Scientia Agricultura Sinica ›› 2017, Vol. 50 ›› Issue (5): 924-931.doi: 10.3864/j.issn.0578-1752.2017.05.015

• ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT • Previous Articles     Next Articles

Effect of Heat Stress on the Expression of Cyt-C and Caspase-3 in Boar Testis

FAN XiaoRui, ZHANG Zhen, XI HuaMing, LIANG YaJun, HE JunPing   

  1. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, Shanxi
  • Received:2016-05-11 Online:2017-03-01 Published:2017-03-01

Abstract: 【Objective】 Elevated ambient temperature has detrimental effects on sperm quality in mammalian species including pig. But the molecular mechanism has not yet well understood. The aims of present study were to investigate the effect of heat stress on the expression of Cyt-C and Caspase-3 in boar testis. 【Method】 Nine mature boars (Landrace, 18 months of age) were used in the study. Three boars were subjected to 37-40℃ for 3 h daily for 7 days (heat stress 7 d, HS7d). Three boars were subjected to 37-40℃ for 3 h daily for 42 days (heat stress 42 d, HS42d). Following daily heat treatment the boars were driven back to normal house (20-27℃). The other 3 boars were kept in the normal house as control group (20-27℃). All boars were castrated and the testes were harvested. Western blot, QRT-PCR, and immunohistochemistry were used to analyze the expression of Cyt-C and Caspase-3. 【Result】 QRT-PCR showed that mRNA expression of Cyt-C and Caspase-3 were all increased in HS7d group and HS42d group compared with the control. And the relative expression of Cyt-C and Caspase-3mRNA in HS7d group was the highest. Western blot showed that the protein expression of Cyt-C and Caspase-3 was all increased in HS7d and HS42d as well, compared with the control group. And the protein expression of Cyt-C and Caspase-3 in HS7d group was the highest. Immunohistochemistry found that Cyt-C localized in the cytoplasm of Leydig cells, Sertoli cells and all stages of germ cells. In spermatogonia, the Cyt-C expression was low, while in postmeiosis germ cells, the expression of Cyt-C was higher. Heat treatment resulted in a more diffuse cytoplasmic localization of Cyt-C, which suggested that Cyt-C was released from mitochondria to the cytoplasm. In Leydig cells and spermatogonia, the Caspase-3 expression was low. A large number of postmeiosis germ cells showed Caspase-3 positive immunoreactive staining. Heat stress resulted in some Sertoli cells Caspase-3 positive staining and Caspase-3 positive with nucleus staining increased.【Conclusion】Elevated ambient temperature induced the increase of the expression of Cyt-C and Caspase-3, and the changes of cellular location, which indicated that the expression change of Cyt-C and Caspase-3 may be associated with the sperm quality decrease induced by heat stress.

Key words: heat stress, boar, testis, Cyt-C, Caspase-3

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