The objective of this study was to examine the effect of EGF on meiotic maturation and pronuclear (PN) formation of porcine oocytes. Prepubertal gilt cumulus-oocyte-complexes (COCs) aspirated from 2~6mm follicles of abbatoir ovaries were matured in TCM199 containing 0.1mg/ml cysteine, 0.5㎍/ml FSH and LH, and EGF (0, 5, 10, 20, 40 ng/ml) for 22 hr at 39℃ in a humidified atmosphere of 5% CO2 in air. They were then cultured for an additional 22hr without hormones. In Experiment 1, to examine the nuclear maturation at 44hr of culture, the expanded cumulus cells were removed by vortexing for 1 min in 3 mg/ml hyaluronidase. The oocytes were fixed in acetic acid: methanol (1:3, v/v) at least for 48 hr and stained with 1% orcein solution for 5 min. Nuclear status was classified as germinal vesicle (GV), germinal vesicle breakdown (GVBD), prophase-metaphase I (PI-MI), and PII-MII under microscope. In Experiment 2, to investigate PN formation, oocytes were fertilized with Percoll-treated freshly ejaculated sperm (1 x10 5 cells/ml) in mTBM with 0.3% BSA and 2mM caffeine for 5hr, and cultured in NCSU-23 medium with 0.4% BSA. At 6hr of culture, the embryos were fixed in 3.7% formaldehyde for 48hr and stained with 10ug/ml propidium iodide for 30 min. PN status was classified as no or one PN (unfertilized), 2 PN (normal fertilized) and ≥3 PN (polyspermy). Differences between groups were analyzed using one-way ANOVA after arc-sine transformation of the proportional data. The rate of oocytes that had reached to PII-MII were significantly (P<0.05) higher in all groups added EGF than that of non-treated group (67%), but it did not differ among the all added groups (86%, 85%, 79% and 81%, in 5, 10, 20 and 40 ng/ml EGF, respectively). No differences on the incidence of 2PN were observed in all treated groups (25%, 30%, 33%, 29% and 29%, in 0, 5, 10, 20 and 40 ng/ml EGF, respectively), however, in non-treated group, polyspermy tended to be increased (66% vs 58%, 54%, 52% and 55%, 0 vs. 5, 10, 20, 40 ng/ml EGF, respectively). These results suggest that EGF can be effectively used as an additive for enhancing oocyte maturation and reducing the incidence of polyspermy in pig.

The stimulatory effect of EGF and FSH on oocyte maturation have been reported in various mammalian species. And some reports presented FSH enhanced the effect of EGF on oocyte maturation. But, the interaction between EGF and FSH on nuclear maturation of mammalian oocytes is not fully understood. We observed the effect of EGF and FSH on nuclear maturation during in vitro maturation of mouse oocytes. Also, we examined the interaction between EGF and FSH on nuclear maturation of mouse oocytes using the EGFR inhibitor or FSH inhibitor. Germinal vesicle (GV) stage oocytes were obtained from 3-4weeks PMSG primed BCFI hybrid mice and cultured in TCM-199 medium with 0.4%PVP supplemented with/without EGF (1ng/ml), FSH (1ug/ml), EGFR specific tyrosine kinase inhibitors: Tyrphostin AG 1478 (500nM), MAP kinase kinase inhibitor : U0126 (20uM) or PD 98059 (100uM) for 14-l5hr. Rapid staining method were used for the assessment of nuclear maturation. Nuclear maturation rates of EGF indjor FSH-treated group were significantly higher than those of control group. Treatment of EGFR inhibitor significantly block the nuclear maturation of GV oocyte in EGF-treated group, but it did not block those of GV oocyte in FSH-treated or FSH and EGF-treated group. Treatment of FSH inhibitor(U0126, PD98059) significantly block the nuclear maturation of EGF-treated group, FSH-treated and FSH and EGF-treated group. These results show that EGF has a stimulatory effect as well as different action pathway with FSH on in-vitro maturation of mouse oocyte in vitro. Therefore, further studies will be needed to find the signaling pathway of EGF associated with nuclear maturation.

Heparin-bindin epidermal growth factor (HB-EGF) is one of the EGF family to be expressed at the time of implantation in the mouse uterus. Although HB-EGF has been shown to stimulate the development of embryo and uterus in the mouse, its correlation between cell adhesion molecules remains undefined. Integrin , one of the cell adhesion molecules, is an important mediator of cell-substratum and cell-cell adhesion in implantation. In the present studies, we investigated the effects of HB-EGF on the embryonic development, initiation of implantation and expression of integrin in in vitro culture, blocking of HB-EGF, RT-PCR and immunofluores cence analysis. The results showed that HB-EGF significantly improved the developmental rate of hatched embryos (24.1%, p<0.01) and outgrowth embryos (42.5%, p<0.01). On the other hand, this growth factor showed no offset before the hatching embryonic stage. Analysis of RT-PCR showed that HB-EGF upregulated the expression level of integrina subunit genes on the preimplantation embryo and outgrowth of blastocyst (120hr and 144hr after hCG injection). Immunofluorescence analysis showed that the integrin subunits localized at the pericellular borders and cell-cell contact areas. Increase in fluorescence intensity was observed in the HB-EGF treated embryos. Intrauterine injection of an anti-HB-EGF antiserum at day 3 significantly decreased the number of implantation sites (14.4, p<0.01) and significantly increased the number of recovered embryos(6.4, p<0.05) at day 5. From these results, it imply that HB-EGF improve the embryo development and accelerated the expression of integrin in the preimplantation mouse embryos.

본 연구는 성장인자 EGF, IGF-1, EGF+IGF-1 이 소 난포란의 체외성숙에 미치는 영향을 규명하기 위하여 실시하였다. 소 난포란의 체외성숙 시 EGF를 농도별 (10, 50 및 100ng/m1)로 첨가하여 실험한 결과 통계학적 유의성은 인정되지 않았으나 24시간 배양 후 성숙율은 대조군에 비하여 높은 경향을 나타내었다. 소난포란의 체외성숙 시 EGF와 IGF-1을 병용 처리하여 실험한 결과, EGF 또는 IGF-1 단독처리 군에 비하여 병용처

This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear maturation and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 subunit beta (Sec61β), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte complexes were collected from follicle with 3-6 mm in diameter. They were incubated in medium with/without EGF for 22 h (IVMⅠ) and subsequently incubated hormone-free medium with/without EGF for 22 h (IVMⅡ). Nuclear maturation state was checked by aceto-orcein stain. Protein expression of EGFR, GM-130, Sec61β, and COPG2 were measured by immunofluorescence. In results, nuclear maturation of oocytes in EGF non-treated oocytes were significantly lower than EGF-treated groups at IVMⅠ or IVMⅡ stage (P<0.05), whereas maturational rate in EGF treatment groups at both of IVM stage was higher in among the all treatment groups (P<0.05). EGFR, GM-130, Sec61β and COPG2 were expressed in the cytoplasm of oocytes. Especially, GM-130 and EGFR were strongly expressed, but Sec61β and COPG2 were weakly expressed in cortical area of cytoplasm. The protein level of GM-130, Sec61β, and COPG2 were significantly higher in the EGF-treated groups (P<0.05). However EGFR was no difference between non EGF-treated groups and control. In conclusion, EGF plays an important role in the systems for oocyte maturation with endoplasmic reticulum and Golgi apparatus. In addition, the protein levels of Sec61β and COPG2 could be changed by EGF in the porcine oocytes during maturation.

여드름은 면포, 구진, 낭종, 결절, 색소 침착 등 다양한 피부병변으로 나타나는 모낭피지선의 만성 염증질 환으로 사춘기부터 성인기까지 발생 연령대가 다양해지고 있다. 한편, 약물 부작용으로 여드름이 발생하기도 하 는데, 표피성장인자(epidermal grouwth factor, EGF) 수용체 억제제 항암제를 사용할 경우 75 ∼ 100%에서 여드름양 모낭염이 발생된다고 보고되고 있다. 여드름의 치료로 항생제, 레티노이드 경구 복용 및 외용 약제 도 포 등 다양한 방법이 이용되고 있다. 그러나 최근 들어 레티노이드 기형 유발 가능성 및 Propionibacterium acne의 항생제 내성률 증가는 기존 치료의 한계로 여겨진다. 따라서 본 연구는 최근 여드름양 발진에 효과가 있다고 알려진 EGF를 함유한 외용제가 여드름 치료에 미치는 효과와 안정성을 평가하였다. 한국 성인 10 ∼ 29세 23명을 대상으로 EGF 함유 제품(트러블컨트롤 EGF)과 3종 제품(트러블컨트롤 클래리파잉 클렌징폼, 트 러블컨트롤 올-클리어 필링토너, 레드롤 카밍 모이스처)을 하루 두 번 사용하도록 하였다. 사용 후 영상 피지량, 경표피수분손실량, 피부 홍조 측정, 전문가 육안 평가, 사용 후 만족도 설문조사를 평가하였다. 최종 측정 시, 피부 피지량, 경피수분손실량, 피부 홍조가 통계학적으로 감소하였으며, 전문가 육안 평가에서 여드름 병변(면 포, 구진)도 통계학적으로 감소하였다. 연구동안 심각한 부작용은 관찰되지 않았다. 표피성장인자 함유 외용제는 경도의 여드름에 안전하면서도 효과적으로 사용할 수 있을 것으로 생각된다.