• Korean Journal of Environmental Biology
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• v.38 no.2
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• pp.323-331
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• 2020

The method of natural spawning is very passive and inconvenient for the study of developmental engineering in marine medaka, Oryzias dancena. The optimum concentration of human chorionic gonadotropin (HCG) and carp pituitary extract (CPE) for ovulation and spawning, and the injection time for the artificial spawning of marine medaka were analyzed in this study. The success rate, survival rate, and hatching rate were highest with 100IU HCG kg^-1 BW and 5mg CPE L^-1 in both male and female marine medaka (p<0.05). After obtaining unfertilized eggs and sperm by the injection of HCG and CPE into the broodstock of marine medaka, artificial fertilization could be successfully achieved any time fertilized eggs are needed in this species. This result should be useful for developing a study program for marine medaka as an experimental animal.

• Korean Journal of Veterinary Research
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• v.36 no.1
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• pp.235-245
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• 1996

Serial ultrasonographic examinations were performed on 9 pregnant Korea Jin-do bitches that were the Korean native breed, from days 15 to 60 pregnancy to determine the time of first detection and ultrasonographic appearance of the fetal and extra-fetal structures of pregnancy. Gestational age was timed from the day of ovulation (Day 0), which was estimated to occur when plasma progesterone concentration was first increased above 4.0 ng/ml. Gestational ages at earliest detection of the following fetal and extra-fetal structures were; gestational sac at days 17 to 22; placental layers in the uterine wall at days 20 to 24; zonary placenta at days 25 to 28; yolk sac membrane at days 22 to 24; amnionic membrane at days 27 to 29; embryo initial detection at days 21 to 23; fetal heartbeat at days 21 m 25; bipolar shape embryo at days 25 to 26; fetal movement at days 28 to 31; limb buds at days 31 to 35; anechoic area in head at days 31 to 36; stomach at days 34 to 37; urinary bladder at days 34 to 37; skeleton at days 36 to 38; dorsal sagittal tubular structure in vertebrae at days 36 to 38; lung hyperechoic vs liver at days 37 to 39; liver hypoechoic vs abdomen at days 37 to 40 and kidney at days 43 to 48, respectively.

• Korean Journal of Ichthyology
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• v.21 no.3
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• pp.141-148
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• 2009

Sex differentiation and gonad development were investigated in a marine medaka species, Oryzias dancena (Beloniformes; Teleostei). The average time to hatch was 11 days post-fertilization (dpf) at $25^{\circ}C$. Primordial germ cell (PGC) was first observed at 5 dpf and migrated to presumptive gonadal area between the gut and pronephric duct at 9 dpf. Male and female gonads were morphologically differentiated at 12 days post-hatching (dph). Early oocytes at perinucleolus stage as well as the formation of spermatid and efferent duct were observed at 28 dph. At 6 weeks of age, the ovary exhibited yolk granulation in many oocytes, while testis possessed a considerable number of spermatogonia and spermatids. The first ovulation was observed in 9-week-old females, and at the same age, males contained fully-matured spermatozoa. Data obtained in this study indicate that the gonad differentiation of O. dancena is the typical type of differentiated gonochorism.

• Korean Journal of Ichthyology
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• v.21 no.4
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• pp.227-238
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• 2009

The egg development and morphological changes of larvae, juveniles and adults of Oryzias dancena were observed. Fertilized eggs were incubated at $25{\pm}1^{\circ}C$; the process of embryonic development was observed by light microscopy and based on diagnostic features of the developing embryos. The average time to hatch was 11 days after fertilization. The hatched larvae averaged $4.40{\pm}0.24mm$ in total length (TL). The yolk sacs of the larvae were almost absorbed at 3 days after hatching and $4.55{\pm}0.23mm$ TL. At 21 days after hatching, the larvae were $7.23{\pm}0.73mm$ TL and had reached the juvenile stage. First ovulation was about 9 weeks after hatching and at $22.58{\pm}2.73mm$ TL.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.237-244
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• 2011

Ski protein is implicated in proliferation/differentiation in a variety of cells. We had previously reported that Ski protein is present in granulosa cells of atretic follicles, but not in preovulatory follicles, suggesting that Ski has a role in apoptosis of granulosa cells. The alternative fate of granulosa cells other than apoptosis is to differentiate to luteal cells, however, it is unknown whether Ski is expressed and has a role in granulosa cells undergoing luteinization. Thus, the aim of the present study was to examine whether the initiation of luteinization with luteinizing hormone (LH) directly regulates expression of Ski in the luteinized granulosa and luteal cells after ovulation by in vitro models. RT-PCR and real time PCR analysis respectively revealed that LH had no effect on c-Ski mRNA expression in the cultured granulosa cells regardless of LH treatment. Though Ski protein is absent in granulosa cells of preovulatory follicle, its mRNA (c-Ski) was expressed and the level was unchanged even after LH surge. Taken together, these results demonstrated that Ski protein expression is induced in granulosa cells upon luteinization, and suggested that its expression is regulated post-transcriptionally. Moreover, expression of mRNA of Arkadia, an E3 ubiquitin ligases, in luteinizing granulosa cells in vivo was assessed by realtime-PCR. The levels of Arkadia mRNA expression were unchanged during follicular growth and postovulatory luteinization. These findings suggest that Ski protein level may be regulated during luteinization at translational and/or post-translational level but not by Arkadia.

• Journal of Embryo Transfer
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• v.26 no.1
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• pp.13-17
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• 2011

Research in the area of equine artificial insemination (AI) has led to its increased application in field trials. However, procedures for equine semen collection, cooling and freezing of semen and artificial insemination need further improvement. In experiment 1, we investigated the percentage of total motility (TM) and progressive motility (PM) of sperms at after-collection, cooled-diluted, cooled-transported or frozen-thawed semen. In experiment 2, mares were inseminated with either cooled-diluted, cooled-transported or frozen-thawed semen. In experiment 3, we examined the effect of buffer (skim-milk extender), which was infused into the uterus at the time of AI with frozen-thawed semen. In experiment 4, we compared AI pregnancy rates for mares ovulating spontaneously versus after treatment with hCG. In experiment 1, the average percentage of TM was decreased from 75.3% to 14.4% at the stage of after-collection to frozen-thawed semen (p<0.05). The average percentage of PM was 58.2% and 59.6% at after-collection and cooled-diluted, but it was significantly increased 71.7% after frozen-thawed (p<0.05). In experiment 2, the pregnancy rates after AI using cooled-diluted, cooled-transported and frozen-thawed semen were 60%, 50% and 37.5%, respectively, and similar among treatments. In experiment 3, the pregnancy rate of mares infused with buffer at AI was 40% which was higher than that with no buffer (10%). In experiment 4, the pregnancy rates of mares were similar between ovulated spontaneously (25%) and ovulated with hCG (50%). The results suggest that equine semen that has been cooled-diluted, cooled-transported or frozen can be successfully used to establish AI, pregnancy and foal production. Also, the pregnancy rates after AI can be increased by infusing buffer into the uterus at AI or by inducing ovulation with hCG, but further study is need.

• Korean Journal of Veterinary Research
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• v.30 no.3
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• pp.349-354
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• 1990

The effects of $PGF2{\alpha}$ on the conception rate and the plasma levels of estradiol-$17{\beta}$ and progesterone of anestrus Cheju mares were investigated at the breeding and non-breeding seasons. The results obtained from this studies are as follows; 1. The durations of the estrus and diestrus after $PGF_2{\alpha}$ treatment persisted shorter than control cycle (p<0.05), but ovulation time was fast. 2. The levels of estradiol-$17{\beta}$ and progesteron before $PGF_2{\alpha}$ treatment showed 103.8pg/ml, 8.0ng/ml in breeding season and 72.8pg/ml, 4.7ng/ml in non-breeding season, respectively. 3. The levels of estradiol-$17{\beta}$ rose to 115.4~154.0pg/ml, and 90.8~27.0pg/ml from 2nd to 6th day after the treatment of $PGF_2{\alpha}$, in breeding and non-breeding seasons, respectively, while progesterone level dropped to 1ng/ml with the sign of estrus and at 8th day rose in breeding season (p<0.05). 4. Of thirty anestrus mares investigated for $PGF_2{\alpha}$ administration, 87.5% showed estrus on an average of 3.8 days after treatment and the conception rate was 62.5% in breeding season, but the estrus and conception rates dropped 40% and 20% in non-breeding season, respectively.

• Journal of Veterinary Clinics
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• v.5 no.1
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• pp.1-7
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• 1988

An observational study was conducted to evaluate the effect of anterior pituitary gonadtropin on super-ovulation in donors, embryo quality, development stage of embryo, transfer location, corpus luteum quality of recipient, ease of transfer and induced versus natural estrus on pregnancy rates. The data were collected from 451 non-surgical transfers of fresh and frozen bovine embryos. 1. The average number of CL, formed in ovaries which were stimulated with follicular stimulating hormone, and/or APG, were 11. 1${\pm}$1.21, 12.3${\pm}$1.84, respectively. The numbers of embryos recovered were 8.2${\pm}$1.35, 8.7${\pm}$ 1.39, and the numbers of transferable embryos were 6.4${\pm}$0.98. 6.6${\pm}$1.01. respectively. 2 There was no effect of spontaneous versus prostaglandin-induced estrus of recipients on pregnany rates of either fresh (61.5% vs. 59.5%) or previously frozen(54.5% vs. 58.5%) embryos. 3. There were significant differences(p<0.05) in pregnancy rates for embryo grade 1(63.6%), grade 2 (45.4%) and grade 3 (30.4％) in fresh group and for grade-1 (60.5%), grade-2 (47.8%) and grade-3 (18.1%) in previously frozen embryos. 4. The recipients transfered with embryo morulae were found to have a lower (p<0.05) pregnancy rates than those transfered with embryos of blastocyst in be th fresh and previously frozen embryos. 5. There was no significant difference between transfer locations in pregnancy rates using both fresh and frozen embryos. 6. Attempts to select recipients for high pregnancy rates on the basis of corpus luteum (CL) quality were performed. CL were classified into 3 quality grades, on the basis of size and prominence. Quality grades 1 and 2 group had higher (p<0.05) pregnancy rates than grade-3 group using both fresh and frozen embryos. 7 Ease of transfer was ranked to a scale of one to three on the basis of increasing difficulty (time required). Transfers ranted as ease score 1 and 2 had significantly higher (P<0.05) pregnancy pregnancy rates (45.8%, 66.6%) than ease score 3 (27.7%, 31.8%) using both fresh and frozen embryos respectively.

• Clinical and Experimental Reproductive Medicine
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• v.43 no.4
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• pp.240-246
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• 2016

Objective: The study aimed to investigate the efficacy of intrauterine instillation of granulocyte colony-stimulating factor (G-CSF) on the day of ovulation triggering or oocyte retrieval in infertile women with a thin endometrium. Methods: Fifty women whose endometrial thickness (EMT) was ${\leq}8mm$ at the time of triggering during at least one previous in vitro fertilization (IVF) cycle and an index IVF cycle were selected. On the day of triggering (n = 12) or oocyte retrieval (n = 38), $300{\mu}g$ of G-CSF was instilled into the uterine cavity. Results: In the 50 index IVF cycles, the mean EMT was $7.2{\pm}0.6mm$ on the triggering day and increased to $8.5{\pm}1.5mm$ on the embryo transfer day (p< 0.001). The overall clinical pregnancy rate was 22.0%, the implantation rate was 15.9%, and the ongoing pregnancy rate was 20%. The clinical pregnancy rate (41.7% vs. 15.8%), the implantation rate (26.7% vs. 11.7%), and the ongoing pregnancy rate (41.7% vs. 13.2%) were higher when G-CSF was instilled on the triggering day than when it was instilled on the retrieval day, although this tendency was likewise not statistically significant. Aspects of the stimulation process and mean changes in EMT were similar in women who became pregnant and women who did not. Conclusion: Intrauterine instillation of G-CSF enhanced endometrial development and resulted in an acceptable pregnancy rate. Instillation of G-CSF on the triggering day showed better outcomes. G-CSF instillation should be considered as a strategy for inducing endometrial growth and good pregnancy results in infertile women with a thin endometrium.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.159-163
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• 2017

The artificial insemination (AI) is one of the best assisted reproductive technologies for increasing reproductive capacity and facilitating the genetic improvement in farm animals. AI has been used in Uganda for over 60 years, but a small population of the total herd has been used. This study was conducted to investigate the efficacy of AI with estrus synchronization technique and to propose ways of improving the productivity of dairy farms through AI services in Uganda. In total, 78 cows from 11 dairy farms were selected for timed-AI. Synchronization was performed according to the ovsynch programs followed by AI using frozen semen from Korean Holstein (0.5 ml straws). Pregnancy rate was varying among farms (0-50%) and the overall pregnancy rate was 28.2%. Cows in luteal phase at the time of treatment was 40.0% whereas that in follicular phase was 20.8%. After treatment, cows that showed normal estrus signal were 45.5% (25/55). Abnormal estrus was categorized into pre-estrus (9.1%), cystic ovaries (21.8%), anestrus (18.2%) and delayed ovulation (5.5%), respectively. These results imply that an assured protocol for timed-AI should be developed to improve the productivity of dairy farms through AI services in Uganda.