• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.147-154
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• 2012

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.377-383
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• 2011

The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into $LN_2$. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol ($72.5{\pm}5.00%$, $54.88{\pm}0.66%$ and $46.00{\pm}2.40%$; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol ($34.69{\pm}4.64%$ vs $46.00{\pm}2.40%$; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: $55.81{\pm}2.94$, $55.19{\pm}3.34$ vs $47.94{\pm}3.48%$; p<0.05 and membrane integrity: $44.94{\pm}3.51$, $46.06{\pm}2.25$ vs $40.38{\pm}1.03%$; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycerol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.369-375
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• 2011

The aim of this study was to evaluate the effect of ${\alpha}$-tocopherol on blastocysts development and subsequent cryosurvival of the vitrification. The ${\alpha}$-tocopherol(0, 100, 200, 400 ${\mu}M$) was added in to culture medium for the bovine embryos. The blasocysts from the ${\alpha}$-tocopherol and untreated control groups were then frozen-thawed, and their cryosurvival was assessed by in vitro culture for 48 h. There were no differences in the overall cleavage rate($56.14{\pm}4.66$, $58.18{\pm}4.70$, $62.97{\pm}6.86$ and $51.17{\pm}7.28$) among four treatment groups. However, in blastocyst development and total cell number were significantly higher in ${\alpha}$-tocopherol 200 ${\mu}M$ ($38.60{\pm}7.12$; $106.33{\pm}3.50$) to culture medium than other treatment groups($29.30{\pm}5.24$, $31.60{\pm}7.12$ and $26.37{\pm}4.18$; $101.36{\pm}5.12$, $97.27{\pm}2.87$, and $91.23{\pm}7.52$ respectively). Before and after vitrification, the total cell number and blastocyst development of embryo were significantly higher in July to August than September to October. In conclusion, addition of ${\alpha}$-tocopherol 200 ${\mu}M$ to in vitro bovine embryo culture medium was beneficial for improving embryo quality by decreasing the embryo damage blsstocysts cell number and improving the tolerance of the embryos to cryopreservation.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.247-250
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• 2011

The objective of this study was to find out candidate genes associated with litter size trait in pigs of inbred Large Yorkshire and Landrace populations. 86 sows were screened for candidate genotypes along with litter size data recordings. Association of litter size with genotypes of candidate genes were investigated to verify the usefulness of each gene's genotypes as markers for the trait. For the lines of Large Yorkshire, PRLR3 and RBP4 genes were genotyped. Frequency distribution of PRLR3 with genotypes AA, AB and BB were each 0.14, 0.44 and 0.42. And the average litter size by PRLR3 genotypes were 8.83, 10.81 and 10.70 piglets per litter, the average estimated breeding values of which were 0.243, 0.332, 0.365, respectively for AA, AB and BB genotypes. Genotypic frequencies of RBP4 by AA, AB and BB genotypes were 0.10, 0.44 and 0.46. The average litter size by genotypes of RBP4 were 10.40, 10.57 and 10.35 piglets per litter and their corresponding average estimated breeding values were 0.451, 0.353 and 0.261, respectively for genotypes AA, AB and BB. Significance in differences among genotypes were not observed, but B allele of RBP4 seems to be associated with litter size. In Landrace lines, frequencies of RBP4 genotypes, AA, AB and BB were 0.29, 0.55 and 0.16. And the average litter size of these genotypes were 10.50, 11.08 and 11.00 piglets per litter. The corresponding averages of estimated breeding values of each genotypes were 0.172, 0.135 and 0.104. In Landrace lines, allele A was more likely to be associated with litter size, even if differences among average litter size were not significant. We conclude that genotyping of two candidate genes is a helpful tool to identify genetic potentials of litter size in pigs.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.199-206
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• 2012

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol ($39.85%{\pm}11.41$) than in 5% glycerol treatment ($18.08%{\pm}1.61$). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol ($34.12%{\pm}11.02$) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

• Journal of Embryo Transfer
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• v.21 no.4
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• pp.299-306
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• 2006

This study examined the effects of the in vitro produced (IVP) Hanwoo blastocyst stage (blastocyst, expanded blastocyst and hatched blastocyst), in vitro culture day (7, 8, and 9) and blastocyst grade (1, 2 and 3) on the pregnancy rate, gestation length, birth weight, the incidence of dystocia and twining rate after embryo transfer (ET). The pregnancy and abortion rates were significantly higher in the blastocyst (B) stage (64.4%) and in the hatched blastocyst (HB) stage (21.4%), respectively, than in those of the other developmental stages (p<0.05). The pregnancy rate of Day 7 embryos (49.0%) was significantly higher than those of Days 8 and 9 embryos (36.4 and 15.4%), but the abortion rates were similar (0 to 10.7%). There were no significant differences in the pregnancy (41.4 to 42.5%) and abortion (9.3 to 16.5%) rates among the three grades of embryos. There were no significant differences in gestation length, birth weight and the incidence of dystocia among the three development stages, but the twinning rate was significantly higher in the HB stage (p<0.05). The pregnancy rate, the incidence rate of dystocia and twinning rate were similar among the three different culture days, however birth weight was significantly heavier in calves from Day 9 embryos than in those from Days 7 and 8 embryos. The mean gestation length of grades 1 and 2 embryos (278.5 and 276.1 days) were significantly longer than that of grade 3 (p<0.05), but birth weight, the incidence of dystocia and twinning rate did not significantly differ. The mean gestation length in single calves was significantly longer than that in twin calves (278.5 vs. 272.5 days, p<0.05). In addition, the mean birth weight in single calves was significantly greater than that in twin calves (29.6 vs. 22.3 kg, p<0.05). Finally, the sex ratios and mean mortality rates between single and twin calves were similar.

• Clinical and Experimental Pediatrics
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• v.51 no.3
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• pp.315-322
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• 2008

Purpose : Prader-Willi syndrome (PWS) is a complex genetic disorder, caused by the deletion of the paternally derived 15q11-13 region or the maternal uniparental disomy of chromosome 15 (mUPD(15)). In this study, we compared phenotypic differences between those patients whose disease was caused by microdeletion and those caused by mUPD(15). In addition, a comparison of the efficacy of growth hormone (GH) therapy between these two PWS genotypes was analyzed. Methods : Fifty-three patients were diagnosed as having PWS based on molecular and cytogenetic analyses and clinical features. Data that included maternal age, birth weight, a feeding problem in the neonatal period, cryptorchidism, developmental delay or mental retardation, short stature, hypopigmentation, changes in height, weight, and body mass indexes (BMI) before and after GH treatment were obtained by a retrospective review of medical records. The data from the patients with microdeletion were compared with those from the patients with mUPD(15). Results : Of the 53 patients with genetically confirmed PWS, 39 cases had microdeletion and 14 mUPD(15). Maternal ages were significantly higher in the mUPD(15) group, and hypopigmentation and a feeding problem in the neonatal period were more frequent in the microdeletion group. Growth hormone was administered to 20 patients [14 with microdeletion, 6 with mUPD(15)]. There were no differences between the two groups in height velocity, weight and height SDS, and BMI after GH therapy. Conclusion : Phenotype and genotype correlations were observed in Korean PWS patients, such as more advanced maternal ages in the mUPD(15) group and more feeding problems and hypopigmentations in the microdeletion group. Further long-term prospective studies are needed to correlate other aspects of the phenotypes.

• Korean Journal of Animal Reproduction
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• v.11 no.3
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• pp.196-205
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• 1987

This study was conducted in order to obtain the informatin of the histochemical changes in each of 6 segments of the epididymal ducts in 32 Korean native male goats. The male goats were examined, dividing into 7 groups, at 4 잔 intervlas from 8 to 32 wks of age. The reuslts obtained were as follows: 1. PAS reaction showed positive on the basal and upper part beyond the nucleus of the peithelium of effernt ductules throughout all the classes of age. It was also positive on the free border and basal and upper part beyond the nucleus of the caput, on the free border andbasal parts of the corpus, and on the basal part of the cauda of the epididymal epithelium. 2. Acid phosphatase reaction was negative on the every part of epididymal epithelium at the age of 8 weeks, however, with the aging it became strangly positive on the areas between the free border and the nucleus, and moderately positive on the basal part of epithelium of the caput and corpus. In the free border adn basal part of the cauda, it was slightly positive. alkaline phosphatase reactin was negative on the every part of epididymal epithelium until 12 weeks of age. From 16 weeks, free border of epididymal epithelium becaqme slightly positive, and from 20 weeks, the reaction became stronger on the basal part but weekend on the free border with the aging. 3. In the sudan black B staining, many blue black granules between the free border and the nucleus, some granules on the basal part, and a few granules on the cytoplasm around the nucleus of the epididymal epithelium were observed from 8 weeks of age as early, and the granules were increased in number with the aging. 4. In Azan staining, reddish violet granules below the nucleus and blue granules on the upper part beyond the nucleus in some cells of epithelia of efferent ductules were noted at 12th and 16th week, and after 24th week, the granules were decreased with the aging. Golgi apparatus were clearly observable on the upper part beyond the nucleus of all parts of epididymal epithelium from 8th week, and also number of intracytoplasmic vacuoles(smaller ones on the upper part and larger ones on the basal part beyond the nucleus) and fine granules were increased with the aging. 5. In the toluidin blue staining, reddish purple granules on the basal part of the epithelium in all the parts of epididymal ducts, particularly brilliant in the cauda, were observed from 8th week as early. 6. In the Cowdry staining, numerous mitochondria, according to aging, were observed between the free border of epithelium and the upper part beyond the nucleus particularly in the catus and corpus of the epididymal ducts.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.119-126
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• 1996

This study was carried out to determine the optimal condition for successful and efficient c cryopreservation of zygotes, 1-cell embryos, using EFS40 which was 40% (v/v) ethylene glycol diluted in DPBS medium containing 30% Fic-oll (w/v) and 0.3 M sucrose. After mouse zygote produced by IVF was vitrified by two freezing methods, the post-warming survival rates of 1-cell zygotes were assessed as cleavage to the 2-cell stage and development into the hatching blastocysts at 5 day. In the one-step method, when embryos were directly exposed to the vitrification solution at 25$^{\circ}C$ for 1 min., survival and development rates of zygotes were 85.5% and 31.9% In the two-step method, embryos were equilibrated with a dilute 20% EG for 1, 3, 5 min. before 1 min. exposure to EFS40, re-spectively. However, the rates of development (17.7, 3.3, 0%) were lower than that of one-step method. The highest survival rate (95.9%) was obtained by one-step method which exposes embryos in EFS40 for 30 sec. In this condition, 63. 8% of cleaved 2-cell developed into hatching blastocysts. In the cell number of Total and ICM using differential labelling technique, there are no significant differences in the cell number of Total and ICM between blastocysts devel oped in vitrified-thawed embryos (63.2${\pm}$16.9, 1 13.5${\pm}$4.0) and control balstocysts (54.0${\pm}$15.2, 1 12.3${\pm}$4.6). Therefore, these results show that mouse zygotes can be successfully cryopreserved by a simple vitrification method although developmental rates of vitrified embryos were reduced. In conclusion, this proposed vitrifi cation procedures can be useful in the cryopreservation of mouse IVF zygotes.

• Reproductive and Developmental Biology
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• v.37 no.1
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• pp.23-27
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• 2013

This study was carried out to investigate the influence of artificial insemination (AI) failure in 1,619 Korean native cows at Gangwon East area, Korea. The average AI failure rate was 37.02% in the cows and the highest rate was 40.85% in Yangyang-city. Based on the parity in the cows, the AI failure rate was 49.14% and 29.91% in the first and fifth parity cows, respectively. Whereas cows until fifth parity were decreased in AI failure, cows with sixth or more parity showed an upturning AI failure trends with the increasing of parity number. AI failure rate incidence according to the rump fat thickness measured by ultrasound was 28.9% and 33.4% at 5 mm to 10 mm and over than 15 mm, respectively. There was a positive correlation (0.2186) between AI failure rate of mother and that of their offspring cows. That is, offspring of dams with high AI failure rate showed also higher AI failure than those of dams having lower AI failure rate. In conclusion, these results indicate that the AI failure rate was closely related to the rump fat thickness, parity number, and conception rate of mother cows. In addition, these results might strengthen the basis to improve the reproductive performance in Korean native cows.