Acerete L, Balasch J C, Castellana B, Redruello B, Roher N, Canario A V, Planas J V, MacKenzie S, Tort L
Department of Cell Biology, Physiology and Immunology, Universitat Autònoma de Barcelona, Facultat de Ciències, 08193 Bellaterra, Spain.
Comp Biochem Physiol B Biochem Mol Biol. 2007 Sep;148(1):32-43. doi: 10.1016/j.cbpb.2007.04.015. Epub 2007 Apr 30.
In order to determine the cortisol response after an immune challenge in the gilthead seabream (Sparus aurata), a cortisol receptor (GR) was cloned, sequenced and its expression determined after lipopolysaccharide (LPS) treatment. To clone the gilthead seabream GR (sbGR), consecutive PCR amplifications and screening of a pituitary cDNA library were performed. We obtained a clone of 4586 bp encoding a 784aa protein. Northern blot analysis from head kidney, heart and intestine revealed that the full length sbGR mRNA was approximately 6.5 Kb. A LPS treatment, used as an acute stress model, was employed to characterise the expression of sbGR and some selected genes involved in the immune response (IL-1beta, TNF-alpha, Mx protein, cathepsin D and PPAR-gamma). All genes were expressed in all tissues examined and responses were tissue and time dependent revealing differential gene expression profiles after LPS administration. Furthermore, analysis of plasma cortisol levels after LPS injection, showed an acute response to inflammatory stress with a significant increase two and six h after injection, recovering to basal levels 12 h post-stress in all LPS concentrations tested.
为了确定金头鲷(Sparus aurata)在免疫刺激后的皮质醇反应,克隆、测序了一种皮质醇受体(GR),并在脂多糖(LPS)处理后测定了其表达。为了克隆金头鲷GR(sbGR),进行了连续的PCR扩增和垂体cDNA文库筛选。我们获得了一个4586 bp的克隆,编码一种784个氨基酸的蛋白质。对头部肾脏、心脏和肠道进行的Northern印迹分析表明,全长sbGR mRNA约为6.5 Kb。使用LPS处理作为急性应激模型,以表征sbGR以及一些参与免疫反应的选定基因(IL-1β、TNF-α、Mx蛋白、组织蛋白酶D和PPAR-γ)的表达。所有基因均在所有检测组织中表达,反应具有组织和时间依赖性,显示出LPS给药后不同的基因表达谱。此外,LPS注射后血浆皮质醇水平分析显示,对炎症应激有急性反应,注射后2小时和6小时显著升高,在所有测试的LPS浓度下,应激后12小时恢复到基础水平。