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Sequence analysis and identification of new isoform of EP4 receptors in different atlantic salmon tissues (Salmo salar L.) and its role in PGE2 induced immunomodulation in vitro.

Guo TC, Gamil AA, Koenig M, Evensen Ø - PLoS ONE (2015)

Bottom Line: In this study, we first sequenced EP4 receptor in different tissues and found that the presence of the 3nt deletion in the 5' untranslated region was accompanied by silent mutation at nt 668.While attempting to amplify the same sequence in TO cells (an Atlantic salmon macrophage-like cell line), we failed to obtain the full-length product.We further studied the immunomodulation effect of PGE2 in TO cells and found that PGE2 inhibited the induction of CXCL-10, CCL-4, IL-8 and IL-1β genes expression in a time dependent manner and without cAMP upregulation.

View Article: PubMed Central - PubMed

Affiliation: Norwegian University of Life Sciences, Faculty of Veterinary Medicine and Biosciences, Sea Lice Research Centre, P.O. Box 8146 Dep., 0033 Oslo, Norway.

ABSTRACT
PGE2 plays an important role in a broad spectrum of physiological and pathological processes mediated through a membrane-bound G protein-coupled receptor (GPCR) called EP receptor. In mammals, four subtypes of EP receptor (EP 1-4) are identified and each of them functions through different signal transduction pathways. Orthologous EP receptors have also been identified in other non-mammalian species, such as chicken and zebrafish. EP4 is the only identified PGE2 receptor to date in Atlantic salmon but its tissue distribution and function have not been studied in any detail. In this study, we first sequenced EP4 receptor in different tissues and found that the presence of the 3nt deletion in the 5' untranslated region was accompanied by silent mutation at nt 668. While attempting to amplify the same sequence in TO cells (an Atlantic salmon macrophage-like cell line), we failed to obtain the full-length product. Further investigation revealed different isoform of EP4 receptor in TO cells and we subsequently documented its presence in different Atlantic salmon tissues. These two isoforms of EP4 receptor share high homology in their first half of sequence but differ in the second half part with several deletion segments though the final length of coding sequence is the same for two isoforms. We further studied the immunomodulation effect of PGE2 in TO cells and found that PGE2 inhibited the induction of CXCL-10, CCL-4, IL-8 and IL-1β genes expression in a time dependent manner and without cAMP upregulation.

No MeSH data available.


Related in: MedlinePlus

cAMP induction post PGE2 treatment.Three cell lines, CHSE-214, RTG-2 and TO were used in this experiment. Cells were either treated with PGE2 (P) or left without treatment (C).
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pone.0120483.g008: cAMP induction post PGE2 treatment.Three cell lines, CHSE-214, RTG-2 and TO were used in this experiment. Cells were either treated with PGE2 (P) or left without treatment (C).

Mentions: To investigate the involvement of cAMP signaling pathway in PGE2 induced effect, the increase of cAMP concentration upon PGE2 treatment was assayed. In TO cells, no significant increase was detected. We included two additional fish-derived cell lines (CHSE-214 and RTG-2 cells, both from salmonid species) and both showed strong cAMP increase in response to PGE2 treatment (Fig. 8). We then included forskolin treatment as a positive control (including all said cells) and found that forskolin results in up-regulation of cAMP in CHSE-214 and RTG-2 while TO cells cAMP levels remained unchanged post stimulation (Fig. 9).


Sequence analysis and identification of new isoform of EP4 receptors in different atlantic salmon tissues (Salmo salar L.) and its role in PGE2 induced immunomodulation in vitro.

Guo TC, Gamil AA, Koenig M, Evensen Ø - PLoS ONE (2015)

cAMP induction post PGE2 treatment.Three cell lines, CHSE-214, RTG-2 and TO were used in this experiment. Cells were either treated with PGE2 (P) or left without treatment (C).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4383505&req=5

pone.0120483.g008: cAMP induction post PGE2 treatment.Three cell lines, CHSE-214, RTG-2 and TO were used in this experiment. Cells were either treated with PGE2 (P) or left without treatment (C).
Mentions: To investigate the involvement of cAMP signaling pathway in PGE2 induced effect, the increase of cAMP concentration upon PGE2 treatment was assayed. In TO cells, no significant increase was detected. We included two additional fish-derived cell lines (CHSE-214 and RTG-2 cells, both from salmonid species) and both showed strong cAMP increase in response to PGE2 treatment (Fig. 8). We then included forskolin treatment as a positive control (including all said cells) and found that forskolin results in up-regulation of cAMP in CHSE-214 and RTG-2 while TO cells cAMP levels remained unchanged post stimulation (Fig. 9).

Bottom Line: In this study, we first sequenced EP4 receptor in different tissues and found that the presence of the 3nt deletion in the 5' untranslated region was accompanied by silent mutation at nt 668.While attempting to amplify the same sequence in TO cells (an Atlantic salmon macrophage-like cell line), we failed to obtain the full-length product.We further studied the immunomodulation effect of PGE2 in TO cells and found that PGE2 inhibited the induction of CXCL-10, CCL-4, IL-8 and IL-1β genes expression in a time dependent manner and without cAMP upregulation.

View Article: PubMed Central - PubMed

Affiliation: Norwegian University of Life Sciences, Faculty of Veterinary Medicine and Biosciences, Sea Lice Research Centre, P.O. Box 8146 Dep., 0033 Oslo, Norway.

ABSTRACT
PGE2 plays an important role in a broad spectrum of physiological and pathological processes mediated through a membrane-bound G protein-coupled receptor (GPCR) called EP receptor. In mammals, four subtypes of EP receptor (EP 1-4) are identified and each of them functions through different signal transduction pathways. Orthologous EP receptors have also been identified in other non-mammalian species, such as chicken and zebrafish. EP4 is the only identified PGE2 receptor to date in Atlantic salmon but its tissue distribution and function have not been studied in any detail. In this study, we first sequenced EP4 receptor in different tissues and found that the presence of the 3nt deletion in the 5' untranslated region was accompanied by silent mutation at nt 668. While attempting to amplify the same sequence in TO cells (an Atlantic salmon macrophage-like cell line), we failed to obtain the full-length product. Further investigation revealed different isoform of EP4 receptor in TO cells and we subsequently documented its presence in different Atlantic salmon tissues. These two isoforms of EP4 receptor share high homology in their first half of sequence but differ in the second half part with several deletion segments though the final length of coding sequence is the same for two isoforms. We further studied the immunomodulation effect of PGE2 in TO cells and found that PGE2 inhibited the induction of CXCL-10, CCL-4, IL-8 and IL-1β genes expression in a time dependent manner and without cAMP upregulation.

No MeSH data available.


Related in: MedlinePlus