Heinecke Rasmus D, Chettri Jiwan K, Buchmann Kurt
Laboratory of Aquatic Pathobiology, Department of Veterinary Disease Biology, Faculty of Health and Medical Sciences, University of Copenhagen, Stigbøjlen 7, DK-1870 Frederiksberg C, Denmark.
Fish Shellfish Immunol. 2014 May;38(1):25-33. doi: 10.1016/j.fsi.2014.02.010. Epub 2014 Feb 21.
The ontogenetic development of the immune system was studied during the egg phase and the early post-hatch period of rainbow trout. Quantitative real-time PCR (qPCR) was used to assess the timing and degree of expression of 9 important immune relevant genes and EF1-α. Further, immunohistochemical staining using monoclonal antibodies was applied on rainbow trout embryos and larvae in order to localize five different protein molecules (MHCII, CD8, IgM, IgT and SAA) in the developing tissue and immune organs. Maternally transferred transcripts of EF1-α mRNA were detected in the unfertilized egg. Early onset of expression was seen for all immune genes at very low levels. The amount of mRNA slowly increased and peaked around and after hatching. The highest increases were seen for MHCII, C3, C5 and SAA. Immunohistochemistry using five monoclonal antibodies showed positive staining from day 84 post fertilization. Skin, gills, intestine, pseudobranch and thymus showed reactivity for MHCII, thymus for CD8, gill mucus for IgT and pseudobranch and cartilage associated tissue for SAA. The importance of detected factors for early protection of eggs and larvae is discussed.
在虹鳟鱼的卵期和孵化后早期，对其免疫系统的个体发育进行了研究。采用定量实时PCR（qPCR）来评估9个重要免疫相关基因和EF1-α的表达时间和表达程度。此外，使用单克隆抗体进行免疫组织化学染色，对虹鳟鱼胚胎和幼体进行检测，以便在发育中的组织和免疫器官中定位5种不同的蛋白质分子（MHCII、CD8、IgM、IgT和SAA）。在未受精卵中检测到母源转移的EF1-α mRNA转录本。所有免疫基因在极低水平时就出现了早期表达。mRNA的量缓慢增加，并在孵化前后达到峰值。MHCII、C3、C5和SAA的增加最为显著。使用5种单克隆抗体进行的免疫组织化学显示，受精后第84天开始出现阳性染色。皮肤、鳃、肠道、伪鳃和胸腺对MHCII呈阳性反应，胸腺对CD8呈阳性反应，鳃黏液对IgT呈阳性反应，伪鳃和软骨相关组织对SAA呈阳性反应。文中讨论了所检测因素对卵和幼体早期保护的重要性。
