热应激对性成熟猪睾丸TGF-β3和Claudin-11蛋白表达的影响

doi:10.11843/j.issn.0366-6964.2016.10.023

摘要/Abstract

摘要：

旨在探讨猪舍温度37～40 ℃热应激条件下，转化生长因子β3（TGF-β3）和Claudin-11在猪睾丸的表达与定位。6头性成熟长白公猪分成2组，3头为热应激组，置于温度控制在37～40 ℃的猪舍环境，每天3 h，连续7 d，每天于热处理结束后将猪赶回20～27 ℃正常猪舍环境中；另3头为对照组，饲养于20～27 ℃正常猪舍环境中。7 d后取睾丸组织，采用qRT-PCR、Western blotting及免疫组织化学对猪睾丸TGF-β3和Claudin-11的表达进行研究。qRT-PCR结果显示，与对照组相比，TGF-β3在热应激组的mRNA相对表达量显著升高（P<0.01），Claudin-11的mRNA相对表达量较对照组降低（P<0.05）。Western blotting结果显示，热应激处理组TGF-β3的蛋白表达较对照组升高（P＜0.05），Claudin-11的蛋白表达较对照组下降（P＜0.05）。免疫组织化学结果显示，热应激组TGF-β3免疫反应强阳性物定位于各级生精细胞和支持细胞，免疫阳性着色深度和范围高于对照组，提示热应激导致TGF-β3的表达增高；热应激组Claudin-11表达与对照组相比明显下降，对照组Claudin-11在血睾屏障位置呈明显的带状表达，而热应激组Claudin-11的表达局限在支持细胞周围，失去明显的血睾屏障带状表达。热应激影响猪睾丸TGF-β3和Claudin-11的表达与定位，提示热应激可能通过调节TGF-β3和Claudin-11的表达来影响精子发生。

Abstract:

The purposes of this study was to explore the expression and location of TGF-β3 and Claudin-11 in the boar testis under heat stress (37-40 ℃) and normal temperature (20-27 ℃).Six boars (Landrace，18 months of age) were used and divided into 2 groups.3 boars were homed in a thermo-controlled temperature (37-40 ℃，3 h daily，consecutive 7 d) house as a heat stress group.After heat treatment，the boars were driven back to normal temperature (20-27 ℃).The other 3 boars were homed in 20-27 ℃ house as a control group.7 days later，all boars were castrated and the testis tissues were harvested.qRT-PCR，Western blotting and immunohistochemistry were used to explore the changes of mRNA and protein in response to heat treatment.qRT-PCR showed that relative expression levels of TGF-β3 mRNA significantly increased （P<0.01），while relative expression levels of Claudin-11 mRNA decreased（P<0.05） in heat treatment group compared with the control.Western blotting found that the expression levels of TGF-β3 protein significantly increased（P<0.05）in heat treatment group，while the expression levels of Claudin-11 protein decreased（P<0.05）compared with the control.Immunohistochemistry results showed that：TGF-β3 immunoreactivity staining was observed in all stages of germ cells and Sertoli cells in both heat stress group and the control，and the depth and area of the positive staining in heat stress group were higher than that of the control；Claudin-11 immunoreactivity staining decreased in heat stress boars compared with the control in that Claudin-11 immunoreactivity staining localized in a consecutive strand area corresponding to the blood-testis barrier in the testis of control boars，while Claudin-11 immunoreactivity staining was limited to Sertoli cells and no obvious immunoreactivity strand that could be found in heat stress group.Then we could get a conclusion that heat stress damaged the sperm quality and spermatogenesis maybe partly via heat stress increased the expression of TGF-β3 and decreased the expression of Claudin-11 in boar testis.

中图分类号:

S852.1

张禛，范小瑞，席华明，梁亚俊，贺俊平. 热应激对性成熟猪睾丸TGF-β3和Claudin-11蛋白表达的影响[J]. 畜牧兽医学报, 2016, 47(10): 2136-2142.

ZHANG Zhen，FAN Xiao-rui，XI Hua-ming，LIANG Ya-jun，HE Jun-ping. Effect of Heat Stress on the Expression of TGF-β3 and Claudin-11 Protein in Mature Boar Testis[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(10): 2136-2142.

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