• Journal of the Korea Academia-Industrial cooperation Society
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• v.11 no.6
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• pp.2124-2128
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• 2010

In this study, we determined effects of the mitogen-activated protein kinase (MAPK) inhibitor, U0126 on meiotic maturation, microtubule organization and actin filament assembly in the porcine oocyte. The phosphorylated MAPK was first detected at 12 h after the initiation of maturation cultures, fully activated at 24h, and remained until metaphase II. Treatment of germinal vesicle (GV) stage oocytes with $20{\mu}M$ U0126 completely blocked MAPK phosphorylation, but germinal vesicle breakdown (GVBD) was normally proceeded. However, the oocytes didn‘t progress to the metaphase I. The inhibition of MAPK resulted in abnormal spindles. In oocytes treated with U0126 after GVBD, polar body extrusion was normal, but the organization of the metaphase plate and chromosome segregation were abnormal. In conclusion, MAPK activity plays an important regulatory role in GV chromatin configuration and meiotic progress in porcine oocyte maturation.

• The Korean Journal of Malacology
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• v.29 no.2
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• pp.97-103
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• 2013

In the present study, we monitored the early development of Saccostrea kegakia subtropical oyster species distributing on rocky intertidal off the northern Jeju Island using scanning electron microscope (SEM). The female oyster collected in early August, 2012 were fully mature exhibiting relatively small eggs ($46.5{\pm}1.4{\mu}m$ in diameter) in the gonad, while testis of the mature male oysters were filled with fully developed sperms of 36.9 ${\mu}m$ in length. The fertilized eggs developed into 2-cell stage with polar body after 1 hr 20 min of fertilization, then followed by Morula stage (3 hr 20 min), Blastula stage (4 hr 50 min), Gastrula stage (7 hr), and trochophore larvae stage (9 hr 30 min). The observed early development of S. kegaki in this study was similar the early development of other oysters, although size of the fertilized eggs were somewhat smaller.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.219-228
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• 1997

To examine the efficiency of nuclear transplantation the influence of electrical preactivation of recipient cytoplasm on the in vitro developmental potentyl in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgery procedure. The separated G1 phase blastomeres of 32-cell stage were put into the non-preactivated and/or the preactivated recipient cytoplasm by electrical stimulation. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused. The fused nuclear transplant embryos were co-cultured with rabbit oviduct epithelial cells and monitored every 24h to assess for developmental rate. After in vitro culture for 120h, the nuclear transplant embryos developed to blastocyst stage were stained with Hoechst 33342 and their blastomere were counted. The electrofusion rate was similar to the non-preactivated and preactivated recipient cytoplasm(81.8 and 85.7%, respectively). However, the in vitro developmental rate to blastocyst stage with the non-preactivated recipient cytoplasm (57.1%) was found significantly (P<0.05) higher, compared to the preactivated recipient cytoplasm(20.8%). The cell counts of nuclear transplant embryos developed to blastosyst stage were increased significantly (P<0.05) more in the non-preactivated recipient cytoplasm (163.7 cells), as compared with the preactivated recipient cytoplasm(85.4 cells). These results considered better that non-preactivated oocytes, MII phase oocytes, were used for recipient cytoplasms in the rabbit nuclear transplant procedure.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.229-238
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• 1997

Chromosome condensation and swelling of the donor nucleus have been known as the early morphological indicators of chromatin remodelling after injection of a foreign nucleus into an enucleated recipient cytoplasm. The effects of non-preactivation and electrical preactivation of recipient cytoplasm, prior to fusing a donor nucleus, on the profile of nuclear remodelling in the nuclear transplant rabbit embryos were evaluated. The embryos of 16-cell stage were collected and synchronized to G1 phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome mass by non-disruptive microsurgical procedure. The separated G1 phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation and the separated G1 phase blastomeres of 32-cell stage were injected. After culture until 20h post-hCG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The nuclei of nuclear transplant embryos fused into non-preactivated and/or preactivated recipient cytoplasm were stained by Hoechst 33342 at 0, 1.5, 2, 4, 6, 8, 10 hrs post-fusion and were observed under an fluorescence microscopy. Accurate measurements of nuclear diameter were revealed with an ocular micrometer at 200$\times$. Upon blastomere fusion into non-preactivated recipient cytoplasm, a prematurely chromosome condensation at 1.5 hrs post-fusion and nuclear swelling at 8 hrs post-fusion were occurred as 91.6% and 86.1%, respectively. But the nuclei of nuclear transplant embryos fused into preactivated recipient cytoplasm, as o, pp.sed to non-preactivated recipient cytoplasm, were not occurred chromosome condensation and extensive nuclear swelling. Nuclear diameter fused into non-preactivated and preactivated recipient cytoplasm at hrs post-fusion was 30.2$\pm$0.74 and 15.2$\pm$1.32${\mu}{\textrm}{m}$, respectively. These results indicated that onset of unclear condensation and swelling which was associated with oocytes activation were critical steps in the process of chromatin swelling. Futhermore, complete reprogramming seemed only possible after remodelling of the donor nucleus by chromosome condensation and nuclear swelling.

• Journal of Embryo Transfer
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• v.9 no.2
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• pp.153-160
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• 1994

This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 $\mu$sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39$^{\circ}C$ in a 5% $CO_2$ incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.

• Journal of the Korea Fashion and Costume Design Association
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• v.10 no.2
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• pp.39-56
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• 2008

It is a remarkable phenomenon that tattoos has been rapidly spreading among women of modern years. The extremism of feminine ornaments, and pricking and piercing of the skin seem to have deep connection with women's consciousness changing to being independent, active and aggressive. The objective of this study is to analyze the potential quality of today's women, who appears emblematically through tattoo, and to search out the whole aspect and keynote of female tattoos. The methods of research are by examining the general characteristics including concept and history, considering the type and significance of ethnic female tattoos, and then comparing with the symbolism of today's female tattoos. The sphere is limited to women's tattoos from ancient times to today's modern lift, examined from large portions of Africa, Asia, Australasia, the Americas, and Arctic regions. As a result, the following had been reflected in female tattoos in the past. First, incantation, tribe, adult age, adornment, marital status, domestic ability and sex distinction, where among others, the incantatory, ornamental symbol, and domestic sign were representatives. Second, the most popular body parts for having a tattoo were around the lips, hands and abdominal region. Third, the more harsh the environment becomes such as tropical forests, isolated islands, and polar regions, the more number of females have tatoos. Fourth, women's tattoos were much less glamorous and smaller compared to those of men, distinguishing one's sex. On the other hand, today's female tattoos can be classified as five symbols: masculinity, independence, voluptuousness, affection, and purity. These symbols contain women's mentality to pass the limit, and to overcome social inferiority. In modern society, where women must compete with met it has become necessary for women to educe their potential masculine qualities. Being violent accessories for them, tattoos became a mark of such potential. Therefore, in the future, women's interest and demand for tattoos are expected to rise in proportion to the social demand of women's force.

• Journal of Embryo Transfer
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• v.12 no.1
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• pp.1-10
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• 1997

The present study was conducted to investigate the influence of embryo cell stage at 18h post-fusion and oocyte preactivation on sebsequent in vitro developmental potential in the nuclear transplant rabbit embryos. The embryos of 16-cell stage were collected and synchronized to G$_1$ phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome rnass from the oocytes collected by non-dis-ruptive microsurgery procedure. The separated G$_1$ phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation at 18h post-hCG injection and the separated G$_1$ phase blastomeres of 32-cell stage were injected. Mter culture until 20h post-hOG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The fused nuclear transplant embryos were classified into 3~4-cell, 2-cell and 1-cell stage at 18 hrs post-fusion and cultured until the embryos reached blastocyst stage. The developmental rate to blastocyst stage was significantly (P <0.05) higher in all the reconstituted embryos of 3~4-cell stage(58.0%) than in 2 and icell stage. The developmental rate to blastocyst stage in the embryos of 3~4-cell stage at 18 hrs post-fusion was significantly (P<0.05) higher in the reconstituted without oocyte preactivation(77.8%) than in the oocyte-preactivated embryos (33.3%). These results indicated that the higher rate of in the in vitro development to blastocyst stage might be obtained form the embryos which were reconstituted with nuclear donor of G$_1$ phase and non-preactivated oocyte, and developed more rapidly for 18 hrs post-fusion.

• Journal of Veterinary Clinics
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• v.32 no.6
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• pp.536-539
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• 2015

One year old mixed-breed bitch was examined to retrieve in vivo matured oocytes. Laparotomy was performed 72 hr after ovulation determined by serum progesterone concentration, and abnormally enlarged left uterus horn was found. Both ovaries had eight corpus lutea, and a total 16 in vivo matured oocytes having perivitelline space within $25{\mu}m$, polar body, and metaphase II nucleus were recovered by flushing oviducts. This is the first study to confirm in vivo maturation of oocytes from a bitch with hydrometra, which suggests that oocytes recovered from canids with reproductive disease could be valuable sources for assisted reproductive technologies.

• The Korean Journal of Zoology
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• v.27 no.1
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• pp.1-12
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• 1984

In order to investigate the alkaline phosphtase activities in the mouse oocytes in matuation and preimplantation embryos in developing in culture, the enzyme activities were measured by means of biochemical method. The in vitro effect of levamisole which is known as an inhibitor of the lakaline phosphatase was also observed on the oocyte in maturation and the embryos in early embryogenesis. The results obtained were as follows: The enzyme activity was not detected in the embryos unitl the stage of 4-cell, but it appeared first in the 4-cell embryos and the level of the activity was steady through up to the blastocyst. Levamisole inhibited the alkaline phosphatase activity in the blastocyst, and the activity decreased by almost 70% at 10 mM and 50% at 1 mM as compared with the control. In addition, levamisole inhibited completely the formation of polar body by the oocytes. and induced degeneration of the preimplantation embryos at the dose of 0.5 mM or higher.

• 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.