• Journal of Embryo Transfer
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• v.33 no.4
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• pp.287-296
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• 2018

Ganglioside GM3 is known as an inhibition factor of cell differentiation and proliferation via inhibition of epidermal growth factor receptor (EGFR) phosphorylation. Our previous study showed that the exogenous ganglioside GM3 reduced the meiotic maturation of porcine oocytes and induced apoptosis at 44 h of in vitro maturation (IVM). However, the role of ganglioside GM3 in the relationship between EGFR signaling and apoptosis during porcine oocyte maturation has not yet been studied. First, porcine cumulus-oocyte complexes (COCs) were cultured in the NCSU-23 medium with exogenous ganglioside GM3 according to maturation periods (non-treated, only IVM I: 0 - 22 h, only IVM II: 22 - 44 h and IVM I & II: 0 - 44 h). We confirmed that the proportion of germinal vesicle breakdown (GVBD) increased significantly in the IVM I treated group than in the control group. We also confirmed that the meiotic maturation until M II stage and polar body formation decreased significantly in the only IVM I treated group. Cumulus cell expansion and mRNA levels of the expansion-related factors (HAS2, TNFAIP6 and PTX3) decreased significantly in the IVM I treated group than in the control group. Protein levels of EGFR, p-EGFR, ERK1/2, and p-ERK1/2 decreased significantly in the GM3-treated groups, during the IVM I period. In addition, cellular apoptosis, determined using TUNEL assay, and protein levels of Cleaved caspase 3, were increased significantly in the GM3-treated COCs during the IVM I period. Based on these results, ganglioside GM3 exposure of porcine COCs during the IVM I period reduced meiotic maturation and cumulus cell expansion via inhibition of EGFR activity in pigs.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.929-939
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• 1996

The present study was undertaken to establish a relationship between bovine follicle size and oocyte diameter, compare the nuclear maturation competence of oocytes of different diameter groups and the nuclear maturation changes in Korean Native Cattle according to in vitro maturation period. To compare the relationship between follicle size and oocyte diameter, follicles were dissected, measured, and assigned to one of the following size categories($4{\geq}mm$, 3-4mm, 2-3mm, 1-2mm, and < 1mm), investigate the maturation competence in the different-sized oocytes, which were divided into three groups( < $110{\mu}m$, 110 - < $120{\mu}m$, and ${\geq}120{\mu}m$). Oocytes were cultured in the culture medium during 0, 6, 12, 18, and 24hrs, respectively, stained, and measured the nuclear maturation degree according to period. When compared the relationship between follicle size and intrafollicular oocyte diameter, oocyte diameters of three groups of ${\geq}3mm$ follicle-sized were significantly higher than < 3mm (p<0.01). After in vitro maturation, the rates reached to MI stage of < $110{\mu}m$ oocyte groups(25%) was higher than $110-120{\mu}m$ and ${\geq}120{\mu}m$ oocyte groups(11 and 10%) reached to the same stage(p<0.01), and the rates throughout MII stage of $110-120{\mu}m$ and ${\geq}120{\mu}m$ and < $110{\mu}m$(70 and 76%) groups were higher than < $110{\mu}m$(35%)(p<0.01). When nuclear maturation rates were measured according to period, < 6hr groups(7 and 10%) showed lower rates reached to MI than ${\geq}12hr$ groups(100%), 24hr groups(76%) revealed higher rates throughout MII than 18hr groups(40%). These results indicate that the preparation of oocyte for the production of in vitro fertilization embryos and nuclear transplantation ones could be adapted, as follicle increased up to appointed size there was a corresponding increase in oocyte diameter, and differences of nuclear maturation rate revealed according to oocyte diameter and maturation period.

• Korean Journal of Animal Reproduction
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• v.23 no.3
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• pp.239-245
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• 1999

The effect of catalase on in-vitro maturation in porcine oocytes with or without cumulus cells were studied. The maturation rates were not significantly different in medium with and without catalase during in vitro maturation of oocytes. However, the maturation rates of oocytes with cumulus cells were significantly higher (P＜0.05) than in oocytes without cumulus cells regardless of the presence of catalase. On the other hand, the maturation rate in oocytes cultured with cumulus cells for 48 h (57%) was significantly (P＜0.05) higher than in oocytes with cumulus cells for first 24 h period (42%) only. In another experiment, the maturation rate was significantly (P＜0.05) higher in medium containing catalase for last 24 h period only than in medium containing catalase for first 24 h period during in vitro maturation of ooytes with or without cumulus cells. But the oocytes matured to M-II stage were observed at 24 h of culture of oocytes without cumulus cells only. When oocytes with cumulus cells were cultured for 72 h, the maturation rates was significantly (P＜0.05) higher in medium with (79%) than without (65%) catalase during in vitro maturation. These results indicate that cumulus cells are necessary for in vitro maturation of porcine oocytes, catalase have effect according to the addition periods and can prevent aging of porcine oocytes during maturation in vitro.

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.7-11
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• 2006

This study was conducted to examine the effects of hormone and sodium pyruvate on in vitro maturation of canine oocytes. Canine oocytes were collected from the ovaries of dogs and cultured in NCSU-37 medium with hormones and sodium pyruvate for 72 hr. Oocytes matured to the metaphase II (MII) stage were observed only from estradiol $17{\beta}\;(E_2)$, and the presence of gonadotropin did not improve the nuclear maturation. No oocytes were developed to the MII stage when $E_2$ was added to medium during the first 6 and 24 hrs of culture period. The presence of $E_2$ during the whole culture period enhanced the nuclear maturation to the MII stage (6.0%, P<0.05). High concentration of sodium pyruvate (2.5 mM) slightly enhanced the nuclear maturation to the metapahse I (HMI) stage, but not the MII stage. the result of the present study shows that the presence of $ E_2$ during the whole culture period of 72 hr enhances the maturation of canine oocytes to the M stage, but sodium pyruvate does not affect the nuclear maturation of the canine oocytes.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.285-296
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• 1994

Follicular oocytes were released from the graafian follicles of ovaries from 3-4 weeks old mice. The spontaenous maturation of these follicular oocyes was inhibited by the treatment of dbcAMP and progesterone and these oocytes were cultured for 2-8hr in the Modified Hank's balanced salt solution(MHBS). Ethylenediaminetetraaceticacid(EDTA) and calmoudulin antagonist, trifluoperazine (TFP) were treated to the culture medium in order to investigate whether these chemical agents inhibit calcium uptake into the oocyte and oocyte maturation. $^{45}Ca^{2+}$, 10-${\mu}$Ci/ml was added to the culture medium during the culture period. $^{45}Ca^{2+}$uptake into the oocytes was examined whether and when various kind of oocyte maturation inhibiting agents inhibit or stimulate the influx of calcium into oocytes. Dibutyryl cAMP and progesterone decrease $^{45}Ca^{2+}$uptake into the oocytes and synergistic inhibiting effect of dbcAMP and progesterone was prominent at much lower dosages. Calcium uptake into oocytes seems to be higher during first 2 hour culture period rather than next 4hr culture. After 8hr culture, calcium uptake level of the oocytes which GVBD already took place gradually approached to the level of those which were maintained at GV by the treatment of dbcAMP and progesterone. However, $^{45}Ca^{2+}$uptake into the GV maintained oocytes did not change at all even after 8hr culture period. In addition, calcium chelating agent, EDTA inhibited calcium uptake into oocytes as well as nuclear maturation of oocytes. Lower dosage used in the present study did not inhibit calcium uptake as well as oocyte maturation.

• Animal Bioscience
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• v.37 no.6
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• pp.1007-1020
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• 2024

Objective: We increased the nuclear maturation rate of antral follicle derived oocytes by using a pre-in vitro maturation (IVM) culture system and improved the developmental potential of these porcine pathenotes by supplementing with melatonin. Furthermore, we investigated the expression patterns of genes involved in cumulus expansion (HAS2, PTGS2, TNFAIP6, and PTX3) derived from small and medium antral follicles before and after oocyte maturation. Methods: Only the cumulus oocyte-complexes (COCs) derived from small antral follicles were induced with [Pre-SF(+)hCG] or without [Pre-SF(-)hCG] the addition of human chorionic gonadotropin (hCG) during the last 7 h of the pre-IVM period before undergoing the regular culture system. The mature oocytes were investigated on embryonic development after parthenogenetic activation (PA). Melatonin (10^-7 M) was supplemented during in vitro culture (IVC) to improve the developmental potential of these porcine pathenotes. Results: A pre-IVM culture system with hCG added during the last 7 h of the pre-IVM period [Pre-SF(+)hCG] effectively supported small antral follicle-derived oocytes and increased their nuclear maturation rate. The oocytes derived from medium antral follicles exhibited the highest nuclear maturation rate in a regular culture system. Compared with oocytes cultured in a regular culture system, those cultured in the pre-IVM culture system exhibited considerable overexpression of HAS2, PTGS2, and TNFAIP6. Porcine embryos treated with melatonin during IVC exhibited markedly improved quality and developmental competence after PA. Notably, melatonin supplementation during the IVM period can reduce and increase the levels of intracellular reactive oxygen species (ROS) and glutathione (GSH), respectively. Conclusion: Our findings indicate that the Pre-SF(+)hCG culture system increases the nuclear maturation rate of small antral follicle-derived oocytes and the expression of genes involved in cumulus expansion. Melatonin supplementation during IVC may improve the quality and increase the blastocyst formation rate of porcine embryos. In addition, it can reduce and increase the levels of ROS and GSH, respectively, in mature oocytes, thus affecting subsequent embryos.

• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.323-331
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• 1999

The effects of insulin-like growth factor-I (IGF-I) and cumulus cells during in vitro maturation in porcine oocytes were examined. When follicular oocytes were cultured in medium with different concentrations of IGF-I, maturation rates were 60, 61 and 62 and 72% for 0, 15 and l0ng/$m\ell$ IGF-I. In medium with 10ng/$m\ell$ IGF-I, maturation rates were not significantly difference between oocytes with (68%) and without (52%) cumulus cells during the culture. In medium with-out IGF-I, however, the maturation rates in oocytes with cumulus cells (63%) was significantly (P＜0.05) higher than oocytes without cumulus cells (32%). On the other hand, when IGF-I was added for first 24 h period or later 24 h period of culture, maturation rates were higher in oocytes with (61 and 49%) that than without (49 and 45%) cumulus cells. In experiment used medium without fetal calf serum (FCS) and porcine follicular fluid (PFF), the maturation rates in oocytes with cumulus cells for 48 h (48 and 67%) or first 24 h (46 and 63%) period after culture were significantly (P＜0.01) higher than in oocytes without cumulus cells (16 and 18%) in the presence or absence of IGF-I. These results indicated that cumulus cells is essential on maturation in vitro in porcine oocytes, but IGF-I can promote oocytes maturation of oocytes without cumulus cells in medium with FCS and PFF.

• Journal of the Korean Society of Food Culture
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• v.31 no.2
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• pp.188-193
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• 2016

The purposes of this study were to prolong the storage period and maintain quality of kimchi made from spring napa cabbage, which is used less frequently than winter napa cabbage. The results show that the firmness of kimchi from early July, the latest harvest period, was significantly higher than that of kimchi from mid-June. However, as maturation proceeded, no significant difference was observed following the harvest period when kimchi was stored for 8 weeks. Regarding pH and acidity, which are highly related to maturation of kimchi, pH generally decreased from the initial storage period while acidity increased. In the sensory evaluation, appearance was best in kimchi manufactured in late June according to manufacturing and maturation periods (p<0.05). After 4 and 8 weeks of storage, kimchi manufactured in early July had the highest scores with no significant difference. Texture and overall acceptability also showed no significant difference in each processing period.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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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.

• Journal of Environmental Science International
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• v.15 no.2
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• pp.169-175
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• 2006

The changes of contaminants and odor corresponding to anaerobic maturation process of piggery slurry were investigated by applying the additives, such as different kinds of complex microorganism products and deodorants containing microorganism activating agents. The pHs during 20-day anaerobic maturation were operated stably without great change regardless of the additives, although they were rather lower in the case that the additives were contained than the case that they were not contained. The effects of removing CODcr, $NH_3-N$, T-N, and T-S in case that the additives were not contained, were not so great during the 20-day operation and so it would be difficult to remove the organic materials and nitrogen ingredients simply with anaerobic maturation process. However, in case of anaerobic maturation process that the additives were contained, their average removal rates were improved with the values of $49\%,\;63.5\%,\;48.5\%,\;and\;30.7\%$ for above each of items, even if the 20-day of short-term maturation period was applied. Especial1y, odor intensity with the additives was lowered continuously during the operation period and it had more than two times of lowering effect compared to that without those.