金定鸭不同组织和血液中TLR1、TLR2、TLR4、TLR5的表达差异分析

Analysis of TLR1,TLR2,TLR4 and TLR5 Expression in Different Tissues of Jinding Duck

试验旨在研究不同Toll样受体(Toll-like receptor,TLR)在鸭不同组织中的表达情况,选取300日龄雄性金定鸭10只,解剖后采集其血液及脾脏、肝脏、睾丸、肺脏、下丘脑、垂体、皮肤、腿肌、心脏、肾脏、胸肌、盲肠、小肠、胸腺,采用Primer Premier 5.0软件设计特异性引物,并用实时荧光定量PCR法检测TLR1、TLR2、TLR4、TLR5在不同组织中的相对表达情况。结果显示,各基因扩增产物的熔解曲线均有一特异性的单峰,无其他杂峰,说明引物的特异性较强,可以准确定量。4种目的基因与内参基因的扩增效率在101.4%~105.0%之间,均接近100%,相关系数(R2)为0.98~1.000。TLR1、TLR2、TLR4、TLR5 4种TLRs在金定鸭不同组织和血液中均有表达,但每种TLR受体在各组织中表达水平略有差异,其中TLR1在下丘脑中表达量最低,在胸肌中最高;TLR2在小肠中的表达量最低,在肺脏中最高;TLR4、TLR5在睾丸中的表达量最低,在皮肤中最高。以上结果说明,鸭TLRs在多种组织中能够广泛表达,本试验结果可为TLRs在鸭源感染过程中的作用机理研究提供科学依据。

The aim of this study was to explore the expression profiles of Toll-like receptor(TLR)in different tissues,and to lay the foundation of TLRs research for ducks.Ten male Jinding ducks of300days of age were slaughtered,and the blood sample and14different kinds of tissues(spleen,liver,testise,lung,hypothalamus,pituitary gland,skin,leg muscle,heart,kidney,chest muscle,caecum,small intestine and thymus)were collected.The Primer Premier5.0software was used to design specific primers and the quantitative Real-time PCR method was used to detect the relative expression levels of TLR1,TLR2,TLR4and TLR5mRNA.The results showed that there were only one specific peak on the melting curve of each gene amplification products,indicating that the specificity of the primers were strong.The amplification efficiency of the target genes was101.4%to105.0%and the correlation coefficient(R2)was0.98to1.000.The four kinds of TLRs expressed in all of the14tissues and blood with varying abundance in different tissues for each of the4kinds of TLRs.The expression levels of TLR1was lowest in hypothalamus,and highest in chest muscle.The expression levels of TLR2was lowest in small intestine,and highest in lung.The lowest expressed tissues for TLR4and TLR5were testis,and the highest expressed tissues were skin.The study indicated that TLRs were widely expressed in various tissues of duck,which laid the foundation for further study on the role mechanism of duck TLRs in the process of pathogen infection.