短期(1、6和24小时)空气暴露应激(1分钟)的应激和免疫相关效应,用anguillarum弧菌疫苗接种,两种应激源组合在Sparusaurata的肝脏和脾脏中进行评估,Daniorerio和Onchorhynchusmykiss.通过RT-qPCR分析了免疫(白细胞介素1β:il1β;肿瘤坏死因子α:tnfα;白细胞介素10:il10;肿瘤生长因子β:tgfβ1;免疫球蛋白M:igm;溶菌酶:lys;补体蛋白c3:c3)和应激相关基因(糖皮质激素受体:gr;热休克蛋白70:hsp70和烯醇化酶)的表达谱。通过放射免疫测定评估皮质醇水平。发现肝脏和脾脏中的基因表达模式在物种之间以时间和器官依赖性方式差异调节。在Seabream,记录到更高的IL1β驱动的炎症反应.在斑马鱼中,空气暴露压力而不是单独的浴疫苗调节肝脏和脾脏免疫转录的大部分变化。强调和接种的鳟鱼显示出基因表达的中间模式,肝脏中免疫相关基因的上调较低,并且脾脏中hsp70和烯醇化酶的表达没有变化(如在海马中观察到的,而在斑马鱼中没有观察到)。在空气暴露之后,血浆皮质醇水平在应激后1小时升高(hps),然后在O.mykiss和D.rerio中在6hps时降低。相比之下,在S.aurata中,皮质醇水平在6hps时仍然较高,表明对这种应激源的反应程度更高。当鱼暴露于空气暴露加沐浴疫苗的组合时,O.mykiss和S.aurata的皮质醇水平也增加了1和6hps,并在24hps时恢复到基础水平,而在D.rerio中,对两种应激源的组合反应较高。此外,V.anguillarum细菌疫苗接种仅在D.rerio触发皮质醇分泌,提示下丘脑-垂体-肾间轴的反应性较高。总的来说,比较组织转录反应性,与脾脏相比,发现肝脏对处理压力的反应更有牵连。这些结果还表明,物种特异性反应解释了这些鱼类的肝脏和脾脏中应激和免疫发作的偏差。
The stress and immune-related effects of short-term (1, 6 and 24 h) air exposure stress (1 min), bath vaccination with Vibrio anguillarum bacterin, and both stressors combined were evaluated in liver and spleen of Sparus aurata, Danio rerio and Onchorhynchus mykiss. Expression profiles of immune (interleukin 1 beta: il1β; tumor necrosis factor alpha: tnfα; interleukin 10: il10; tumor growth factor beta: tgfβ1; immunoglobulin M: igm; lysozyme: lys; complement protein c3: c3) and stress-related genes (glucocorticoid receptor: gr; heat shock protein 70: hsp70; and enolase) were analysed by RT-qPCR. Cortisol level was assessed by radioimmunoassay. The gene expression patterns in liver and spleen were found to be differentially regulated in a time- and organ-dependent manner among species. In seabream, a higher il1β-driven inflammatory response was recorded. In zebrafish, air exposure stress but not bath vaccination alone modulated most of the changes in liver and spleen immune transcripts. Stressed and vaccinated trout showed an intermediate pattern of gene expression, with a lower upregulation of immune-related genes in liver and the absence of changes in the expression of hsp70 and enolase in spleen (as it was observed in seabream but not in zebrafish). Following air exposure, cortisol levels increased in plasma 1 h post-stress (hps) and then decreased at 6 hps in O. mykiss and D. rerio. By contrast, in S.aurata the cortisol level remained higher at 6 hps suggesting a greater degree of responsiveness to this stressor. When fish were exposed to combined air exposure plus bath vaccination cortisol levels were also augmented at 1 and 6 hps in O. mykiss and S.aurata and restored to basal level at 24 hps, whereas in D. rerio the response was higher in response to the combination of both stressors. In addition, V. anguillarum bacterin vaccination triggered cortisol secretion only in D. rerio, suggesting a greater responsiveness of D. rerio hypothalamic-pituitary-interrenal axis. Overall, comparing the tissue transcription responsiveness, liver was found to be more implicated in the response to handling stress compared to spleen. These results also indicate that a species-specific response accounts for the deviations of stress and immune onset in the liver and spleen in these fish species.