Biochemical characterization of three BLT receptors in zebrafish

Toshiaki Okuno, Tohru Ishitani, Takehiko Yokomizo

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

The leukotriene B4 (LTB4) receptor 1 (BLT1) is a high affinity receptor for LTB4, a chemotactic and inflammatory eicosanoid. The LTB4 receptor 2 (BLT2) was originally identified as a low affinity receptor for LTB4, and, more recently, as a high affinity receptor for 12-hydroxyheptadecatrienoic acid (12-HHT). The zebrafish BLT receptors have not been previously identified and the in vivo functions of these receptors have been unknown. In this paper, we describe one zebrafish BLT1-like receptor, Blt1, and two zebrafish BLT2-like receptors, Blt2a and Blt2b. Cells expressing Blt1 exhibited LTB4-induced intracellular [Ca2+] increases, inhibition of cAMP production, ligand-dependent [35S]GTPγS binding, and transforming growth factor-α (TGFα) shedding activity in a dose-dependent manner, similar to human BLT1. Cells expressing Blt2a and Blt2b exhibited 12-HHT- and LTB4-induced intracellular [Ca2+] increases, inhibition of cAMP production, [35S]GTPγS binding, and TGFa shedding activity, with a dose-dependency similar to human BLT2. Reverse transcription (RT)-PCR analysis and whole-mount in situ hybridization revealed that blt1, blt2a, blt2b, zebrafish LTA4 hydrolase (lta4h), and zebrafish 5-lipoxiganase (5lo) are expressed in zebrafish embryos. Knockdown of blt1 by morpholino antisense oligonucleotides resulted in delayed epiboly at gastrulation. Consistently, knockdown of lta4h, an enzyme mediating LTB4 production, induced a phenotype similar to knockdown of blt1. These results suggest that the LTB4-BLT1 axis is involved in epiboly in zebrafish development.

Original languageEnglish
Article numbere0117888
JournalPloS one
Volume10
Issue number3
DOIs
Publication statusPublished - Mar 4 2015

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Leukotriene B4 Receptors
Leukotriene B4
leukotrienes
Zebrafish
Danio rerio
receptors
Cells
Morpholinos
Eicosanoids
Antisense Oligonucleotides
Transforming Growth Factors
Transcription
hydrolases
Gastrulation
Ligands
calcium
transforming growth factors
Enzymes
eicosanoids
Reverse Transcription

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

Cite this

Biochemical characterization of three BLT receptors in zebrafish. / Okuno, Toshiaki; Ishitani, Tohru; Yokomizo, Takehiko.

In: PloS one, Vol. 10, No. 3, e0117888, 04.03.2015.

Research output: Contribution to journalArticle

Okuno, Toshiaki ; Ishitani, Tohru ; Yokomizo, Takehiko. / Biochemical characterization of three BLT receptors in zebrafish. In: PloS one. 2015 ; Vol. 10, No. 3.
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