• Title, Summary, Keyword: Maturation

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Factors Affecting In-Vitro Maturation in Porcine Oocytes (돼지난자의 체외성숙에 영향을 미치는 요인)

  • 박춘근
    • Journal of Embryo Transfer
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    • v.11 no.2
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    • pp.179-191
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    • 1996
  • In-vitro culture has provided new inforrnation on mechanisms of oocytes rnaturation and results obtained in vitro have led to new questions. In porcine, follicular and oocyte size have the crucial importance for the oocytes maturation. The addition of hormones to the culture medium was found to accelerate and facilitate meiotic maturation. The presence of some factors in serum trigger the resumption of meiosis and support the maturation of oocytes in vitro. The maturation rate of porcine oocytes was also increased by supplementation of porcine follicular fluid to the culture medium. The growth factors can stimulate nuclear maturation and enhances cytoplasnic maturation of oocytes by interaction with gonadotropins. The maturation-promoting factor brings about GVBD and the subsequent maturational events in oocytes. However, cAMP can block the spontaneous meiotic maturation of oocytes in culture. The understanding of these influences is a prerequisite to enhancing in vitro maturation of porcine oocytes.

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Effect of Hormone on In vitro Maturation of Porcine Follicular Oocytes (호르몬의 첨가가 돼지난포란의 체외성숙에 미치는 영향)

  • 임정훈;박병권;이성호;박창식
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.53-59
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    • 1997
  • This study was conducted to investigate the effect of hormones on in vitro maturation of porcine follicular oocytes. The results obtained were as follows : 1. The maturation rates of oocytes cultured in medium containing FSH 1, 10, 50 and 100$\mu\textrm{g}$/ml were 44.6, 58.5, 42.6 and 37.9%, respectively. And those were no difference to maturation rates among the medium containing FSH. However, the maturation rate(13.7%) of oocytes cultured without FSH was significantly(P<0.05) lower than those of oocytes cultured with FSH. 2. The maturation rates of oocytes cultured in medium containing hCG 0, 1, 10, 50 and 100IU/ml were 12.2, 11.9, 17.9, 21.9 and 45.6%, respectively. The maturation rate of oocytes cultured with 100IU/ml hCG was significantly(P<0.05) higher than those of oocytes cultured with 0~50IU/ml hCG concentrations. 3. The maturation rates of oocytes cultured in medium containing E2 0, 1, 10, 50 and 100$\mu\textrm{g}$/ml were 13.7, 10.5, 14.9, 11.4 and 5.9%, respectively. There were no differences to maturation rates among the E2 concentrations. 4. The FSH+hCG treatment was the highest maturation rate in medium containing different combination of FSH, hCG and E2.

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The Interaction between Epidermal Growth Factor (EGF) and Follicular Stimulating Hormone (FSH) on Nuclear Maturation of Mouse Oocytes by Using Their Inhibitor

  • Cha, Soo-Kyung;Kim, Tae-Hyung;Eum, Jin-Hee;Park, Kang-Hee;Park, Eun-A;Kim, Seung-Bum;Chung, Mi-Kyung;Lee, Dong-Ryul;Ko, Jung-Jae
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • pp.113-113
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    • 2002
  • 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.

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The Effects of Transcription / Translation Inhibitors on Meiotic Maturation of Porcine Oocyte In Vitro

  • Byun, Tae-Ho;Lee, Sung-Ho;Park, Chang-Sik;Lee, Sang-Ho
    • Proceedings of the Korean Society of Embryo Transfer Conference
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    • pp.117-117
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    • 2002
  • The oocytes from most of animal species accumulate genetic information and other necessary materials during oogenesis for the later use in the early development. Over the years oocyte maturation has been studied extensively both in vitro and in vivo. Particularly, maturation of follicular oocyte in vitro becomes one of the important tools for the studies of basic cell biology, the in vitro technology of animal production, and in particular, the somatic cell cloning by nuclear transfer. We examined meiotic maturation and cumulus expansion in the presence of translation or transcription inhibitors for varying periods of in viかo maturation (IVM) of pig oocyte. In Experiment 1, the results revealed that translation and transcription inhibitors inhibited cumulus expansion and meiotic maturation during 35h of IVM. However, 50 to 60% of the oocytes underwent nuclear maturation without cumulus expansion during 75h of IVM. The rest of the oocytes were arrested at metaphase I (40-50%) in the presence of the inhibitors. In Experiment II, the OCCs were exposed to the drugs only for 15h to examine translation and transcription inhibitors on cumulus expansion and meiotic maturation. Transcription inhibitors for 15h did not arrest meiotic maturation when the oocytes were cultured for subsequent, necessary period of IVM, whereas cumulus expansion was completely inhibited, suggesting that initial 15h is critical transcription activity far cumulus expansion. Translation inhibitors for 15h exposure did not alter cumulus expansion and meiotic maturation during subsequent culture in the absence of the drugs. In Experiment III, the OCCs were exposed to the drugs only for later 30h to examine the influence of transcription and translation inhibitors on oocyte maturation. Interestingly, all meiotic maturation underwent normally with full expansion of cumulus. Similar results were obtained from Experiment IV where 5h of exposure from 15 to 20h of IVM culture to the drugs was performed and subsequently cultured for same period in fresh medium. Taken there results together, both transcription and translation are necessary for nuclear maturation and cumulus expansion, and first 15h IVM for cumulus expansion is critical. The arrested oocytes by the drugs were still capable of undergoing nuclear maturation, although cumulus expansion was affected.

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Effects of Fetal Calf Serum and Porcine Follicular Fluid Fractionated by Gel Filtration on in vitro Maturation of Porcine Follicular Oocytes (Gel Filtration에 의해 분획된 소 태아혈청과 돼지난포액이 돼지난포란의 체외성숙에 미치는 효과)

  • 가학현;정구민;한정호;임경순
    • Korean Journal of Animal Reproduction
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    • v.19 no.4
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    • pp.251-258
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    • 1996
  • These studies were carried out to investigate the effect of gonadotropins (GTH), fetal calf serum (FCS), porcine follicular fluid (pFF) and FCS and pFF fractions obtained by the gel filtration on in vitro maturation of porcine follicular fluid. When the oocytes were cultured in TCM-199, the maturation rate was higher in pFF than in FCS in both with or without GTH and in pFF the maturation rate was higher in with GTH than in without GTH. In case of without GTH, pFF increased maturation rates in TCM-199, but not in Whitten's medium (WM). When the oocytes were cultured in WM supplemented with FCS fractions, the maturation rate(51.6%) of oocytes was significantly (P<0.05) higher in fraction B (about 30∼70 kDa) than in control, FCS and other fractions. When oocytes were cultured in WM supplemented with pFF fractions, fractions B (about 30∼70 kDa) and D (about 1∼10 kDa) were significantly (P<0.05) higher than in control, pFF and other fractions. In conclusiion, the addition of gonadotropins into the maturation media was effective for oocyte maturation. The addition of pFF was more effective than addition of FCS for maturation of porcine oocytes in vitro. And fraction B from FCS and fractions B and D from pFF was effective for oocyte maturation.

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Role of Golgi Apparatus on Regulation of Sec61β, COPG2 and Epidermal Growth Factor during Oocyte Maturation

  • Oh, Hae-In;Lee, Sang-Hee;Lee, Seunghyung;Lee, Seung Tae;Lee, Eunsong;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.39 no.2
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    • pp.37-41
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    • 2015
  • The oocyte undergoes various events during maturation and requires many substances for the maturation process. Various intracellular organelles are also involved in maturation of the oocyte. During the process glucose is essential for nuclear and cytoplasmic maturation, and adenosine triphosphate is needed for reorganization of the organelles and cytoskeleton. If mitochondrial function is lost, several developmental defects in meiotic chromosome segregation and maturation cause fertilization failure. The endoplasmic reticulum, a store for $Ca^{2+}$, releases $Ca^{2+}$ into the cytoplasm in response to various cellular signaling molecules. This event stimulates secretion of hormones, growth factors and antioxidants in oocyte during maturation. Also, oocyte nuclear maturation is stimulated by growth factors such as epidermal growth factor. This review summarizes roles of organelles with focus on the Golgi apparatus during maturation in oocyte.

Effects of Culture Conditions on The Expression of Cyclin B1 Protein during the First Meiotic Maturation in Bovine Immature Oocytes

  • Hwang Seong-Soo;Yoon Jong-Taek
    • Reproductive and Developmental Biology
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    • v.30 no.2
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    • pp.115-118
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    • 2006
  • Cyclin B1 is known to reflect the M-phase promoting factor (MPF), a universal regulator of G2/M-phase transition, activity during the process of oocytes maturation. To investigate whether culture condition affects the maturation rate and the expression of cyclin B1 protein, bovine immature oocytes are stimulated and cultured according to the following protocols: Experiment 1: denuded oocytes (denude) only, COC only, denuded oocytes+granulosa cells (denude+GCs) and COC+GCs; Experiment 2: no-activation (control), 7% ethanol for 5 min and $10{\mu}l/ml$ ionomycin for 5 min at immediately before maturation. The maturation rates of denude and no-activation group were significantly lower in both experiments (P<0.05), respectively. Co-culture or stimulation method in bovine immature oocytes culture increases the cyclin B1 expression significantly in both experiments (P<0.05). Based on these results, culture condition affects the maturation rate and the expression of cyclin B1 protein during the first meiotic maturation in bovine immature oocytes.

Maturation Induction in vitm of Rana dybowskii Oocyte by Lantlianum Ion (Lanthanum 이온에 의한 북방산개구리(Rana dybowskii) 여포난자의 성숙유도)

  • 유영란;임욱빈;권혁방
    • The Korean Journal of Zoology
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    • v.34 no.2
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    • pp.188-195
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    • 1991
  • The effect of lanthanum ion (La3 +), which is associated with the mobilization of internal calcium, on the regulation of oocyte maturation was investigated with Rana dybowskii follicles. Follicular oocytes matured (germinal vesicle breakdown, GVBD) dose dependently when they were exposed to La3+ (O.O1-1.O mM) and the maturation occurred in 9-12 hours after the la3+(0.33 mM) stimulation. lanthanum also accelerated the onset of maturation of the lollicular oocytes exhibiting spontaneous maturation. Three hours of exposure to La3+ was enough to induce the maturation. The La3 + -induced maturation was not associated with progesterone production by follicle cells, and the maturation was inhibited by forskolin (9 $\mu$ M), and cyclobeximide (0.01 - 1.0 - $\mu$g/2 ml) in the medium. The La3+ and hormone stimulated maturation showed the same patterns of protein phosphorylation and dephosphorylation during the maturation. The data suggest that the oocyte maturation by La3+ stimulation is very similar to that by progesterone. Thus, it seems that internal mobilization of Ca2+ plays a key role in the initiation of oocyte maturation in amphibia.

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Effect of Maturation Medium and Porcine Follicular Fluid on In Vitro Maturation of Porcine Oocytes (배양액 및 난포액이 돼지 난포란의 체외성숙에 미치는 영향)

  • 박병권;이규승;박창식;서길웅
    • Korean Journal of Animal Reproduction
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    • v.20 no.3
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    • pp.289-297
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    • 1996
  • This study was conducted to investigate the effects of maturaton medium and porcine follicular fluid on in vitro maturation of porcine follicular oocytes. The results obtained are as follows; 1. When the oocytes were cultured for 42 hours, the maturation rate was significantly (P<0.05) higher in TCM-HEPES(75.5%) than Ham's F-10(60.9%) and mKRB(60.7%) medium. The optimal medium for the maturation of porcine oocytes in vitro was the TCM-HEPES medium. 2. When the oocytes were cultured for 42 hours in TCM-HEPES medium with 10, 20 and 30% of porcine follicular fluid(pFF), the maturation rates of porcine oocytes were 69.1, 65.6 and 63.3%, respectively. The maturation rate(51.4%) of oocytes cultured without pFF was significantly(P<0.05) lower than that of oocytes cultured with pFF. 3. The maturation rates of porcine oocytes cultured for 42 hours in TCM-HEPEs medium with 3 different porcine follicular fluid treatments were 68.6%(centrifused), 72.3%(filtered) and 73.1%(heat treated), respectively. The maturation rate(49.4%) of control group without pFF treatment was significantly(P<0.05) lower than that of oocytes cultured with pFF treatment.

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Effects of Fetal Bovine Serum(FBS), Calf Serum(CS) and Human Serum(HS) on In Vitro Maturation of Porcine Follicular Oocytes (FBS, CS 및 HS가 돼지난포란의 체외성숙에 미치는 영향)

  • 임정훈;한만희;서길웅
    • Korean Journal of Animal Reproduction
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    • v.21 no.4
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    • pp.381-387
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    • 1997
  • This study was conducted to investigate the effects of fetal bovine serum(FBS), calf serum(CS) and human serum(HS) on in vitro maturation of porcine follicular oocytes. The results obtained were summarized as follows : 1. The maturation rates of oocytes cultured in medium containing FBS 5, 10, 20 and 30% were 47.0, 63.5, 48.4 and 43.2%, respectively. There were significantly higher than those of non-treated group(25.3%). And the highest maturation rate was the 10% treatment. 2. The maturation rates of oocytes cultured in medium containing CS 5, 10, 20 and 30% were 55.2, 56.6, 59.4 and 46.5%, respectively. There were significantly higher than those of non-treated group(25.3%). And the highest maturation rate was the 20% treatment. 3. The maturation rates of oocytes cultured in medium containing HS 5, 10, 20 and 30% were 74.5, 78.2, 73.1 and 68.6%, respectively. There were significantly higher than those of non-treated group(29.6%). And the highest maturation rate was the 10% treatment.

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