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미니돼지 정소에서 20α-HSD의 발현 및 특성화

Expression and Localization of 20α-Hydroxysteroid Dehydrogenase in Immature Pig Testis

  • 김정수 (한경대학교 미래융합기술대학원 동물생명공학전공, 동물생명환경과학과, 유전공학연구소) ;
  • 성훈기 (한경대학교 미래융합기술대학원 동물생명공학전공, 동물생명환경과학과, 유전공학연구소) ;
  • 밤바략차 뭉흐자야 (한경대학교 미래융합기술대학원 동물생명공학전공, 동물생명환경과학과, 유전공학연구소) ;
  • 심보웅 (국가영장류센타) ;
  • 허창기 (메디카이네틱스) ;
  • 강명화 (호서대학교 식품영양학과) ;
  • 민관식 (한경대학교 미래융합기술대학원 동물생명공학전공, 동물생명환경과학과, 유전공학연구소)
  • Kim, Jeong-Soo (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Resource Science, Institute of Genetic Engineering, Hankyong National University) ;
  • Seong, Hun-Ki (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Resource Science, Institute of Genetic Engineering, Hankyong National University) ;
  • Byambaragchaa, Munkhzaya (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Resource Science, Institute of Genetic Engineering, Hankyong National University) ;
  • Sim, Bo-Woong (National Primate Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Her, Chang-Gi (MediKinetics) ;
  • Kang, Myung-Hwa (Department of Food Science and Nutrition, Hoseo University) ;
  • Min, Kwan-Sik (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Resource Science, Institute of Genetic Engineering, Hankyong National University)
  • 투고 : 2017.02.24
  • 심사 : 2017.07.20
  • 발행 : 2017.07.30

초록

모든 포유동물에 있어서 임신유지와 준비를 위해서는 프로게스테론 호르몬은 필수적이다. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD)는 프로게스테론을 생물학적으로 불활성인 $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP)로 대사시키는 중요한 효소이며, 임신말기와 분만개시에 중요한 역할을 하는 효소이다. 본 연구에서는 메디카이네틱스 미니돼지의 정소에서 $20{\alpha}$-HSD의 발현에 대하여 분석하였다. 미니돼지 정소는 출생후 6, 9, 12, 18 및 21일에 채취하였다. RT-PCR에 의한 mRNA 분석결과 분만 후 6일의 정소에서도 검출되었으며, 21일의 정소에서 가장 높은 발현을 하는 것으로 검출되었다. 그러나, 분만 후의 태반에서는 검출되지 않았다. 웨스턴블릇에 의한 단백질 분석결과는 미니돼지 정소에서 특이적으로 37-kDa의 밴드가 검출되었다. 그러나 분만 후 6일의 정소에서는 단백질이 검출되지 않았다. 면역조직화학적 분석에 의하면 정소에서 $20{\alpha}$-HSD는 Sertoli 세포와 Leydig 세포에서 특이적으로 발현하였다. 따라서 이러한 연구결과는 분만 후 돼지 정소에서 $20{\alpha}$-HSD 단백질의 발현에 대한 연구로서는 처음으로 밝혔으며, 향후 돼지 정소에서의 $20{\alpha}$-HSD의 기능적인 역할에 대한 연구가 더욱 더 진행되어야 할 것으로 사료된다.

In all mammalian species, progesterone is essential in the preparation for and maintenance of pregnancy. $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone ($20{\alpha}$-OHP), and plays a crucial role in the termination of pregnancy and initiation of parturition. In this study, we characterized the expression and localization of $20{\alpha}$-HSDinthe testis of MediKinetics $Micropigs^{(R)}$. The testes were collected at days 6, 9, 12, 18, and 21 after birth. The $20{\alpha}$-HSD mRNA was found to be expressed in the testis at day 6 after birth by RT-PCR. The highest level of mRNA expression in the testis was detected on day 21 after birth. However, the mRNA was not detected in the placenta after parturition. Western blot for $20{\alpha}$-HSD reveal that the specific 37-kDa band was detected in immature pig testis. However, this band was not detected in testis tissue at day 6 after birth. In the immunohistochemical analysis of the testis, $20{\alpha}$-HSD was detected in the Sertoli cells and Leydig cells. Taken together, our study shows for the first time that the $20{\alpha}$-HSD mRNA and protein are expressed in pig testis after birth. Further investigation is required to elucidate the functional mechanisms of $20{\alpha}$-HSD in pig testis after birth.

키워드

참고문헌

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