Functional characterisation of the maternal yolk-associated protein (LsYAP) utilising systemic RNA interference in the salmon louse (Lepeophtheirus salmonis) (Crustacea: Copepoda)

Dalvin, Sussie; Frost, Petter; Biering, Eirik; Hamre, Lars Are; Eichner, Christiane; Krossøy, Bjørn; Nilsen, Frank

doi:10.1016/j.ijpara.2009.04.004

Cited by 58 publications

(75 citation statements)

References 25 publications

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“…During vitellogenesis, yolk proteins are incorporated into the oocytes (Dalvin et al, 2009;Dalvin et al, 2011;Eichner et al, 2008). We found the expression of the yolk and yolk associated proteins LsVit1, LsVit2 and LsYAP to be decreased in both LsKDELR and LsCOPB2 knock down lice when compared to the control group (Fig.…”

Section: Discussionmentioning

confidence: 72%

“…6B shows a corresponding section of a control louse with intact vitellogenic oocytes. There were also changes in the subcuticular tissue where yolk proteins are produced (Dalvin et al, 2009) (Fig. 6C).…”

Section: Functional Analysis Of Lskdelr and Lscopb2mentioning

confidence: 95%

“…LsKDELR protein was detected in the ovaries, situated in the anterior of the cephalothorax of the salmon louse, in the oviduct, bridging over to the genital segment, and in the oocytes that fill up the genital segment tightly packed in the long, coiled egg string structure typical for the salmon louse. The subcuticular tissue, which is the tissue localised below the cuticle in the cephalothorax (Dalvin et al, 2009), and the epithelium of the gut, a place of active secretion of digestive enzymes, were also positive (Fig. 2).…”

Section: Localisation Of Lskdelr and Lscopb2mentioning

confidence: 99%

“…The RNAi experiment was carried out as previously described by Dalvin et al (2009). Briefly, dsRNA fragments with a length of 461 bp (LsKDELR) and 566 bp (LsCOPB2) were synthesised using the MEGAscript RNAi Kit (Ambion), with purified PCR products including T7 promoters as templates.…”

Section: Rna Interferencementioning

confidence: 99%

“…To further investigate these effects on a molecular basis, the mRNA expression levels of the salmon louse egg proteins vitellogenin 1 (LsVit1), vitellogenin 2 (LsVit2) (Dalvin et al, 2011) and yolk associated protein (LsYAP) (Dalvin et al, 2009), as well as the digestive enzyme trypsin 1 (LsTryp1) (Kvamme et al, 2004a), were measured by qRT-PCR. As could be expected from the morphological analysis of the RNAi experiment, expression of yolk and yolk associated proteins; LsVit1, LsVit2 and LsYAP was disturbed in both LsKDELR and LsCOPB2 knock down lice (Fig.…”

Section: Downstream Effects Of Knock Down Of Lskdelr and Lscopb2mentioning

confidence: 99%

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RNA interference mediated knockdown of the KDEL receptor and COPB2 inhibits digestion and reproduction in the parasitic copepod Lepeophtheirus salmonis

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Nilsen

Dalvin

2014

Comparative Biochemistry and Physiology Part B: Biochemistry an

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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 knock down lice had a significantly higher mortality and failed to develop normally, while both LsCOPB2 and LsKDELR knock down 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.

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Section: Discussionmentioning

confidence: 72%

Section: Functional Analysis Of Lskdelr and Lscopb2mentioning

confidence: 95%

Section: Localisation Of Lskdelr and Lscopb2mentioning

confidence: 99%

Section: Rna Interferencementioning

confidence: 99%

Section: Downstream Effects Of Knock Down Of Lskdelr and Lscopb2mentioning

confidence: 99%

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RNA interference mediated knockdown of the KDEL receptor and COPB2 inhibits digestion and reproduction in the parasitic copepod Lepeophtheirus salmonis

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Nilsen

Dalvin

2014

Comparative Biochemistry and Physiology Part B: Biochemistry an

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A novel gene‐family involved in spermatophore generation in the economically important salmon louse Lepeophtheirus salmonis

Borchel

Nilsen

2018

Molecular Reproduction Devel

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The salmon louse (Lepeophtheirus salmonis) is an important parasite of Atlantic salmon (Salmo salar). It is widely spread in aquaculture facilities and leads to economic losses every year. As it has developed resistances against many common treatments, new control methods must be established. Here we characterize a novel gene family of the salmon louse, consisting of two genes, which has not been described in other species before. We analyzed temporal expression patterns of both genes, the localization of mRNA and protein. An RNAi mediated gene knockdown lead to information about the function of the protein. Overall, these two genes are expressed only in sperm ducts of male sea lice. The mucin-like proteins can additionally be found in the wall of spermatophores, which are responsible for sperm transfer to females. Knockdown showed that both genes are essential for successful fertilization of females. Overall, all results indicate that the two analyzed genes are necessary for reproduction in sea lice as they are essential for the formation of a wall surrounding the spermatophores, which is needed for fertilization. Therefore, we name them Mucin-like spermatophore wall protein 1 & 2 (MLSWP1 & MLSWP2). Analysis of sequence data from other copepod species suggests that MLSWPs are present in many copepod species and may also play a similar role in reproduction in those species.

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RNAi: An Instrument for Exploratory Experimentation

Solberg

2021

A Cognitive Ethnography of Knowledge and Material Culture

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Functional characterisation of the maternal yolk-associated protein (LsYAP) utilising systemic RNA interference in the salmon louse (Lepeophtheirus salmonis) (Crustacea: Copepoda)

Cited by 58 publications

References 25 publications

RNA interference mediated knockdown of the KDEL receptor and COPB2 inhibits digestion and reproduction in the parasitic copepod Lepeophtheirus salmonis

RNA interference mediated knockdown of the KDEL receptor and COPB2 inhibits digestion and reproduction in the parasitic copepod Lepeophtheirus salmonis

A novel gene‐family involved in spermatophore generation in the economically important salmon louse Lepeophtheirus salmonis

RNAi: An Instrument for Exploratory Experimentation

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