Fish macrophages express a cyclo-oxygenase-2 homologue after activation

J Zou, N F Neumann, J W Holland, M Belosevic, C Cunningham, C J Secombes, A F Rowley

Research output: Contribution to journalArticle

67 Citations (Scopus)

Abstract

In mammals, the increased generation of prostaglandins (PG) during the onset of inflammatory responses and activation of immune cell types has been attributed to the induction of a novel cyclo-oxygenase (COX) isoform, termed COX-2, which is distinct from the well-characterized constitutive activity (COX-1). Goldfish (Carassius auratus) macrophages exposed to bacterial lipopolysaccharide and leucocyte-derived macrophage-activating factor(s) showed a significant increase in the generation of the major COX product, PGE(2), within the first 6 h of stimulation. The selective COX-2 inhibitor, NS398, inhibited this elevated generation of PGE, whereas the basal level of this product synthesized by unstimulated macrophages was unaffected by such exposure. PGE generation by goldfish macrophages was similarly inhibited by the glucocorticoid, dexamethasone, and an inhibitor of protein synthesis, cycloheximide, suggesting that this stimulation may be due to an inducible enzyme equivalent to mammalian COX-2. The complete coding sequence of rainbow trout (Oncorhynchus mykiss) COX-2 was obtained by PCR. The gene contains a 61 bp 5'-untranslated region (UTR), a 1821 bp open reading frame and a 771 bp 3'UTR containing multiple copies of an mRNA instability motif (ATTTA). The predicted translation product had high homology to known mammalian and chicken COX-2 (83-84%) and COX-1 (77%) sequences. Reverse-transcriptase PCR with cDNA from control and bacterially challenged fish revealed that trout COX-2 expression was not constitutive but could be induced. Overall, these studies show for the first time that the inducible isoform of COX has a long evolutionary history, probably dating back to the evolution of fish over 500 million years ago.

Original languageEnglish
Pages (from-to)153-159
Number of pages7
JournalBiochemical Journal
Volume340
Publication statusPublished - 1999

Keywords

  • goldfish
  • leukotriene
  • Oncorhynchus mykiss
  • prostaglandins
  • trout
  • TROUT ONCORHYNCHUS-MYKISS
  • PROSTAGLANDIN ENDOPEROXIDE SYNTHASE-2
  • GOLDFISH KIDNEY LEUKOCYTES
  • RAINBOW-TROUT
  • HUMAN MONOCYTES
  • H SYNTHASE
  • CELL-LINE
  • CYCLOOXYGENASE-2
  • EICOSANOIDS
  • INDUCTION

Cite this

Zou, J., Neumann, N. F., Holland, J. W., Belosevic, M., Cunningham, C., Secombes, C. J., & Rowley, A. F. (1999). Fish macrophages express a cyclo-oxygenase-2 homologue after activation. Biochemical Journal, 340, 153-159.

Fish macrophages express a cyclo-oxygenase-2 homologue after activation. / Zou, J ; Neumann, N F ; Holland, J W ; Belosevic, M ; Cunningham, C ; Secombes, C J ; Rowley, A F .

In: Biochemical Journal, Vol. 340, 1999, p. 153-159.

Research output: Contribution to journalArticle

Zou, J, Neumann, NF, Holland, JW, Belosevic, M, Cunningham, C, Secombes, CJ & Rowley, AF 1999, 'Fish macrophages express a cyclo-oxygenase-2 homologue after activation', Biochemical Journal, vol. 340, pp. 153-159.
Zou, J ; Neumann, N F ; Holland, J W ; Belosevic, M ; Cunningham, C ; Secombes, C J ; Rowley, A F . / Fish macrophages express a cyclo-oxygenase-2 homologue after activation. In: Biochemical Journal. 1999 ; Vol. 340. pp. 153-159.
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AU - Zou, J

AU - Neumann, N F

AU - Holland, J W

AU - Belosevic, M

AU - Cunningham, C

AU - Secombes, C J

AU - Rowley, A F

PY - 1999

Y1 - 1999

N2 - In mammals, the increased generation of prostaglandins (PG) during the onset of inflammatory responses and activation of immune cell types has been attributed to the induction of a novel cyclo-oxygenase (COX) isoform, termed COX-2, which is distinct from the well-characterized constitutive activity (COX-1). Goldfish (Carassius auratus) macrophages exposed to bacterial lipopolysaccharide and leucocyte-derived macrophage-activating factor(s) showed a significant increase in the generation of the major COX product, PGE(2), within the first 6 h of stimulation. The selective COX-2 inhibitor, NS398, inhibited this elevated generation of PGE, whereas the basal level of this product synthesized by unstimulated macrophages was unaffected by such exposure. PGE generation by goldfish macrophages was similarly inhibited by the glucocorticoid, dexamethasone, and an inhibitor of protein synthesis, cycloheximide, suggesting that this stimulation may be due to an inducible enzyme equivalent to mammalian COX-2. The complete coding sequence of rainbow trout (Oncorhynchus mykiss) COX-2 was obtained by PCR. The gene contains a 61 bp 5'-untranslated region (UTR), a 1821 bp open reading frame and a 771 bp 3'UTR containing multiple copies of an mRNA instability motif (ATTTA). The predicted translation product had high homology to known mammalian and chicken COX-2 (83-84%) and COX-1 (77%) sequences. Reverse-transcriptase PCR with cDNA from control and bacterially challenged fish revealed that trout COX-2 expression was not constitutive but could be induced. Overall, these studies show for the first time that the inducible isoform of COX has a long evolutionary history, probably dating back to the evolution of fish over 500 million years ago.

AB - In mammals, the increased generation of prostaglandins (PG) during the onset of inflammatory responses and activation of immune cell types has been attributed to the induction of a novel cyclo-oxygenase (COX) isoform, termed COX-2, which is distinct from the well-characterized constitutive activity (COX-1). Goldfish (Carassius auratus) macrophages exposed to bacterial lipopolysaccharide and leucocyte-derived macrophage-activating factor(s) showed a significant increase in the generation of the major COX product, PGE(2), within the first 6 h of stimulation. The selective COX-2 inhibitor, NS398, inhibited this elevated generation of PGE, whereas the basal level of this product synthesized by unstimulated macrophages was unaffected by such exposure. PGE generation by goldfish macrophages was similarly inhibited by the glucocorticoid, dexamethasone, and an inhibitor of protein synthesis, cycloheximide, suggesting that this stimulation may be due to an inducible enzyme equivalent to mammalian COX-2. The complete coding sequence of rainbow trout (Oncorhynchus mykiss) COX-2 was obtained by PCR. The gene contains a 61 bp 5'-untranslated region (UTR), a 1821 bp open reading frame and a 771 bp 3'UTR containing multiple copies of an mRNA instability motif (ATTTA). The predicted translation product had high homology to known mammalian and chicken COX-2 (83-84%) and COX-1 (77%) sequences. Reverse-transcriptase PCR with cDNA from control and bacterially challenged fish revealed that trout COX-2 expression was not constitutive but could be induced. Overall, these studies show for the first time that the inducible isoform of COX has a long evolutionary history, probably dating back to the evolution of fish over 500 million years ago.

KW - goldfish

KW - leukotriene

KW - Oncorhynchus mykiss

KW - prostaglandins

KW - trout

KW - TROUT ONCORHYNCHUS-MYKISS

KW - PROSTAGLANDIN ENDOPEROXIDE SYNTHASE-2

KW - GOLDFISH KIDNEY LEUKOCYTES

KW - RAINBOW-TROUT

KW - HUMAN MONOCYTES

KW - H SYNTHASE

KW - CELL-LINE

KW - CYCLOOXYGENASE-2

KW - EICOSANOIDS

KW - INDUCTION

M3 - Article

VL - 340

SP - 153

EP - 159

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

ER -