Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Cytological analysis of pregnancy-associated plasma protein-A expression in porcine neonatal testis

미성숙 돼지 정소에서 pregnancy-associated plasma protein-A의 발현의 세포학적 분석

  • Kim, Ji-youn (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Oh, Keon Bong (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Byun, Sung June (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Ock, Sun-A (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Lee, Hwi-Cheul (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Hwang, Seong-Su (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Park, SangHyun (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Ha, Wootae (Department of Stem Cell and Regenerative Biology, KIT, Konkuk University) ;
  • Woo, Jae-Seok (Animal Biotechnology Division, National Institute of Animal Science, RDA) ;
  • Song, Hyuk (Department of Stem Cell and Regenerative Biology, KIT, Konkuk University)
  • 김지윤 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 오건봉 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 변승준 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 옥선아 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 이휘철 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 황성수 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 박상현 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 하우태 (건국대학교) ;
  • 우제석 (농촌진흥청 국립축산과학원 동물바이오공학과) ;
  • 송혁 (건국대학교)
  • Received : 2018.09.04
  • Accepted : 2018.09.22
  • Published : 2018.09.30
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Abstract

The identification of biomarkers of a living tissues is essentially required to understand specific functions of the cells. In previous study, we reported IGFBP 3 as one of the putative biomarkers, by showing specific expression at porcine spermatogonial stem cells (SSCs) of early stage of porcine testis. In this study, we analyzed the expression of seven members of IGFBP family (IGFBPs) in SSCs and histological expression pattern of pregnancy-associated plasma protein-A (PAPP-A), which plays a role on the growth promoting enzyme by cleavage of IGFBPs in testis of 5 days old pig. RT-PCR analysis showed that IGFBP 1, 2, 3, 4, and 6 were expressed at high level specifically in porcine SSCs compared with whole testis. We performed immunohisotochemical staining of testis sections with PAPP-A and protein gene product 9.5 (PGP9.5) which are the known biomarkers for SSCs. We were not able to find co-expression of PAPP-A and PGP9.5; PAPP-A was expressed only in Sertoli cells and PGP9.5 expression was confirmed in spermatogonium. Additionally, we were able to confirm the GATA4 expression in Sertoli and Leydig cells as a regulator of Sertoli cell function was not detected PGP9.5 expressing cells, indicating indirect evidence of that cytolocalization of PAPP-A expression is limited in Sertoli cells. These results suggested that the PAPP-A expressed in Sertoli cells may play role on regulation of development and differentiation of testicular cells through the IGF axis in neonatal porcine testis.

생체 조직 내에서 표지인자의 발견은 해당 세포의 특성과 기능을 이해하는 데 매우 중요하다. 이전의 연구에서 본 연구진은 돼지정원줄기세포에서 특이적으로 IGFBP 3가 발현되어 표지인자로의 가능성을 보고한 바 있다. 본 연구에서는 IGFBP 3이외의 다른 family member가 정원줄기세포에서 특이적으로 발현하는 지와, 이의 발현을 조절하는 PAPP-A의 조직학적인 측면에서의 발현 양상을 5일령 돼지 정소에서 확인하였다. 그 결과 IGFBP 1, 2, 3, 4, 6의 발현은 정소 전체에서 발현되는 수준보다 돼지 정원줄기세포에서 더 높은 수준에서 발현하고 있음을 확인하였다. PAPP-A는 sertoli cell에서 특이적으로 발현하며, 정원줄기세포에서는 발현하지 않는 것을 PGP9.5와의 동시-조직염색으로 확인하였다. 이러한 결과는 Sertoli cell에서 발현하는 PAPP-A 단백질은 미성숙 돼지 정소에서 IGFBP family를 통해 정소 세포의 발달과 분화를 조절할 것으로 판단된다.

Keywords

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