School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, People's Republic of China.
Fish Shellfish Immunol. 2012 Aug;33(2):418-26. doi: 10.1016/j.fsi.2012.05.033. Epub 2012 Jun 7.
In mammals, retinoid-related orphan receptors (ROR) consist of three members as RORα, RORβ and RORγ. It is well known that RORα plays a critical role in cerebellum development while RORγt directs T helper 17 (Th17) cell differentiation. So far, the knowledge on fish ROR family is limited as only zebrafish ROR family members have been characterized, showing that they play roles in embryonic and cerebellar development. In this study, we have cloned two paralogues for RORα (RORα1 and RORα2) and RORγ (RORγ1 and RORγ2) from grass carp (Ctenopharyngodon idellus). Phylogenetic analysis showed that grass carp RORα and RORγ were grouped in the clade of zebrafish RORα and RORγ, respectively. Real-time RT-PCR assay revealed that these four ROR transcripts exhibited similar expression patterns, in particular the high levels in pituitary, brain and some immune-related tissues, suggesting that all of them may play a role in endocrine and immune system of teleost. To explore the immune roles of grass carp RORα and RORγ, their expression was detected in periphery blood lymphocytes (PBLs) challenged by immune stimuli. Results showed that both RORα and RORγ mRNA levels were up-regulated by PHA but not LPS in PBLs, suggesting that their expression may be subject to different immune processes. In the same cell model, poly I:C stimulation induced RORγ1/2 but not RORα1/2 expression, pointing to the different roles of grass carp RORα and RORγ in immune response. Consistently, bacterial challenge significantly up-regulated the expression of these four ROR genes in spleen, headkidney and thymus. These results not only contribute to elucidate the roles of ROR in fish immunity but also facilitate to further clarify the existence of Th17-like cells in fish.
在哺乳动物中，维甲酸相关孤儿受体（ROR）由三个成员组成，分别是 RORα、RORβ 和 RORγ。众所周知，RORα 在小脑发育中起着关键作用，而 RORγt 则指导辅助性 T 细胞 17（Th17）细胞分化。到目前为止，鱼类 ROR 家族的知识还很有限，因为只有斑马鱼 ROR 家族成员被鉴定出来，表明它们在胚胎和小脑发育中发挥作用。在这项研究中，我们从草鱼（Ctenopharyngodon idellus）中克隆了两个 RORα（RORα1 和 RORα2）和 RORγ（RORγ1 和 RORγ2）的同源物。系统进化分析表明，草鱼 RORα 和 RORγ 分别被归类为斑马鱼 RORα 和 RORγ 的分支。实时 RT-PCR 检测显示，这四个 ROR 转录本具有相似的表达模式，特别是在垂体、大脑和一些免疫相关组织中表达水平较高，表明它们可能都在硬骨鱼的内分泌和免疫系统中发挥作用。为了探讨草鱼 RORα 和 RORγ 的免疫作用，我们检测了它们在外周血淋巴细胞（PBLs）受到免疫刺激后的表达情况。结果表明，PHA 而非 LPS 可诱导 PBLs 中 RORα 和 RORγ 的 mRNA 水平上调，表明它们的表达可能受到不同的免疫过程调控。在相同的细胞模型中，poly I:C 刺激诱导了 RORγ1/2 的表达，但没有诱导 RORα1/2 的表达，这表明草鱼 RORα 和 RORγ 在免疫反应中发挥了不同的作用。同样，细菌刺激显著上调了脾脏、头肾和胸腺中这四个 ROR 基因的表达。这些结果不仅有助于阐明 ROR 在鱼类免疫中的作用，还有助于进一步阐明鱼类中 Th17 样细胞的存在。
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