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microPublication / Biology / Inducible expression of F48C1.9 encoding...
Inducible expression of F48C1.9 encoding a nematode specific secreted peptide in the adult epidermis upon Drechmeria fungal infection
Shizue Omi1 and Nathalie Pujol1
1Centre d'Immunologie de Marseille-Luminy (CIML), Aix-Marseille University, UM2, INSERM U1104, CNRS UMR7280, 13288 Marseille, France.
Correspondence to: Nathalie Pujol (pujol@ciml.univ-mrs.fr)
Figure 1: Inducible expression of F48C1.9 in the adult epidermis upon fungal infection. Young adult transgenic worms, carrying both F48C1.9p::GFP and col-12p::DsRed reporters were infected (infection) or not (control) for 18 hours at 25° C with the fungus Drechmeria coniospora, as described in (Pujol et al., 2008a). Representative images with a long pass GFP filter allowing simultaneous visualization of both red and green fluorescence of IG1514 frEx559 (A-B; bar, 10 µm.) and quantification of relative green fluorescence with the Copas Biosort of this strain (left) and a second independent strain IG1515 frEx560 (C; each point is a worm, bar indicates mean).

Description

Inducible immune responses are ubiquitous features of animal defences against infection. D. coniospora is a natural pathogen of nematode. It produces spore that attach to specific glycans on the worm’s cuticle surface coat (Rouger et al., 2014), pierce it and send hyphae throughout the organism. This triggers the rapid induction in the epidermis of genes from the nlp (for neuro-peptide-like protein) and cnc (caenacin) families. These genes encode structurally-related antimicrobial peptides (AMPs) (Pujol et al., 2012); their over-expression can lead to an increased resistance to infection are likely to be important in nature for the survival of worms (Taffoni and Pujol, 2015). Other genes are up-regulated after infection, including F48C1.9 encoding for a nematode specific small secreted peptide (Pujol et al., 2008b). To monitor its expression, we made transgenic strains containing the transcriptional reporter F48C1.9p::GFP with the co-injection marker col-12p::DsRed, which is constitutively expressed in the epidermis (Pujol et al., 2008a). Upon infection with D. coniospora, an induction of the GFP was observed in the epidermis of the worm (Figure 1). This work suggest that another uncharacterized peptide encoding gene is part of the immune response of the worm epidermis.

Reagents

Constructs: F48C1.9p::GFP was obtained by Gibson fusion of 1.1 kb of the F48C1.9 promoter in pPD95.75 amplified with primers CACAACGATGGATACGCTAAaatcaaattatgacgtgatgcc and GTTCTTCTCCTTTACTCATgtttgttgaagatctgatctg. Injected at 80 ng/µl together with col-12p::DsRed (Pujol et al, 2008a) at 20 ng/µl. Two independent transgenic strains were obtained IG1514 frEx559 and IG1515 frEx560[F48C1.9p::GFP; col-12p::DsRed.].

 

Acknowledgments

J Belougne for worm sorting, at the French National Functional Genomics platform, supported by the GIS IBiSA and Labex INFORM.

References

Pujol, N., Cypowyj, S., Ziegler, K., Millet, A., Astrain, A., Goncharov, A., Jin, Y., Chisholm, A.D., and Ewbank, J.J. (2008a). Distinct innate immune responses to infection and wounding in the C. elegans epidermis. Curr Biol 18, 481-489.
PubMed
Pujol, N., Davis, P.A., and Ewbank, J.J. (2012). The Origin and Function of Anti-Fungal Peptides in C. elegans: Open Questions. Front Immunol 3, 237.
PubMed
Pujol, N., Zugasti, O., Wong, D., Couillault, C., Kurz, C.L., Schulenburg, H., and Ewbank, J.J. (2008b). Anti-fungal innate immunity in C. elegans is enhanced by evolutionary diversification of antimicrobial peptides. PLoS Pathog 4, e1000105.
PubMed
Rouger, V., Bordet, G., Couillault, C., Monneret, S., Mailfert, S., Ewbank, J.J., Pujol, N., and Marguet, D. (2014). Independent Synchronized Control and Visualization of Interactions between Living Cells and Organisms. Biophysical journal 106, 2096-2104.
PubMed
Taffoni, C., and Pujol, N. (2015). Mechanisms of innate immunity in C. elegans epidermis. Tissue Barriers 3, e1078432.
PubMed

Funding

ANR-16-CE15-0001-01

Author Contributions

Shizue Omi: Investigation
Nathalie Pujol: Writing - original draft, Supervision.

Reviewed By

Anonymous

History

Received: January 24, 2019
Accepted: February 7, 2019
Published: February 8, 2019

Copyright

© 2019 by the authors. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Citation

Omi, S; Pujol, N (2019). Inducible expression of F48C1.9 encoding a nematode specific secreted peptide in the adult epidermis upon Drechmeria fungal infection. microPublication Biology. 10.17912/micropub.biology.000090.
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