsCOPB2 localisation in adult females was detected by immunofluorescence and in situ hybridisation, respectively. Both LsKDELR and LsCOPB2 were found in the ovaries, the oocytes and the gut. LsKDELR and LsCOPB2 were knocked down by RNA interference in preadult females, which was confirmed by qRT-PCR. LsCOPB2 knockdown lice had a significantly higher mortality and failed to develop normally, while both LsCOPB2 and LsKDELR knockdown caused disturbed digestion and the absence of egg strings. This shows the potential of LsKDELR and LsCOPB2 as suitable target candidates for new pest control methods.
authors.affiliation: Department of Biology, University of Bergen, P.O. Box 7803, 5020 Bergen, Norway. Electronic address: christiane.trosse@bio.uib.no.
authors.forename: Christiane
authors.initials: C
authors.lastname: Tröße
authors.affiliation: SLRC-Sea Lice Research Centre, Department of Biology, University of Bergen, P.O. Box 7803, 5020 Bergen, Norway.
authors.forename: Frank
authors.initials: F
authors.lastname: Nilsen
authors.affiliation: SLRC-Sea Lice Research Centre, Institute of Marine Research, P.O. Box 1870, 5817 Bergen, Norway.
authors.forename: Sussie
authors.initials: S
authors.lastname: Dalvin
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D020756:Coatomer Protein / Q000235:genetics* / Q000378:metabolism; D033342:Copepoda / Q000166:cytology / Q000502:physiology*; D004063:Digestion / Q000502:physiology; D055785:Gene Knockdown Techniques; D034622:RNA Interference / Q000502:physiology*; D018000:Receptors, Peptide / Q000235:genetics* / Q000378:metabolism; D012098:Reproduction / Q000502:physiology; D020317:Salmo salar / Q000469:parasitology*
chemical_list: D020756:Coatomer Protein; C064459:KDEL receptor; D018000:Receptors, Peptide
keywords: COPI beta prime; ERPR; RNAi; Retrograde transport; Sea lice