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Positive selection pressure within teleost toll-like receptors tlr21 and tlr22 subfamilies and their response to temperature stress and microbial components in zebrafish

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Abstract

Toll-like receptors (TLRs) play a crucial role in host defence, since they trigger immune response following recognition of pathogen-associated molecular patterns (PAMPs) in potential infectious agents. TLRs have been found in numerous organisms, including mammals, birds and teleosts. Some TLR members are commonly retained across all species, whilst others were lost, gained or diverged independently during evolution. Our knowledge about the evolution and specific functions of tlr21, tlr22 and tlr23 in teleosts are still scarce. Phylogenetic analysis of 18 tlr13, tlr21, tlr22 and tlr23 genes from 9 different fish species divided them in two groups. All tlr21 genes were under the first clade, while the second comprised tlr22, tlr23 and tlr13 from Atlantic salmon. Evidence of positive selection was detected at three sites within the leucine-rich repeat regions of Tlr22, which may influence PAMP recognition. Immunostimulation experiments revealed that expression of zebrafish tlr22 is modulated by several unrelated PAMPs. Up to a 3-fold increase in tlr21 and tlr22 expression was detected in larvae exposed to immunostimulants such as lipopolysaccharide, peptidoglycan or poly I:C. We found that zebrafish tlrs are expressed mainly in immune-related organs, such as spleen and kidney as well as in testis and temperature stress did not have an effect on the expression of tlr21 and tlr22 in the early stages of development in zebrafish larvae. Our data indicates that these teleost tlrs may play a role in innate host defence. In particular, tlr22 is evolving under positive selection, which indicates functional diversification and adaptation of the response to different PAMPs.

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Abbreviations

Tlr:

Toll-like receptor

PAMPs:

Pathogen-associated molecular patterns

LRR:

Leucine-rich repeat

LPS:

Lipopolysaccharide from Salmonella enterica typhimurium

IC:

Polyinosinic-polycytidylic acid potassium salt

PG:

Peptidoglycan from Staphylococcus aureus

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Acknowledgments

Mr. Sundaram’s PhD is funded by a scholarship from the University of Nordland (Norway). We are grateful to Dr. Alessia Giannetto, Dr. Kazue Nagasawa, Ms. Catarina Campos and Mr. Spyros Kollias for their help in laboratory work and fish husbandry.

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Authors and Affiliations

  1. Faculty of Biosciences and Aquaculture, University of Nordland, 8049, Bodø, Norway

    Arvind Y. M. Sundaram, Viswanath Kiron & Jorge M. O. Fernandes

  2. Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Ceredigion, SY23 3DA, UK

    Sonia Consuegra

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  1. Arvind Y. M. Sundaram
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  2. Sonia Consuegra
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  4. Jorge M. O. Fernandes
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Correspondence to Jorge M. O. Fernandes.

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Sundaram, A.Y.M., Consuegra, S., Kiron, V. et al. Positive selection pressure within teleost toll-like receptors tlr21 and tlr22 subfamilies and their response to temperature stress and microbial components in zebrafish. Mol Biol Rep 39, 8965–8975 (2012). https://doi.org/10.1007/s11033-012-1765-y

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