Treatment of Epidermal Growth Factor (EGF) enhances Nuclear Maturation of Porcine Oocytes and Stimulates Expression of ER/Golgi Transport Proteins
1
조회
0
다운로드
0
인용
0
참고문헌
키워드
초록
This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear matu-ration and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 sub-unit beta (Sec61β), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte com-plexes 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 immuno-fluorescence. 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 im-portant 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.
Abstract
This study was conducted to investigate stimulatory effect of epidermal growth factor (EGF) on nuclear matu-ration and the expression level of EGF-receptor (EGFR), GM-130 (a marker of Golgi apparatus), transport protein Sec61 sub-unit beta (Sec61β), and coatomer protein complex subunit gamma 2 (COPG2) in porcine oocytes. The cumulus-oocyte com-plexes 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 immuno-fluorescence. 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 im-portant 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.
참고문헌 (28)
- Bolamba D, "Effects of epidermal growth factor and hormones on granulosa expansion and nuclear matura-tion of dog oocytes in vitro", Theriogenology, 65, pp.1037~1047 (2006)
- Conti M, "Role of the epidermal growth factor network in ovarian follicles", Mol Endocrinol, 20, pp.715~723 (2006)
- Coskun S, "Mechanism of action of epidermal growth factor-induced porcine oocyte maturation", Mol Reprod Dev, 42, pp.311~317 (1995)
- Cui XS, "Transcription profile in mouse four-cell, morula, and blastocyst: Genes implicated in compaction and blastocoel formation", Mol Reprod Dev, 74, pp.133~143 (2007)
- De La Fuente R, "Epidermal growth factor enhances preimplantation developmental competence of maturing mouse oocytes", Hum Reprod, 14, pp.3060~3068 (1999)
- De Prada JKN, "Role for cumulus cell-produced EGFlike ligands during primate oocyte maturation in vitro", Am J Physiol Endocrinol Metab, 296, pp.E1049~E1058 (2009)
- Eppig JJ, "Coordination of nuclear and cytoplasmic oocyte maturation in eutherian mammals", Reprod Fertil Dev, 8, pp.485~489 (1996)
- Eppig JJ, "Relationship between the developmental programs controlling nuclear and cytoplasmic maturation of mouse oocytes", Dev Biol, 164, pp.1~9 (1994)
- Filardo EJ, "Estrogen action via the G protein-coupled receptor, GPR30: stimulation of adenylyl cyclase and cAMPmediated attenuation of the epidermal growth factor receptor-to-MAPK signaling axis", Mol Endocrinol, 16, pp.70~84 (2002)
- Gall L, "EGFinduced EGF-receptor and MAP kinase phosphorylation in goat cumulus cells during in vitro maturation", Mol Reprod Dev, 71, pp.489~494 (2005)
- Gall L, "Expression of epidermal growth factor receptor in the goat cumulus-oocyte complex", Mol Reprod Dev, 67, pp.439~445 (2004)
- Greenfield JA, "The Sec61 complex is located in both the ER and the ER-Golgi intermediate compartment", J Cell Sci, 112, pp.1477~1486 (1999)
- Lindbloom S, "Potential involvement of EGF-like growth factors and phosphodiesterases in initiation of equine oocyte maturation", Anim Reprod Sci, 103, pp.187~192 (2008)
- Lonergan P, "Role of epidermal growth factor in bovine oocyte maturation and preimplantation embryo development in vitro", Biol Reprod, 54, pp.1420~1429 (1996)
- Mao L, "Behaviour of cytoplasmic organelles and cytoskeleton during oocyte maturation", Reprod Biomed Online, 28, pp.284~299 (2014)
- Mattioli M, "Effect of follicle somatic cells during pig oocyte maturation on egg penetrability and male pronucleus formation", Gamete Res, 20, pp.177~183 (1988)
- Mendis C, "Effect of 5-lipoxygenase inhibitor MK591 on early molecular and signaling events induced by staphylococcal enterotoxin B in human peripheral blood mononuclear cells", FEBS J, 275, pp.3088~3098 (2008)
- Moreno RD, "Golgi apparatus dynamics during mouse oocyte in vitro maturation: effect of the membrane trafficking inhibitor brefeldin", Biol Reprod, 66, pp.1259~1266 (2002)
- Payne C, "Golgi dynamics during meiosis are distinct from mitosis and are coupled to endoplasmic reticulum dynamics until fertilization", Dev Biol, 264, pp.50~63 (2003)
- Procházka R, "Developmental regulation of effect of epidermal growth factor on porcine oocyte-cumulus cell complexes: Nuclear maturation, expansion, and F‐actin remodeling", Mol Reprod Dev, 56, pp.63~73 (2000)
- Ripamonte P, "Differential gene expression and developmental competence in in vitro produced bovine embryos", Zygote, 20, pp.281~290 (2011)
- Samuelson JC, "YidC mediates membrane protein insertion in bacteria", Nature, 406, pp.637~641 (2000)
- Shimada M, "Paracrine and autocrine regulation of epidermal growth factor-like factors in cumulus oocyte complexes and granulosa cells: key roles for prostaglandin synthase 2 and progesterone receptor", Mol Endocrinol, 20, pp.1352~1365 (2006)
- Steeves T, "Metabolism of glucose, pyruvate, and glutamine during the maturation of oocytes derived from pre‐pubertal and adult cows", Mol Reprod Dev, 54, pp.92~101 (1999)
- Sutton-McDowall ML, "Effect of hexoses and gonadotrophin supplementation on bovine oocyte nuclear maturation during in vitro maturation in a synthetic follicle fluid medium", Reprod Fertil Dev, 17, pp.407~415 (2005)
- Uhm SJ, "Epidermal growth factor can be used in lieu of follicle-stimulating hormone for nuclear maturation of porcine oocytes in vitro", Theriogenology, 73, pp.1024~1036 (2010)
- Wang YN, "The translocon Sec61β localized in the inner nuclear membrane transports membrane-embedded EGF receptor to the nucleus", J Biol Chem, 285, pp.38720~38729 (2010)
- Yarden Y, "The EGFR family and its ligands in human cancer: signalling mechanisms and therapeutic opportunities", Eur J Cancer, 37, pp.3~8 (2001)