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