The female-biased expression, transcriptional regulation and knock-down effect of insulin-like growth factor binding protein 7 in Chinese tongue sole, Cynoglossus semilaevis
The female-biased expression, transcriptional regulation and knock-down effect of insulin-like growth factor binding protein 7 in Chinese tongue sole, Cynoglossus semilaevis
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DOI：
10.1016/j.aquaculture.2022.737956
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发表时间：
2022-01
期刊：
影响因子：
4.5
通讯作者：
Qiang Yang;Xihong Li;Rui Shi;Peng Cheng;Na Wang;Songlin Chen
中科院分区：
文献类型：
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作者：
Qiang Yang;Xihong Li;Rui Shi;Peng Cheng;Na Wang;Songlin Chen
胰岛素样生长因子结合蛋白7(IGFBP7)是胰岛素样生长因子结合蛋白(IGFBP)超家族成员之一，参与胰岛素样生长因子(IGF)的调节和胰岛素信号转导。最近发现igfbp7在半滑舌鲨(Cynoglossus Semilaevis)的性腺中存在雌性偏向的表达，这是一种表现出雌性偏向的性二型性的海洋比目鱼。本研究旨在探讨igfbp7在FC生长和性别分化中的作用。首先，克隆了半乳杆菌igfbp7基因，编码序列为666hp，编码221个氨基酸。系统发育和结构分析表明，c.Semilaevis igfbp7与大西洋大比目鱼(Hippoglossusherglossus)、帆鳍鱼(Poecilia Laitpinna)等聚为一类。随后的时间和空间表达分析表明，fbp7主要在雌性性腺表达。在不同的性腺发育阶段，igfbp7的表达在6月龄达到最高水平。甲基组数据显示，在性腺的整个基因区域，女性fbp7的甲基化水平低于男性和假瘤。双荧光素酶实验表明，c.Semilaevis igfbp7启动子具有较强的转录活性。此外，垂体特异性正转录因子1(POU1F1)和类固醇激素-睾酮显著增强了IGFBP7启动子的转录活性。体外转染实验显示，IGFBP7在核因子-κB信号通路上被激活。最后观察胰岛素样生长因子1受体(IGF1R)、丝氨酸/苏氨酸激酶1(AKT)和热休克60蛋白1(Hspd1)的激活。将小干扰RNA(SiRNA)导入半卵巢细胞。这些结果为fbp7通过影响IGFR、AKT和NF-κB信号在生长调节中的作用提供了因果线索。
Insulin-like growth factor binding protein 7 (igfbp7) is a member of the insulin-like growth factor binding protein (IGFBP) superfamily, which is involved in the regulation of insulin-like growth factor (IGF) and insulin signal transduction. Recently,igfbp7was found to be female-biased expressed in the gonad of Chinese tongue sole (Cynoglossus semilaevis), a marine flatfish exhibited female-biased sexual dimorphism. This study aimed to explore the roles ofigfbp7in the growth and sex differentiation ofC. semilaevis.Firstly,C. semilaevis igfbp7was cloned, and the coding sequence (CDS) was 666 bp encoding 221 amino acids. The phylogenetic and structural analyses revealed thatC. semilaevis igfbp7was clustered with Atlantic halibut (Hippoglossushippoglossus), sailfin molly (Poecilia latipinna),and so on. The subsequent temporal and spatial expression analyses showed thatigfbp7was mainly expressed in female gonad. During the different gonad developmental stages, the expression ofigfbp7displayed the highest level at 6-month. Methylome data showed that femaleigfbp7exhibited lower methylation levels than male and pseudomale across the whole gene region in the gonad. Dual-luciferase experiment revealed thatC. semilaevis igfbp7promoter exhibited strong transcription activity. In addition, pituitary-specific positive transcription factor 1 (POU1F1) and steroid hormone-testosterone significantly enhanced the transcriptional activity of the igfbp7 promoter.In vitrotransfection experiment revealed the activation ofigfbp7on the nuclear factor kappa B (NF-κB) signaling pathway. Finally, the activation of insulin-like growth factor 1 receptor (igf1r), serine/threonine kinase 1 (akt) and heat shock 60 protein 1 (hspd1) was observed inC. semilaevisovary cells transfected byigfbp7small interfering RNA (siRNA). These results provided causal clues forigfbp7in growth regulation by influencingigf1r,aktand NF-κB signal.