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대한생식의학회지 제32권 제3호 2010년
체외배양 중인 생쥐 난소에서 초기난포 조절인자의 발현
차병원 여성의학연구소1, 포천중문 의과대학교 생명과학대학원2
윤세진1, ,김기령2,정형민1,2,윤태기1,차광렬1,2,이경아1,2
mRNA Expression of the Regulatory Factors for the Early Folliculogenesis in vitro
, Ki-Ryeong Kim2, Hyung-Min Chung1,2, Tae-Ki Yoon1,Kwang-Yul Cha1,2, Kyung-Ah Lee1,2
1Infertility Medical Center, CHA General Hospital, 2Graduate School of Life Science and Biotechnology, Pochon CHA University College of Medicine, Seoul, Korea
.
Objective: To understand the crucial requirement for the normal early folliculogenesis, we evaluated molecular as well as physiological differences during in vitro ovarian culture. Among the important regulators for follicle development, anti-Müllerian hormone (AMH) and FSH Receptor (FSHR) have been known to be expressed in the cuboidal granulosa cells. Meanwhile, it is known that c-kit is germ cell-specific and GDF-9 is also oocyte-specific regulator. To evaluate the functional requirement for the competence of normal follicular development, we investigated the differential mRNA expression of several factors secreted from granulosa cells and oocytes between in vivo and in vitro developed ovaries.
Materials and Methods: Ovaries from ICR neonates (the day of birth) were cultured for 4 days (for primordial to primary transition) or 8 days (for secondary follicle formation) in α-MEM glutamax supplemented with 3 mg/ml BSA without serum or growth factors. The mRNA levels of the several factors were investigated by quantitative real-time PCR analysis. Freshly isolated 0-, 4-, and 8-day-old ovaries were used as control.
Results: The mRNA of AMH and FSHR as granulosa cell factors was highly increased according to the ovarian development in both of 4- and 8-day-old control. However, the mRNA expression was not induced in both of 4- and 8-day in vitro cultured ovaries. The mRNA expression of GDF-9 known to regulate follicle growth as an oocyte factor was different between in vivo and in vitro developed ovaries. In addition, the transcript of GDF-9 was expressed in the primordial follicles of mouse ovaries. The mRNA expression of c-kit was not significantly different during the early folliculogenesis in vitro.
Conclusion: This is the first report regarding endogenous AMH and FSHR expression during the early folliculogenesis in vitro. In conclusion, it will be very valuable to evaluate cuboidal granulosa cell factors as functional marker(s) for normal early folliculogenesis in vitro.
키워드 : Early folliculogenesis, Ovarian culture, AMH, FSH Receptor, GDF-9, c-kit
교신저자 : leeka@ovary.co.kr
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