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Korean Journal of Reproductive Medicine 2009;36(4):265-274.
Published online December 1, 2009.
Profiles of microRNAs in Mice Follicles According to Gonadotropins during in vitro Culture.
Yong Jin Kim, Seung Yup Ku, Yoon Young Kim, Sun Kyung Oh, Seok Hyun Kim, Young Min Choi, Shin Yong Moon
1Department of Obstetrics and Gynecology, College of Medicine, Seoul National University, Korea.
2Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University, Korea.
Abstract
OBJECTIVE
MicroRNAs (miR) are known to repress target genes at post-transcriptional level and play important roles in development and maturation of cell. However, the expression profiles of miR during ovarian follicle maturation have not been fully elucidated. Here, we designed this study to investigate the expression profiles of miR in oocytes and granulose cells (G-cells) after in vitro culture according to gonadotropins and adding hCG. METHODS: Ovaries from 12-day-old mice (C57BL6) were removed and preantral follicles were isolated and cultured in 20 micronL-drop of culture media with supplementation of either rFSH, rLH, or rFSH + rLH. After their full maturation, follicles were incubated with rhCG and rEGF. RNA was isolated from oocytes and G-cells, and real-time PCR were performed with primers of miR known to be expressed in the mouse ovary (mmu-miR-16, -miR-27a, -miR-126, -miR-721). RESULTS: FSH + LH group showed the highest ovulation and MII rates among gonadotropin groups. The profiles of miRs in oocytes and G-cells differed according to gonadotropin groups and adding hCG. The profiles of miRs showed divergent changes between oocytes and G-cells. CONCLUSION: miR expression profiles are altered by gonadotropins and supplementation of hCG during in vitro maturation of murine follicles. Target gene study must be necessary to validate these findings.
Key Words: MicroRNAs; Gonadotropins; In vitro culture; Mouse
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