Gene expression analysis of the innate immune system during early rearing and weaning of meagre (Argyrosomus regius)

Cindy Campoverde, Douglas J Milne, Christopher J Secombes, Alicia Estévez, Enric Gisbert, Karl B Andree*

*Corresponding author for this work

Research output: Contribution to journalArticle

Abstract

The present study is the first report of some representative innate immune genes in meagre (Argyrosomus regius) larvae. This study has specifically focused on the growth period from hatching to the juvenile stage, a critical time in marine fish development when reliance on innate immune mechanisms are required for survival. We report molecular cloning of partial open reading frames and expression patterns for some innate immune genes (c3, cox2, met, lyzc, mxp, myd88, nod2, nod3). In addition, phylogenetic analyses of some of the sequences obtained was performed where confusion among closely allied isoforms may have existed. These results show the met isoform from meagre is met II, an isoform more similar to a homolog described in Larimichthys crocea; lysozyme (lyzc) corresponds to the c-type and NOD isoforms (nod2, nod3) separate into different clades confirming their distinctness within a common evolutionary history. Gene expression profiles of innate genes were investigated, for nine developmental stages, from 8 days post-hatching (dph) to 120 dph. Present results demonstrated that c3, cox2, met II, lyzc, mxp, myd88, nod2, and nod3 were expressed in all stages of larval development and displayed distinct expression profiles in separate tissues (kidney, spleen gut and gill). Moreover, expression patterns suggested theses innate immune genes may be influenced by feeding practices, i.e. switching from live prey (rotifer and Artemia) and weaning onto an inert commercial diet. In addition to evaluating changes in gene expression during early development, this study evaluated the modulation of gene expression by means of in vivo trials in juveniles that were stimulated with PAMPs (LPS, poly I:C, β-glucan). These results revealed significant changes in mRNA levels of target genes in the kidney, spleen, gut and gills. However, expression profiles differed in magnitude depending on the stimulant and/or tissue. These results are discussed in terms of their relevance and potential application in aquaculture practices.

Original languageEnglish
Pages (from-to)819-832
Number of pages14
JournalFish & Shellfish Immunology
Volume94
Early online date6 Oct 2019
DOIs
Publication statusPublished - Nov 2019

Fingerprint

Argyrosomus regius
weaning
Immune system
immune system
Gene expression
rearing
gene expression
Genes
Protein Isoforms
gene
hatching
genes
gills
spleen
digestive system
kidneys
Larimichthys crocea
Tissue
fish development
polyinosinic-polycytidylic acid

Keywords

  • Innate immunity
  • Argyrosomus regius
  • Meagre
  • PAMP
  • Ontogeny
  • MOLECULAR CHARACTERIZATION
  • JAPANESE FLOUNDER
  • OXIDATIVE STRESS
  • LYSOZYME CDNA
  • SEA-BASS
  • COMPLEMENT-SYSTEM
  • C-TYPE
  • METALLOTHIONEIN GENE
  • FUNCTIONAL-ANALYSIS
  • ORANGE-SPOTTED GROUPER

Cite this

Gene expression analysis of the innate immune system during early rearing and weaning of meagre (Argyrosomus regius). / Campoverde, Cindy; Milne, Douglas J; Secombes, Christopher J; Estévez, Alicia; Gisbert, Enric; Andree, Karl B.

In: Fish & Shellfish Immunology, Vol. 94, 11.2019, p. 819-832.

Research output: Contribution to journalArticle

Campoverde, Cindy ; Milne, Douglas J ; Secombes, Christopher J ; Estévez, Alicia ; Gisbert, Enric ; Andree, Karl B. / Gene expression analysis of the innate immune system during early rearing and weaning of meagre (Argyrosomus regius). In: Fish & Shellfish Immunology. 2019 ; Vol. 94. pp. 819-832.
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T1 - Gene expression analysis of the innate immune system during early rearing and weaning of meagre (Argyrosomus regius)

AU - Campoverde, Cindy

AU - Milne, Douglas J

AU - Secombes, Christopher J

AU - Estévez, Alicia

AU - Gisbert, Enric

AU - Andree, Karl B

N1 - Acknowledgements This project has received funding from the European Union's Seventh Framework Programme for research, technological development and demonstration (KBBE-2013-07 single stage, GA 603121, DIVERSIFY). CC received funding from a Ph.D. grant awarded by the Secretaría Nacional de Educación Superior, Ciencia, Tecnología e Innovación (SENESCYT), Ecuador. We would like to thank G. Macià and M. Monllaó, S. Molas for their help in live prey production and meagre larval and juvenile rearing.

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N2 - The present study is the first report of some representative innate immune genes in meagre (Argyrosomus regius) larvae. This study has specifically focused on the growth period from hatching to the juvenile stage, a critical time in marine fish development when reliance on innate immune mechanisms are required for survival. We report molecular cloning of partial open reading frames and expression patterns for some innate immune genes (c3, cox2, met, lyzc, mxp, myd88, nod2, nod3). In addition, phylogenetic analyses of some of the sequences obtained was performed where confusion among closely allied isoforms may have existed. These results show the met isoform from meagre is met II, an isoform more similar to a homolog described in Larimichthys crocea; lysozyme (lyzc) corresponds to the c-type and NOD isoforms (nod2, nod3) separate into different clades confirming their distinctness within a common evolutionary history. Gene expression profiles of innate genes were investigated, for nine developmental stages, from 8 days post-hatching (dph) to 120 dph. Present results demonstrated that c3, cox2, met II, lyzc, mxp, myd88, nod2, and nod3 were expressed in all stages of larval development and displayed distinct expression profiles in separate tissues (kidney, spleen gut and gill). Moreover, expression patterns suggested theses innate immune genes may be influenced by feeding practices, i.e. switching from live prey (rotifer and Artemia) and weaning onto an inert commercial diet. In addition to evaluating changes in gene expression during early development, this study evaluated the modulation of gene expression by means of in vivo trials in juveniles that were stimulated with PAMPs (LPS, poly I:C, β-glucan). These results revealed significant changes in mRNA levels of target genes in the kidney, spleen, gut and gills. However, expression profiles differed in magnitude depending on the stimulant and/or tissue. These results are discussed in terms of their relevance and potential application in aquaculture practices.

AB - The present study is the first report of some representative innate immune genes in meagre (Argyrosomus regius) larvae. This study has specifically focused on the growth period from hatching to the juvenile stage, a critical time in marine fish development when reliance on innate immune mechanisms are required for survival. We report molecular cloning of partial open reading frames and expression patterns for some innate immune genes (c3, cox2, met, lyzc, mxp, myd88, nod2, nod3). In addition, phylogenetic analyses of some of the sequences obtained was performed where confusion among closely allied isoforms may have existed. These results show the met isoform from meagre is met II, an isoform more similar to a homolog described in Larimichthys crocea; lysozyme (lyzc) corresponds to the c-type and NOD isoforms (nod2, nod3) separate into different clades confirming their distinctness within a common evolutionary history. Gene expression profiles of innate genes were investigated, for nine developmental stages, from 8 days post-hatching (dph) to 120 dph. Present results demonstrated that c3, cox2, met II, lyzc, mxp, myd88, nod2, and nod3 were expressed in all stages of larval development and displayed distinct expression profiles in separate tissues (kidney, spleen gut and gill). Moreover, expression patterns suggested theses innate immune genes may be influenced by feeding practices, i.e. switching from live prey (rotifer and Artemia) and weaning onto an inert commercial diet. In addition to evaluating changes in gene expression during early development, this study evaluated the modulation of gene expression by means of in vivo trials in juveniles that were stimulated with PAMPs (LPS, poly I:C, β-glucan). These results revealed significant changes in mRNA levels of target genes in the kidney, spleen, gut and gills. However, expression profiles differed in magnitude depending on the stimulant and/or tissue. These results are discussed in terms of their relevance and potential application in aquaculture practices.

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KW - Argyrosomus regius

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KW - PAMP

KW - Ontogeny

KW - MOLECULAR CHARACTERIZATION

KW - JAPANESE FLOUNDER

KW - OXIDATIVE STRESS

KW - LYSOZYME CDNA

KW - SEA-BASS

KW - COMPLEMENT-SYSTEM

KW - C-TYPE

KW - METALLOTHIONEIN GENE

KW - FUNCTIONAL-ANALYSIS

KW - ORANGE-SPOTTED GROUPER

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DO - 10.1016/j.fsi.2019.10.009

M3 - Article

C2 - 31597086

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JO - Fish & Shellfish Immunology

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