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长期饥饿和再投喂后凡纳滨对虾先天免疫和基因表达的调节

Modulation of innate immunity and gene expressions in white shrimp Litopenaeus vannamei following long-term starvation and re-feeding.

作者信息

Lin Yong-Chin, Chen Jiann-Chu, C Man Siti Nursafura, W Morni Wan Zabidii, N A Suhaili Awangku Shahrir, Cheng Sha-Yen, Hsu Chih-Hung

机构信息

Department of Aquaculture, College of Life Sciences, National Taiwan Ocean University, Keelung 202, Taiwan, ROC.

Department of Environmental Biology and Fisheries, College of Ocean Science and Resource, National Taiwan Ocean University, Keelung 202, Taiwan, ROC.

出版信息

Results Immunol. 2012 Jul 24;2:148-56. doi: 10.1016/j.rinim.2012.07.001. eCollection 2012.

Abstract

The survival rate, weight loss, immune parameters, resistance against Vibrio alginolyticus and white-spot syndrome virus (WSSV), and expressions of lipopolysaccharide- and ß-glucan-binding protein (LGBP), peroxinectin (PX), prophenoloxidase-activating enzyme (ppA), prophenoloxidase (proPO) I, proPO II, α2-macroglobulin (α2-M), integrin ß, heat shock protein 70 (HSP70), cytosolic manganese superoxide dismutase (cytMnSOD), mitochondrial manganese superoxide dismutase (mtMnSOD), and extracellular copper and zinc superoxide dismutase (ecCuZnSOD) were examined in the white shrimp Litopenaeus vannamei (8.18 ± 0.86 g body weight) which had been denied food (starved) for up to 14-28 days. Among shrimp which had been starved for 7, 14, 21, and 28 days, 100%, 90%, 71%, and 59% survived, and they lost 3.2%, 7.3%, 9.2%, and 10.4% of their body weight, respectively. Hyaline cells (HCs), granular cells (GCs, including semi-granular cells), the total haemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), and SOD activity significantly decreased in shrimp which had been starved for 1, 1, 1, 5, 14, and 3 days, respectively. The expression of integrin ß significantly decreased after 0.5-5 days of starvation, whereas the expressions of LGBP, PX, proPO I, proPO II, ppA, and α2-M increased after 0.5-1 days. Transcripts of all genes except ecCuZnSOD decreased to the lowest level after 5 days, and tended to background values after 7 and 14 days. Cumulative mortality rates of 7-day-starved shrimp challenged with V. alginolyticus and WSSV were significantly higher than those of challenged control-shrimp for 1-7 and 1-4 days, respectively. In another experiment, immune parameters of shrimp which had been starved for 7 and 14 days and then received normal feeding (at 5% of their body weight daily) were examined after 3, 6, and 12 h, and 1, 3, and 5 days. All immune parameters of 7-day-starved shrimp were able to return to their baseline values after 5 days of re-feeding except for GCs, whereas all parameters of 14-day-starved shrimp failed to return to the baseline values even with 5 days of re-feeding. It was concluded that shrimp starved for 14 days exhibited three stages of modulation of gene expression, together with reductions in immune parameters, and decreased resistance against pathogens.

摘要

对体重为(8.18±0.86克)的凡纳滨对虾进行长达14 - 28天的禁食处理,检测其存活率、体重减轻情况、免疫参数、对溶藻弧菌和白斑综合征病毒(WSSV)的抵抗力,以及脂多糖和β-葡聚糖结合蛋白(LGBP)、过氧化物酶(PX)、酚氧化酶原激活酶(ppA)、酚氧化酶原(proPO)I、proPO II、α2-巨球蛋白(α2-M)、整合素β、热休克蛋白70(HSP70)、胞质锰超氧化物歧化酶(cytMnSOD)、线粒体锰超氧化物歧化酶(mtMnSOD)和细胞外铜锌超氧化物歧化酶(ecCuZnSOD)的表达。在禁食7、14、21和28天的对虾中,存活率分别为100%、90%、71%和59%,体重分别减轻了3.2%、7.3%、9.2%和10.4%。在禁食1、1、1、5、14和3天的对虾中,透明细胞(HCs)、颗粒细胞(GCs,包括半颗粒细胞)、血细胞总数(THC)、酚氧化酶(PO)活性、呼吸爆发(RBs)和超氧化物歧化酶(SOD)活性显著下降。禁食0.5 - 5天后,整合素β的表达显著下降,而LGBP、PX、proPO I、proPO II、ppA和α2-M的表达在禁食0.5 - 1天后增加。除ecCuZnSOD外,所有基因的转录本在禁食5天后降至最低水平,并在禁食7天和14天后趋于背景值。用溶藻弧菌和WSSV攻击禁食7天的对虾,其累计死亡率在攻击后1 - 7天和1 - 4天分别显著高于攻击对照组对虾。在另一项实验中,对禁食7天和14天然后恢复正常投喂(每天按体重的5%投喂)的对虾,在投喂后3、6和12小时以及1、3和5天检测其免疫参数。禁食7天的对虾在重新投喂5天后,除GCs外,所有免疫参数均能恢复到基线值,而禁食14天的对虾即使重新投喂5天,所有参数也未能恢复到基线值。得出的结论是,禁食14天的对虾表现出基因表达调节的三个阶段,同时免疫参数降低,对病原体的抵抗力下降。

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