• Animal Bioscience
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• v.36 no.1
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• pp.29-42
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• 2023

Objective: Pigs, an ideal biomedical model for human diseases, suffer from about 50% early embryonic and fetal death, a major cause of fertility loss worldwide. However, identifying the causal variant remains a huge challenge. This study aimed to detect single nucleotide polymorphisms (SNPs) and candidate genes for the number of mummified (NM) piglets using the imputed whole-genome sequence (WGS) and validate the potential candidate genes. Methods: The imputed WGS was introduced from genotyping-by-sequencing (GBS) using a multi-breed reference population. We performed genome-wide association studies (GWAS) for NM piglets at birth from a Landrace pig populatiGWAS peak located on SSC11: 0.10 to 7.11 Mbp (Top SNP, SSC11:1,889,658 bp; p = 9.98E-13) was identified in cyclin dependent kinase on. A total of 300 Landrace pigs were genotyped by GBS. The whole-genome variants were imputed, and 4,252,858 SNPs were obtained. Various molecular experiments were conducted to determine how the genes affected NM in pigs. Results: A strong GWAS peak located on SSC11: 0.10 to 7.11 Mbp (Top SNP, SSC11:1,889,658 bp; p = 9.98E-13) was identified in cyclin dependent kinase 8 (CDK8) gene, which plays a crucial role in embryonic retardation and lethality. Based on the molecular experiments, we found that Y-box binding protein 1 (YBX1) was a crucial transcription factor for CDK8, which mediated the effect of CDK8 in the proliferation of porcine ovarian granulosa cells via transforming growth factor beta/small mother against decapentaplegic signaling pathway, and, as a consequence, affected embryo quality, indicating that this pathway may be contributing to mummified fetal in pigs. Conclusion: A powerful imputation-based association study was performed to identify genes associated with NM in pigs. CDK8 was suggested as a functional gene for the proliferation of porcine ovarian granulosa cells, but further studies are required to determine causative mutations and the effect of loci on NM in pigs.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.81-81
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• 2003

DNA methylation is a covalent modification of DNA that can modulate gene expression and is now recognized as a major component of the epigenome. During evolution, the dinucleotide CpG has been progressively eliminated from the genome of higher eukaryotes and is present at only 5% to 10% of its predicted frequency. Approxymately 80% of the remaining CpG sites contain methylated cytosines in most vertebrates and they are distributed in a pattern that is unique in each tissue and is inversely correlated with gene expression. The pattern of methylation is faithfully maintained during cell division by the enzyme Dnmt1, the maintenance DNA methyltransferase, which catalyzes the transfer of a methyl group from S-adenosyl-methionine to the 5'-position of the cytosine ring. We have been identified bovine Dnmt1 cDNA full-length recently (AY173048) Little is known on the functions of Dnmt1 in bovine preimplantation embryos. Thus, we analyzed the specific pattern of Dnmt1 in in vitro derived/nuclear transfer bovine and in vivo derived mouse embryos to monitor the epigenetic reprogramming process. We investigated these process by using indirect immunofluresence with an antibody to Dnmt1. According to other studies, Dnmt1 accumulates in nuclei of early growing oocytes but is sequestered in the cytoplasm of mature oocytes. In 2-cell and 4-cell embryos, Dnmt1 is cytoplasmic, but at the 8-cell stage, it is present only in the nucleus. By the blastocyst stage, Dnmt1o is again found only in the cytoplasm. Thus, nuclear localization of Dnmt1o in preimplantation embryos is limited to the 8-cell stages After implantation, Dnmt1 is localized in the nucleus in mouse. However, we have found different patterns of Dnmt1 nuclear localization. Though we used the common antibody, immune-localization data revealed that Dnmt1 antibody have been detected at the nucleus in 1-cell to blastocyst embryos. Therefore, maybe we think that the functions of Dnmt1 between bovine and mice are different. In order to Identify the mechanisms that regulate DNA methylation in bovine preimplantation embryo, we have plans on using bovine oocyte and somatic specific Dnmt1 antibodies.

• Fisheries and Aquatic Sciences
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• v.27 no.4
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• pp.213-224
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• 2024

Betta rubra, classified as endangered fish species by the International Union for Conservation of Nature (IUCN), has been successfully bred and raised in captivity for two generations under laboratory conditions. This study aimed to provide comprehensive information on the captive breeding of B. rubra, focusing on various parameters crucial for ex-situ conservation and domestication. The research involved breeding trials, embryo and larvae observation, first feeding experiments, larva and fry rearing trials, and the evaluation of growth and reproduction in two generations. The study revealed that the female B. rubra, with an average total length of 5.17 ± 0.15 cm and weight of 1.61 ± 0.06 g, produced an average of 73.67 ± 7.09 eggs, 34.33 ± 5.13 total larvae, and exhibited a hatching rate of 46.67 ± 5.77%. The embryogenesis process commenced on the day of spawning (dps) and continued until the eggs hatched at 6 dps. Larvae development and yolk absorption occurred from 0 to 6 days post-hatching (dph). The study also examined the impact of different initial feeding options, with chopped Tubifex resulting in the most significant in- crease (p < 0.05) in length. The growth pattern of B. rubra larvae showed slow initial growth during the first seven days, followed by a rapid exponential growth phase from day 8 to day 39. Two generations of B. rubra (G1 and G2) were successfully bred in captivity, with G2 showing a better tendency for growth in length and weight compared to G1. Notably, there were no significant differences (p > 0.05) in reproductive success between the wild-origin broodstock (G0), G1, or G2. This research contributes valuable insights into the captive breeding of B. rubra and its early life stages, offering critical information for the conservation and sustainable management of this endangered species. Further research is needed to explore the long-term effects of domestication on behavior, physiology, and phenotypic diversity.

• Development and Reproduction
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• v.2 no.2
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• pp.179-187
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• 1998

The effects of prostaglandins in hatching and implantation have been studied but the results were various, and those are not well known by the embryonic stage. The present study examined the effects of prostaglandin $E_2$(PG $E_2$) and prostaglandin $F_2$$_{\alpha}$ (PG $F_2$$_{\alpha}$) on the expansion and hatching of mouse embryos by embryonic stage. Also we tried to measure the concentration of prostaglandins of morula, expanded, and hatching embryos. In early morula stage embryos, high concentration of PG $E_2$(>100$\mu$M) showed cytotoxicity but PG $F_2$$_{\alpha}$ did not. The hatching was inhibited all groups but not gave negative effects on expansion. In 84 hr and 96 hr stage embryos, the hatching rate was decreased at all treatment groups but not inhibited the expansion. When combine prostaglandin with indomethacin, the hatching rate was increased significantly compared to the prostaglandin-treated groups, and as lower and lower the PG $E_2$ concentration, the hatching rate increased to the control level. The embryonic synthesis of PG $E_2$ increased dramatically but that of PG $F_2$$_{\alpha}$ increased gradually. PG $E_2$ showed cytotoxicity at early stage embryos much than late stage embryos, but PG $F_2$$_{\alpha}$ did not. Hatching was inhibited by the high PG $F_2$$_{\alpha}$ concentration. It is suggested that the inhibition of hatching might be at resulted from cytotoxicity of PG $E_2$ on embryo. However, it is thought that the mechanisms of inhibition of hatching are different between PG $E_2$ and PG $F_2$$_{\alpha}$. In conclusion, it can be suggested that PG $E_2$ and PG $F_2$$_{\alpha}$ concerned with the expansion and hatching, and their effects on hatching were different by the embryonic stage.$/ concerned with the expansion and hatching, and their effects on hatching were different by the embryonic stage.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.139-146
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• 1997

This experiment was carried out to evaluate the effect of mouse early embryonic development in vitro by co-culture with bovine and porcine oviductal epithelial cells(BOEC and POEC). The 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/15% FCS at 48 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined. For a comparative study of in vi패 and in vitro development, the fresh blastocyst which developed in vivo for 120 hours after hCG injection was collected from the uterus, and their numbers of nuclei were also counted. The higher developmental rates of blastocyst formation was a, pp.ared from 91% to 97% when the embryos were co-cultured with a monolayer of bovine or porcine oviductal epithelial cells in TCM 199 or Ham's F-10 and MediCult IVF media. No significant difference in developmental rates was shown between bovine and porcine oviductal eptithelial cells. The number of nuclei in the embryos cultured for 72 hours under each conditions was significantly reduced it than blastocyst in vitro conditions. The number of nuclei in embryos cultured in TCM 199, Ham's F-10 and Medicult IVF medium were counted 68.1$\pm$6.00, 67.3$\pm$4.49, 66.4$\pm$5.64, and 94.3$\pm$8.61, 92.5$\pm$7.60, 92.1$\pm$6.10 with BOEC and 93.3$\pm$5.80, 92.9$\pm$6.53, 92.3$\pm$7.35 with POEC coculture, respectively. These numbers were lowered than 107.2$\pm$7.43 in vivo conditions. In conclusions, the coculture between the mouse early embryos, and oviductal epithelial cells of BOEC and POEC give to improve the developmental and hatching rates of blastocyst but in vivo culture systems for the growth of nuclei were ineligible than in vitro conditions.

• Korean Journal of Ichthyology
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• v.32 no.3
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• pp.166-181
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• 2020

The egg development, early life history and spawning site characters of Korean endemic fish, Coreoleuciscus splendidus (Gobioninae), were investigated at the part of Jo-jong stream in Korea from May 2020. The fertilized eggs were 2.05~2.23 mm (mean, 2.13 mm) in diameter and had no oil globules. The embryo began to hatching about 98 hrs after fertilization under water temperature of 20±1℃. The newly-hatched larvae were 5.03~5.68 mm (mean, 5.31 mm) in total length (TL), and their mouth and anus were not opened. 4 days after hatching, Several rod-like cupulae were observed on the head and lateral side of the body at 6.95~7.89 mm (mean, 7.51 mm). 7 days after hatching, the post-larva stage were 8.39~9.29 mm (mean, 8.78 mm) in total length, and their york were completely absorbed. The cupulae were completely distinguished. They entered the juvenile stage when all fin-rays were formed at 29 days after hatching, and their TL were 14.16~17.04 mm (mean, 14.99 mm). Squamation was initiated on the caudal body at approximately 18.21~23.74 mm (mean, 30.28 mm), 38 days after hatching, and completed at 26.82~33.33 mm (mean, 19.42 mm), 73 days after hatching, the external characteristics from of juveniles were same to adults. The spawning site was characterized by bottom structure of pebble (64~16 mm) and gravel (16~2 mm), environmental conditions of the spawning sites were 6~18 cm in water depth, 0.43~0.73 m/sec in bottom water velocity. In spawning sites, an egg mass or separated eggs were located a small gap of under the pebble and gravel.

• Korean Journal of Ecology and Environment
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• v.35 no.3 s.99
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• pp.198-212
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• 2002

A numerical variation and abnormalities were studied on egg bags and embryos of Korean salamander, Hynobius leechii from agricultural habitat. The teratogenic and toxic effects of fungicide benomyl were also investigated with early embryos from non-agricultural habitat. We collected 144 egg bags from agricultural region, and 3418 of early embryos were contained. The lengths of egg bags were varied from 10 to 23 cm and the most frequent length was 19 cm. The number of embryos was varied from 7 to 43, and the most frequent range was 22 to 26. Spontaneous abnormalities were occurred in 406 embryos among 116 egg bags, and 24 kinds of external abnormalities were found. Individuals showing severe external defect were histologically studied and they showed optic dyspalsia, thyroid carcinoma, somatic muscular dysplasia, partial biaxial structure, decrease of red blood cells in the heart, cephalic degeneration and intestinal dysplasia. 385 embryos from non-agricultural region were exposed to 200 nM${\sim}$ 1 ${\mu}$M of benomyl at blastula or gastrula for 12 days. All embryo were dead in the concentration of 1 ${\mu}$M (LD$_{100}$) and 75% of embryos were dead in 800nM of benomyl. Speciflc effect due to benomyl was acrania or cephalic dysplasia and this restult suggests that the benomyl inhibit stongly to the development of neural tissue. These abnormal developments may be caused by antimitotic action, inhibition of tubulin complex, destruction of microtubule, inhibitions of neurulation and closing of neural fold, and by the inhibition of the movement of neural crest cells.

• Korean Journal of Animal Reproduction
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• v.24 no.1
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• pp.89-95
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• 2000

This study was to establish an effective dilution technique in a vitrification of bovine blastocysts for the field trial. For vitrification, blastocysts were exposed in glycerol (G) and ethylene glycol (EG) mixture in m-DPBS supplemented with 10% FBS. Blastocysts were first exposed to 10% (v/v) G for 5 min, and subsequently were transferred to 10% G plus 20% EG (v/v) for 5 min. Finally, embryos were transferred to 25% G plus 25% EG (v/v) for 30 see and were placed in nitrogen vapor for 3 min, and then were plunged into L$N_2$. At thawing, the straw containing blastocysts was placed in air for 10 sec, and then plunged into a water bath at $25^{\circ}C$ until all ice had disappeared. They were placed in $25^{\circ}C$ and 36$^{\circ}C$ water according to treatment group for different time. Also, in vitro survival was assessed by the re-expansion and hatched rates at 24 hand 48 h postwarming, respectively. The results obtained in these experiments were summarized as follows; 1) In the survival rates of vitrified bovine blastocysts according to different dilution time at thawing, the data of 1 min group (86.6, 56.6%) were higher than those of other treatment groups (2 min; 93.5, 35.4%, 2.5 min; 76.9, 30.7%, 3 min; 88.8, 36.1% and 3.5 min; 83.7, 8.1%). 2) When the in vitro survival of vitrified groups according to different developmental stage was examined at 48 h after thawing using 1 min dilution method, the hatching rates of fast developed embryos (expanded blastocyst: 81.3%: early hatching blastocyst: 86.2%) were higher than that of delayed developed one (early blastocyst: 46.6%). 3) In addition, when the in vitro survival of vitrified groups according to different embryo age was compared, the hatched rates at 48 h after thawing of Day 7 (66.6%) and Day 8 embryos (60.0%) were significantly higher than that of Day 9 embryos (22.7%) (P＜0.05). These results demonstrate that vitrified bovine IVM/IVF/IVC blastocysts can be successfully survived in vitro using one-step dilution (1 min) method.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.17
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• pp.1-44
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• 1974

These studies were made at Suwon in 1972 and at Suwon, Iri, and Kwangju in 1973 to investigate grain filling process and variation of grain quality of NB 68513 and Caprock as hard red winter wheat, Suke ＃169 as soft red winter wheat variety and Yungkwang as semi-hard winter variety, grown under-three different fertilizer levels and seeding dates. Other experiments were conducted to find the effects of temperature, humidity and light intensity on the grain filling process and grain quality of Yungkwang and NB 68513 wheat varieties. These, experiments were conducted at Suwon in 1973 and 1974. 1. Grain filling process of wheat cultivars: 1) The frequency distribution of a grain weight shows that wider distribution of grain weight was associated with large grain groups rather than small grain group. In the large grain groups, the frequency was mostly concentrated near mean value, while the frequency was dispersed over the values in the small grain group. 2) The grain weight was more affected by the grain thickness and width than by grain length. 3) The grain weight during the ripening period was rapidly increased from 14 days after flowering to 35 days in Yungkwang and from 14 days after flowering to 28 days in NB 68513. The large grain group, Yungkwang was rather slowly increased and took a longer period in increase of endosperm ratio of grain than the small grain group, NB 68513. 4) In general, the 1, 000 grain weight was reduced under high temperature, low humidity, while it was increased under low temperature and high humidity condition, and under high temperature and humidity condition. The effect of shading on grain weight was greater in high temperature than in low temperature condition and no definite tendency was found in high humidity condition. 5) The effects of temperature, humidity and shading on 1, 000 grain weight were greater in large-grain group, Yungkwang than in small grain group, NB 68513. Highly significant positive correlation was found between 1, 000 grain weight and days to ripening. 6) The 1, 000 grain weight and test weight were increased more or less as the fertilizer levels applied were increased. However, the rate of increasing 1, 000 grain weight was low when fertilizer levels were increased from standard to double. The 1, 000 grain weight was high when planted early. Such tendency was greater in Suwon than in Kwangju or Iri area. 2. Milling quality: 7) The milling rate in a same group of varieties was higher under the condition of low temperature, high humidity and early maturing culture which were responsible for increasing 1, 000 grain weight. No definite relations were found along with locations. 8) In the varieties tested, the higher milling rate was found in large grain variety, Yungkwang, and the lowest milling rate was obtained from Suke ＃ 169, the small grain variety. But the small grained hard wheat variety such as Caprock and NB 68513 showed higher milling rate compared with the soft wheat variety, Suke ＃ 169. 9) There were no great differences of ash content due to location, fertilizer level and seeding date while remarkable differences due to variety were found. The ash content was high in the hard wheat varieties such as NB 68513, Caprock and low in soft wheat varieties such as Yungkwang and Suke ＃ 169. 3. Protein content: 10) The protein content was increased under the condition of high temperature, low humidity and shading, which were responsible for reduction of 1, 000 grain weight. The varietal differences of protein content due to high temperature, low humidity and shading conditions were greater in Yungkwang than in NB 68513. 11) The high content of protein in grain within one to two weeks after flowering might be due to the high ratio of pericarp and embryo to endosperm. As grains ripen, the effects of embryo and pericarp on protein content were decreased, reducing protein content. However, the protein content was getting increased from three or four weeks after flowering, and maximized at seven weeks after flowering. The protein content of grain at three to four weeks after flowering increased as the increase of 1, 000 grain weight. But the protein content of matured grain appeared to be affected by daily temperature on calender rather than by duration of ripening period. 12) Highly significant positive correlation value was found between the grain protein content and flour protein content. 13) The protein content was increased under the high level of fertilizers and late seeding. The local differences of protein content were greater in Suwon than in Kwangju and Iri. 14) Protein content in the varieties tested were high in Yungkwang, NB 68513 and Caprock, and low in Suke ＃ 169. However, variation in protein content due to the cultural methods was low in Suke ＃ 169. 15) Protein yield per unit area was increased in accordance with increase of fertilizer levels and early maturing culture. However, nitrogen fertilizer was utilized rather effectively in early maturing culture and Yungkwang was the highest in protein yield per unit area. 4. Physio-chemical properties of wheat flour: 16) Sedimentation value was higher under the conditions of high temperature, low humidity and high levels of fertilizers than under the conditions of low temperature, high moisture and low levels of fertilizers. Such differences of sedimentation values were more apparent in NB 68513 and Caprock than Yungkwang and Suke ＃ 169. The local difference of sedimentation value was greater in Suwon than in Kwangju and Iri. Even though the sedimentation value was highly correlated with protein content of grain, the high humidity was considered one of the factors affecting sedimentation value. 17) Changes of Pelshenke values due to the differences of cultural practices and locations were generally coincident with sedimentation values. 18) The mixing time required for mixogram was four to six minutes in NB 68513, five to seven minutes in Cap rock. The great variation of mixing time for Yungkwang and Suke ＃ 169 due to location and planting conditions was found. The mixing height and area were high in hard wheat than in soft wheat. Variation of protein content due to cultural methods were inconsistent. However, the pattern of mixogram were very much same regardless the treatments applied. With this regard, it could be concluded that the mixogram is a kind of method expressing the specific character of the variety. 19) Even though the milling property of NB 68513 and Caprock was deteriorated under either high temperature and low humidity of high fertilizer levels and late seeding conditions, baking quality was better due to improved physio-chemical properties of flour. In contrast, early maturing culture deteriorated physio-chemical properties, milling property of grain and grain protein yield per unit area was increased. However, it might be concluded that the hard wheat production of NB 68513 and Caprock for baking purpose could be done better in Suwon than in Iri or Kwangju area. 5. Interrelationships between the physio-chemical characters of wheat flour: 20) Physio-chemical properties of flour didn't have direct relationship with milling rate and ash content. Low grain weight produced high protein content and better physio-chemical flour properties. 21) In hard wheat varieties like NB 68513 and Caprock, protein content was significantly correlated with sedimentation value, Pelshenke value and mixing height. However, gluten strength and baking quality were improved by the increased protein content. In Yungkwang and Suk ＃ 169, protein content was correlated with sedimentation value, but no correlations were found with Pelshenke value and mixing height. Consequently, increase of protein content didn't improve the gluten strength in soft wheat. 22) The highly significant relationships between protein content and gluten strength and sedimentation . value, and between Pelshenke value, mixogram and gluten strength indicated that the determination of mixogram and Pelshenke value are useful for de terming soft and hard type of varieties. Determination of sedimentation value is considered useful method for quality evaluation of wheat grain under different cultural practices.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.85-92
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• 1999

This study was designed to determine effect of cryoprotectant kinds and cell stages on OPP vitrification method in mouse embryos. The freezing speed, cryoprotectants and cell stage could affect of embryo viability following various vitrification methods. The vitrification solution used were consisting of 40% (v/v) ethylene glycol, 18% (w/v) Ficoll, 0.3 M sucrose solution in holding medium (D-PBS supplemented with 5% FCS: HM) (EFS) or 16.5% ethylene glycol , 16.5% dimethyl sulfoxide, 0.5 M sucrose in HM (EDS). The embryos were collected from oviduct at 18 h after hCG injection and then washed and cultured in mHTF medium until use. In experiment 1, the blastocysts were vitrified by OPP straw to determine the optimal vitrification solution of EFS or EDS. The post-thaw survival rates at re-expanded stage rates were significantly different between EFS and EDS (95.0 vs 100%), but at hatching stage was not different between EFS and EDS (90.0 vs 95.0%). respectively. In experiment 2, zygotes, 2-, 4-cell, morula and blastocysts were vitrified by OPP method to determine the acceptable of early stage embryos. The development rates to expanded blastocyst in zygote (70.0%) were significantly lower rather than those in 2-, 4- 8-cell, compacted morula or blastocyst (89.7, 90.0, 92.8, 97.6 or 97.5%), respectively. However, the cell number of post-thaw developed to expanded blastocyst in blastocyst and control blastocyst stage (39.6$\pm$2.81, 35.7$\pm$2.98) were significanty higher than those in zygote, 2-, 4-, 8-cell, compacted morula (29.8$\pm$3.21, 31.3$\pm$3.83, 29.3$\pm$3.58, 28.9$\pm$3.21 or 30.8$\pm$2.93). In experiment 3, the zygotes were exposed in VSl for 1, 2, and 3 min to the optimal exposed time. The cleavage rates (91.6, 88.5, 88.9%) and develop mental rates to blastocyst (83.3, 74.3 and 69.4%) depends on the exposed time in VSl were not significantly different among 1, 2, or 3 min, respectively. The cell number also were not significantly different among exposed time in VS1. respectively. These results indicate that OPP method could be useful for vitrification either EFS or EDS vitrification solution. The post-thaw survival rates at zygote were significantly lower than those at 2-, 4-, 8-cell, morula or blastocyst, respectively. The zygote stage were more sensitive rather than late stage embryos. The exposing time in VS1 for 1 min was better than that for 2 or 3 min, even it was not significantly different. The OPP vitrification method could be useful of mouse embryos either with EFS or EDS vitrification solution.