Li Quanfu, Hu Lisi, Yang Ping, Zhang Qian, Waqas Yasir, Liu Tengfei, Zhang Linli, Wang Shuai, Chen Wei, Le Yuan, Ullah Shakeeb, Chen Qiusheng
Laboratory of Animal Cell Biology and Embryology College of Veterinary Medicine Nanjing Agricultural University Nanjing China.
Ecol Evol. 2015 Sep 23;5(19):4466-79. doi: 10.1002/ece3.1726. eCollection 2015 Oct.
The initiation of innate immunology system could play an important role in the aspect of protection for sperms long-term storage when the sperms got into oviduct of turtles and come into contact with epithelium. The exploration of TLR2/4 distribution and expression in oviduct during hibernation could help make the storage mechanism understandable. The objective of this study was to examine the gene and protein expression profiles in Chinese soft-shelled turtle during hibernation from November to April in the next year. The protein distribution of TLR2/4 was investigated in the magnum, isthmus, uterus, and vagina of the turtle oviduct using immunohistochemistry, and the gene expression of TLR2/4 was analyzed using quantitative real-time PCR (qRT-PCR). The results showed positive TLR2 protein expression primarily in the epithelium of the oviduct. TLR4 immunoreactivity was widely observed in almost every part of the oviduct, particularly in the epithelium and secretory gland membrane. Analysis of protein, mRNA expression revealed the decreased expression of TLR2/4 in the magnum compared with the isthmus, uterus, and vagina during hibernation. The protein and mRNA expression of TLR2 in the magnum, isthmus, uterus, and vagina was decreased in April compared with that in November. TLR4 protein and mRNA expression in the magnum, isthmus, uterus and vagina was decreased in November compared with that in April. These results indicated that TLR2/4 expression might protect the sperm from microbial infections. In contrast to the function of TLR2, which protects sperm during the early stages of hibernation, TLR4 might play a role in later stages of storage. The present study is the first to report the functions of TLR2/4 in reptiles.
当精子进入龟的输卵管并与上皮接触时，先天免疫系统的启动在精子长期储存的保护方面可能发挥重要作用。探索冬眠期间输卵管中TLR2/4的分布和表达有助于理解储存机制。本研究的目的是检测中华鳖在11月至次年4月冬眠期间的基因和蛋白质表达谱。采用免疫组织化学方法研究TLR2/4在龟输卵管的壶腹部、峡部、子宫和阴道中的蛋白质分布，并用实时定量PCR（qRT-PCR）分析TLR2/4的基因表达。结果显示，TLR2蛋白主要在输卵管上皮中呈阳性表达。TLR4免疫反应性在输卵管的几乎每个部位都广泛观察到，特别是在上皮和分泌腺膜中。蛋白质和mRNA表达分析显示，冬眠期间壶腹部的TLR2/4表达低于峡部、子宫和阴道。4月时，壶腹部、峡部、子宫和阴道中TLR2的蛋白质和mRNA表达低于11月。11月时，壶腹部、峡部、子宫和阴道中TLR4的蛋白质和mRNA表达低于4月。这些结果表明，TLR2/4的表达可能保护精子免受微生物感染。与TLR2在冬眠早期保护精子的功能不同，TLR4可能在储存后期发挥作用。本研究首次报道了TLR2/4在爬行动物中的功能。
