Biochemical characterization of three BLT receptors in zebrafish.
Biochemical characterization of three BLT receptors in zebrafish.
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DOI：
10.1371/journal.pone.0117888
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Yokomizo T
中科院分区：
文献类型：
--
作者：
Okuno T;Ishitani T;Yokomizo T
白三烯B4（LTB 4）受体1（BLT 1）是LTB 4的高亲和力受体，LTB 4是一种趋化性和炎性类花生酸。LTB 4受体2（BLT 2）最初被鉴定为LTB 4的低亲和力受体，并且最近被鉴定为12-羟基十七碳三烯酸（12-HHT）的高亲和力受体。斑马鱼BLT受体以前尚未被鉴定，并且这些受体的体内功能一直是未知的。在本文中，我们描述了一个斑马鱼BLT 1样受体，Blt 1，和两个斑马鱼BLT 2样受体，Blt 2a和Blt 2b。表达Blt 1的细胞表现出LTB 4诱导的细胞内[Ca 2 +]增加、cAMP产生抑制、配体依赖性[35 S]GTPγS结合和转化生长因子-α（TGFα）脱落活性，这些均以剂量依赖性方式表现出，与人BLT 1相似。表达Blt 2a和Blt 2b的细胞表现出12-HHT和LTB 4诱导的细胞内[Ca 2 +]增加、cAMP产生抑制、[35 S]GTPγS结合和TGFα脱落活性，具有与人BLT 2相似的剂量依赖性。逆转录-PCR分析和整体原位杂交显示，blt 1，blt 2a，blt 2b，斑马鱼LTA 4水解酶（lta 4 h）和斑马鱼5-脂氧合酶（5lo）在斑马鱼胚胎中表达。通过吗啉代反义寡核苷酸敲低blt 1导致原肠胚形成延迟。一致地，敲低lta 4 h，一种介导LTB 4产生的酶，诱导了与敲低blt 1相似的表型。这些结果表明LTB 4-BLT 1轴参与了斑马鱼发育中的外皮。
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 TGFα 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.
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10.1084/jem.188.6.1063
发表时间：
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期刊：
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影响因子：
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2011-03-01
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发表时间：
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期刊：
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