Central Institute of Freshwater Aquaculture, Kausalyaganga, Bhubaneswar 751002, India.
Fish Shellfish Immunol. 2011 Apr-May;30(4-5):1197-201. doi: 10.1016/j.fsi.2011.02.014. Epub 2011 Mar 6.
The innate immune response in fish represents an early and rapid defense against pathogens. The present study aims at looking into ontogeny of innate immune system in the teleost, Labeo rohita using RT-PCR based approach. Total RNA extracted from unfertilized and fertilized eggs, and hatchlings (hatched at 28 ± 2 °C) at 0, 1, 3, 6, 12, 24 h, and 3, 7, 16, 21, 31 days post-fertilization were subjected to RT-PCR using self-designed or earlier published primers to amplify some innate immune relevant genes (lysozyme C, lysozyme G, beta-2 microglobulin, toll-like receptor 22-like and transferrin). The constitutive expression of β-actin was detected in unfertilized eggs and further developmental stages. Transferrin and TLR22-like mRNA transcripts were detected by RT-PCR from 6 h post-fertilization to 31 day post-fertilization, whereas β-2 microglobulin transcripts were detected only from 7 day post-fertilization onwards. Lysozyme C mRNA transcripts were detected from 24 h post-fertilization to 31 day post-fertilization. Lysozyme G mRNA transcripts were detected early from unfertilized egg stage onwards. Similarly, tissues viz. intestine, heart, ovary, gill, spleen, muscle, liver, brain, skin, anterior kidney, posterior kidney, and blood collected from juveniles of rohu were subjected to detection of all above mentioned gene transcripts by RT-PCR. β2-microglobulin mRNA transcript was expressed in all tissues. Lysozyme C mRNA expression is confined to blood and posterior kidney only whereas lysozyme G mRNA is expressed in all tissues. TLR22-like mRNA is expressed in all tissues except ovary and liver whereas transferrin mRNA transcript is detected only in liver. Finally, all these information taken are likely to shed light on the ontogeny of innate immunity in L. rohita, which offers new insights to developmental biology when compared to higher vertebrates and also helpful in the development of preventive measures against problems concerning infectious diseases.
鱼类的先天免疫反应代表了对病原体的早期和快速防御。本研究旨在通过 RT-PCR 方法研究硬骨鱼罗非鱼（Labeo rohita）先天免疫系统的个体发生。从未受精和受精的卵以及在 28±2°C 孵化的幼体（孵化后 0、1、3、6、12、24 小时和 3、7、16、21、31 天）中提取总 RNA，并用自行设计或先前发表的引物进行 RT-PCR，以扩增一些先天免疫相关基因（溶菌酶 C、溶菌酶 G、β-2 微球蛋白、Toll 样受体 22 样和转铁蛋白）。β-肌动蛋白的组成型表达在未受精的卵和进一步的发育阶段中均被检测到。在受精后 6 小时至 31 天的时间内，通过 RT-PCR 检测到转铁蛋白和 TLR22 样 mRNA 转录本，而 β-2 微球蛋白转录本仅从受精后 7 天开始检测到。溶菌酶 C mRNA 转录本从受精后 24 小时到 31 天被检测到。溶菌酶 G mRNA 转录本从未受精的卵阶段开始就被早期检测到。同样，从罗非鱼幼体收集的组织（如肠、心脏、卵巢、鳃、脾脏、肌肉、肝脏、大脑、皮肤、前肾、后肾和血液）通过 RT-PCR 检测到所有上述基因转录本。β2-微球蛋白 mRNA 转录本在所有组织中均有表达。溶菌酶 C mRNA 的表达仅限于血液和后肾，而溶菌酶 G mRNA 则在所有组织中表达。TLR22 样 mRNA 在除卵巢和肝脏之外的所有组织中表达，而转铁蛋白 mRNA 转录本仅在肝脏中检测到。最后，所有这些信息都可能揭示罗非鱼先天免疫的个体发生，与高等脊椎动物相比，这为发育生物学提供了新的见解，并且对预防传染病相关问题也很有帮助。
