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돼지 체외성숙난자에서 MMPs와 TIMPs의 발현 분석

Expression Analysis of Matrix Metalloproteinases and Tissue Inhibitor of Matrix Metalloproteinases from In Vitro Maturation Oocytes Complexes in Porcine

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한국동물번식학회 (The Korean Society of Animal Reproduction)
초록

Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, OC and COCs. Activity of MMP-2 in the OC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the COCs 36 hr. Expression of TIMP-3 protein in the COCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

목차
ABSTRACT   서론   재료 및 방법    난포란의 체취 및 성숙배양    Total RNA 분리 및 cDNA 합성    cDNA 정량 분석    Real-time PCR    Semiquantitative RT-PCR    Gelatin Zymography    Immunohistochemistry (IHC)    면역형광염색을 통한 MMP-2, MMP-9, TIMP-2, TIMP-3의 발현 양상 분석   결과    돼지 체외성숙란과 난구세포의 mRNA상의 MMPs, TIMPs의발현 양상    Zmorgraphy를 통한 MMP-2 및 MMP-9의 활성도 분석    면역염색방법을 이용한 MMP-2와 TIMP-2의 발현 위치분석    면역형광염색을 통해 MMPs와 TIMPs의 발현 양상 분석   고찰   인용문헌
저자
  • 김상환(한경대학교 생물환경정보통신전문대학원 동물생명공학 전공) | Sang-Hwan Kim
  • 강현아(한경대학교 생물환경정보통신전문대학원 동물생명공학 전공) | Hyun-Ah Kang
  • 김대승(한경대학교 생물환경정보통신전문대학원 동물생명공학 전공) | Dae-Seung Kim
  • 이명섭(메디키네틱스(주)) | Myeong-Seop Lee
  • 서강석(순천대학교 동물자원과학과) | Kang-Suk Seo
  • 민관식(한경대학교 생물환경정보통신전문대학원 동물생명공학 전공) | Kwan-Sik Min
  • 윤종택(한경대학교 생물환경정보통신전문대학원 동물생명공학 전공, 한경대학교 동물생명환경과학부 동물생명공학전공) | Jong-Taek Yoon Corresponding author