Laboratory of Marine Immunobiology, Department of Animal Biology, University of Palermo, Via Archirafi, 18 Palermo, Italy.
Comp Biochem Physiol A Mol Integr Physiol. 2010 Sep;157(1):49-54. doi: 10.1016/j.cbpa.2010.05.003. Epub 2010 May 10.
Stressful conditions include a prompt release of corticosteroid hormones which can mediate gene expression through glucocorticoid receptors (GR). Since two seabass (Dicentrarchus labrax) GRs have been cloned and sequenced from peritoneal cavity cells (DlGR1) and liver (DlGR2), a comparative amino acid sequence analysis that included Haplochromis burtoni HbGRs, was carried out and homologies disclosed. The DlGR1 and DlGR2 deduced aminoacid sequences showed 61% identity (I) and 70% similarity (S). Moreover, DlGR2 was similar to HbGR2b (69% I, 73% S), and the DlGR1 to HbGR1 (72% I, 78% S). In addition, we examined the expression of the DlGRs after exogeneous cortisol inoculation into the peritoneal cavity, mimicking stress effects. At various times after the administration (3 h, 24 h, 1 week), gene expressions was evaluated in head kidney by real-time PCR. In addition, immunoblotting and densitometry analyses were performed with anti-DlGR1 antibodies. Although sea bass head kidney expressed both DlGR1 and DlGR2 they were differentially modulated by intraperitoneal implant of exogeneous cortisol.
应激条件包括糖皮质激素激素的迅速释放,这些激素可以通过糖皮质激素受体 (GR) 介导基因表达。由于已经从腹腔细胞 (DlGR1) 和肝脏 (DlGR2) 克隆和测序了两种鲈鱼 (Dicentrarchus labrax) 的 GR,因此进行了包括哈氏异唇鱼 HbGRs 的比较氨基酸序列分析,并揭示了同源性。DlGR1 和 DlGR2 推断的氨基酸序列显示出 61%的同一性 (I) 和 70%的相似性 (S)。此外,DlGR2 与 HbGR2b 相似 (69% I, 73% S),而 DlGR1 与 HbGR1 相似 (72% I, 78% S)。此外,我们在腹腔内接种外源性皮质醇以模拟应激效应后,检测了 DlGRs 的表达。在给药后不同时间 (3 h、24 h、1 周),通过实时 PCR 评估头肾中的基因表达。此外,还用抗 DlGR1 抗体进行了免疫印迹和密度测定分析。尽管鲈鱼头肾表达了 DlGR1 和 DlGR2,但它们被腹腔内植入的外源性皮质醇差异调节。