东北林蛙发育早期TLR2的表达及免疫应答特征

The Expression and Immune Response Characteristics of TLR2 at the Early Development Stage of Rana chensinensis

旨在研究东北林蛙发育早期TLR2的表达特征及其针对不同PAMPs的免疫应答特征。免疫组化染色及RT-PCR对11期至42期东北林蛙TLR2进行定位和定量检测;29期蝌蚪0.5μg/m L的LTA,LPS,Zymo A,Poly I∶C急性腹腔注射,RT-PCR检测24 h内TLR2 m RNA相对表达量变化。结果显示,TLR2在东北林蛙发育早期的组织中分布广泛,11期TLR2 m RNA相对表达量0.187±0.021,25期升至0.373±0.031后,42期降至0.170±0.020。24 h内未检测到LPS组和Zymo A组TLR2 m RNA相对表达量显著性变化(P>0.05),LTA组在4 h与12 h出现两次极显著性升高(P<0.01);Poly I∶C组于8 h至峰值(P<0.01)。东北林蛙发育早期TLR2的时空表达特征与其发育阶段的生境变化相适应,29期东北林蛙的TLR2对4种PAMPs识别和应答兼具了鱼类和哺乳动物的特征,体现了免疫系统在进化上的演变。

It was to study the expression and immune response characteristics of TLR2 at the early development stage of Rana chensinensis. Immunohistochemistry and RT-PCR were used to locate and quantify the TLR2 m RNA from stage 11 to stage 42 of Rana chensinensis. Tadpoles at stage 29 were injected with antigen LTA, LPS, Zymo A and Poly I∶C intra peritoneally. The expression changes of TLR2 m RNA were detected by RT-PCR within 24 h. TLR2 is widely distributed at the early development stage of Rana chensinensis. The relative expression of TLR2 m RNA increased form 0.187±0.021 to 0.373±0.031 at stage 25, and then returned to 0.170±0.020 progressively. The expression of TLR2 m RNA had no significant change in the LPS group and the Zymo A group（P0.05）. TLR2 m RNA of the LTE group peaked twice at 4 h 与 12 h after antigen injection（P0.01）;and the Poly I ：C group peaked at 8 h（P0.01）. The expression character of TLR2 matches with its living environment at the early development stage of Rana chensinensis.TLR2 did not responses to the challenge of LPS and Zymo A timely at the stage 29, on the other hand, TLR2 recognized and responsed to LTA and Poly I∶C group effectively. TLR2 of Rana chensinensis possesses the recognition and response characteristics to PAMPs both of fish and mammals.