• Korean Journal of Animal Reproduction
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• v.23 no.4
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• pp.337-345
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• 1999

The present study was conducted to investigate the effects of various co-culture systems and supplemented protein sources on the in vitro development of bovine IVF embryos. Bovine cumulus oocyte complexes (COCs) were matured and fertilized in vitro. Presumptive zygotes with cumulus cells were transferred to TCM-199 or CRlaa containing 10% FBS or 3mg/$m\ell$ BSA, and cultured for 36~40 hr. After primary culture, cleaved embryos were co-cultured with cumulus cells(CC), bovine oviduct epithelial cells(BOEC) or Buffalo rat liver cells (BRLC) in TCM-199 or CRlaa supplemented with FBS or BSA respectively, for further 6 days. Cleavage rate increased with BSA(P＜0.01) in the both TCM-199(79%) or CRlaa(74%) When embryos were co-cultured with CC or BOEC in TCM-199, blastocyst development was enhanced with BSA(40% and 43%) compared to FBS (22% and 29%) , whereas in CRlaa no difference observed between BSA(40% and 39%) and FBS (40% and 42%). When embryos were co-cultured with BRLC monolayer, FBS enhanced the blastocyst development (P＜0.05) compared to BSA in both TCM-199(41% vs 31%) and CRlaa (44% vs 37%). The result of the present study showed that the cleavage rate of bovine IVF embryos increased with BSA, The result also showed that BSA can enhance the development of IVF embryos in co-culture with CC or BOEC in TCM-199, suggesting the in vitro development is affected by the medium and supplemented protein sources in co-culture with somatic cells.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.59 no.1
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• pp.73-82
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• 2014

This study was conducted to investigate the ear and kernel characteristics of waxy corn during ripening by double cropping, and to understand the pattern of starch accumulation in endosperm of waxy corn kernels. Chalok4 and Ilmichal were sown at April 20 (first cropping) and July 20 (second cropping) in 2011~2012. The accumulated temperature from silking to harvesting was about $590{\sim}630^{\circ}C$. It takes 23~24 days when Chalok4 and Ilmichal were sown in April 20, but July 20 sowing takes 32~35 days. Ear length and kernel set length were significantly shorter in second cropping (p<0.05). Kernel length, kernel width, 100-kernel weight, and starch content of waxy corn were increased as ears matured (p<0.05). Growth temperature was getting decreased during the ripening stage of second cropping, the rate of ear and kernel development had slowed. Starch granules started to accumulate in the cells around the pericarp, then developed in the cells around the embryo. In the second cropping, starch granules in the kernel of waxy corn were less compact than the first cropping, harvesting time of waxy corns can be extended. These results will be helpful to farmers for double cropping of waxy corn cultivation and management.

• Korean Journal of Ichthyology
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• v.26 no.1
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• pp.34-41
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• 2014

The present study describes the spawning ecology and early morphological development of Hemitripterus villosus. The natural spawning ground consisted of bedrock and pebbles was the intertidal coast at Taean (Chungnam) and its depth was about 5~10 m. Spawning period was mainly from the end of October to December, when the water temperature and salinity were $6.0{\sim}15.8^{\circ}C$ and mean 32.0‰, respectively. There were no difference of the body shape and color between female and male of Hemitripterus villosus, however its reproductive organs showed clear differences. The male had tube shaped genital papilla, which was connected with testis, and the female had seminal recepacle, which was the lower part of oviduct connected with ovary. Genital papilla of male came out of its body at spawning period and then male copulated. After copulation, female stored the sperm in its seminal recepacle and fertilized when it spawned. Fertilized eggs were reached 8 cells stage after fertilization at rearing water temperature $8.2{\sim}14.9^{\circ}C$. At 29 hours after fertilization, it reached morula stage, and at 146 hours after fertilization, its embryo was clearly formated. Hatching was begun from 1,488 hours (62 days) after fertilization with $8.2{\sim}14.9^{\circ}C$ water temperature. The newly hatched larvae were 12.99~15.46mm(mean $14.16{\pm}0.65$ mm) in TL (Total Length), and its mouth and anus were open. At 7 days after hatching, its yolk sac was completely absorbed and the myotomes were 15+25=40, measuring 15.23~15.54mm(mean $15.39{\pm}0.22$ mm, n=5) in TL. At 75~80 days after hatching, it was measured mean $30.06{\pm}0.76$ mm in TL, and it had reached the juvenile stage with the complete set of fin rays.

• Asian-Australasian Journal of Animal Sciences
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• v.28 no.10
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• pp.1394-1406
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• 2015

The AT motif-binding factor (ATBF1) not only interacts with protein inhibitor of activated signal transducer and activator of transcription 3 (STAT3) (PIAS3) to suppress STAT3 signaling regulating embryo early development and cell differentiation, but is required for early activation of the pituitary specific transcription factor 1 (Pit1) gene (also known as POU1F1) critically affecting mammalian growth and development. The goal of this study was to detect novel nucleotide variations and haplotypes structure of the ATBF1 gene, as well as to test their associations with growth-related traits in goats. Herein, a total of seven novel single nucleotide polymorphisms (SNPs) (SNP 1-7) within this gene were found in two well-known Chinese native goat breeds. Haplotypes structure analysis demonstrated that there were four haplotypes in Hainan black goat while seventeen haplotypes in Xinong Saanen dairy goat, and both breeds only shared one haplotype (hap1). Association testing revealed that the SNP2, SNP5, SNP6, and SNP7 loci were also found to significantly associate with growth-related traits in goats, respectively. Moreover, one diplotype in Xinong Saanen dairy goats significantly linked to growth related traits. These preliminary findings not only would extend the spectrum of genetic variations of the goat ATBF1 gene, but also would contribute to implementing marker-assisted selection in genetics and breeding in goats.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.1
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• pp.9-17
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• 2004

Objective: The aim of this study were to compare the effects of EG and PROH on cryopreservation of mouse and human embryos, and to find the optimal protocol for embryo freezing. Methods: Human embryos derived from fertilized eggs showing 3 pronuclei (PN) and mouse embryos were divided into two groups respectively: dehydrated with 1.5 M EG + 0.2 M sucrose or 1.5 M PROH + 0.2 M sucrose using the slow freezing method. Moreover mouse embryos were controlled the exposure time of cryoprotectant during dehydration or rehydration steps. Results: The survival rates of human embryos were 79.2% (84/106) in EG group and 77.9% (88/113) in PROH group. In mouse embryos, the survival and development rates up to blastocyst were 70.6% (245/347), 44.1% (123/279) in EG group and 62.1% (198/319), 45.1% (123/279) in PROH group, respectively. However, in EG group, partially damaged embryos after thawing were decreased compared to PROH group. In combination group, when the exposure time during dehydration and rehydration were reduced, the survival and embryonic developments were increased slightly, but not significant. Conclusion: Cryopreservation of mouse and human embryos at cleavage stage by using EG or PROH exhibited no statistical difference in the survival rate and/or developmental rate to blastocyst. However, the use of EG for cryopreservation of embryos might reduce the exposure time of the cryoprotectant because of a high permeation of EG and result in lessen its toxic effects.

• Development and Reproduction
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• v.2 no.2
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• pp.149-155
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• 1998

The polysialic acid (polySia) glycotope covalently modifies cell surface glycoconjugates on cells as evolutionarily diverse as microbes and human. The recent chemical identification of polysialylated glycoproteins in the jelly coat and on the cell surface of the sea urchin egg raises important questions about their biosynthesis and possible function. Using CMP-[$^{14}$ C]Neu5Ac as substrate and cell free preparations from eggs and embryos of the sea urchin Stronglylcentrotus nudus, we have identified a membrane associated CMP-Neu5Ac:poly-$\alpha$2, 8 sialosyl sialyltransferase (polyST) that transfers Neu5Ac to an endogenous acceptor. Optimal conditions for the polyST activity were found to be 2$0^{\circ}C$ in 20 mM MOPS buffer (pH 7.0). The polyST activity was increased 2.7 times by the addition of 10 mM $Mg^2$$^{+}$. The membrane-associated polyST also catalyzed the polysialylation of mammalian ganglioside GD3. Given that no structurally similar natural polysialylated gangliosides have been described, nor were observed in the present study, we conclude that a single polyST activity catalyzes sialylation of the endogenous acceptor and the gangliosides. Using an excess of GD3 as an exogenous acceptor, it was established that the expression of the polyST in S. nudus embryos increased rapidly at the mesenchyme blastula stage and reached at maximum at the gastrula stage. The finding that this polyST in the sea urchin embryo is developmentally regulated raises the possibility that it may play a role in the changing cell and tissue interactions that occur during gastrulation and the early stages of spicule formation.n.

• Development and Reproduction
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• v.10 no.1
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• pp.41-45
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• 2006

The spawning behavior of Liobagrus obesus was observed at Kumgang river, Yeongdong-gun, Chungcheongbuk-do from Apirl to July 2004. The fertilized eggs collected by dip net and skimming net were carried to the laboratory of Chonnam National University, and then egg, larvae and juvenils development were studied. Hatching of the embryo began about at 225 hrs 15 mins after morula stage in water temperature of $19.5{\sim}24.9^{\circ}C$(mean $22.8^{\circ}C$). The newly-hatched larvae were $7.30{\sim}7.90mm$(mean 7.66mm) in total length (TL), their mouth and anus were already opened with 14+28=42 myotomes. Sixteen days after hatching, the postlarvae were $13.00{\sim}14.05mm$(mean 13.48mm) TL, the yolk sac was completely absorbed. The juvenile stage was reached when all fin-rays were formed at 24 days after hatching, and $15.31{\sim}17.20mm$(mean 16.31mm) TL.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.63-69
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• 2001

This study was carried out to examine the effects of nitric oxide compounds (hemoglobin and L-NAME) on the development of porcine in vitro maturation (IVM) and in vitro fertilization (IVF) oocytes. Cumulus cell free embryos derived from porcine IVM/IVF oocytes were cultured in NCSU23 medium containing 1~5 $\mu\textrm{g}$/$m\ell$ hemoglobin added to 44 and 96hrs in culture times, and in NCSU23 medium containing 0, 10, 50 or 100mM L-NAME. The developmental rates beyond morulae stage in 0, 1 and 5 $\mu\textrm{g}$/$m\ell$ hemoglobin groups add to 44hrs in vitro culture times were 52.4%, 57.6% and 57.4%, respectively. The addition of hemoglobin groups made it slightly higher than the control group. The proportion of embryos developed to morulae and blastocysts in 1 $\mu\textrm{g}$/$m\ell$ hemoglobin add to 96hrs after in vitro culture (70.8%) was a little higher than those of 0 and 5 $\mu\textrm{g}$/$m\ell$ hemoglobin (66.2% and 62.8%). There was no significant difference in all groups (P〉0.05). The developmental rates beyond morulae stage in 0, 10, 50 and 100mM of L-NAME groups add to 96hrs after in vitro culture were 65.2%, 73.5%, 70.1% and 53.3%, respectively 10mM and 50mM L-NAME groups were significantly higher than in 0 and 100mM of L-NAME groups (P＜0.05). In conclusions, these results indicate that L-NAME (10mM, 50mM) can increase the proportion of embryos that develop into morulae and blastocysts but hemoglobin did not affect.

• Reproductive and Developmental Biology
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• v.34 no.3
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• pp.181-184
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• 2010

Osmolarity of culture media is one of the most important factors affecting in vitro development. This study was conducted to investigate the DNA methylation status of Pre-1 and satellite sequence in pig nuclear transfer (pNT) embryos produced under different osmolarity culture conditions. Control group of pNT embryos was cultured in PZM-3 for six days. Other two treatment groups of pNT embryos were cultured in modified PZM-3 with 138 mM NaG or 0.05M sucrose (mPZM-3, 320 mOsmol) for two days, and then cultured in PZM-3 (270 mOsmol) for four days. Previous our studies have reported that pNT embryos cultured in both hypertonic media showed significantly higher blastocyst formation rate than that of control. The DNA methylation status of the satellite sequences in blastocyst was characterized using bisulfite-sequencing technology. The satellite region had a similar methylation pattern of in vivo blastocyst among two culture groups excepting the control group. Each level of methylation is that the satellite DNA moderately methylated (43.10% of PZM-3; 56.12% of NaCl; 55.06% of sucrose; 60.00% of in vivo embryos). As a result of the sequence of PRE-1, CpG methylation pattern was similar to three groups, including in vivo group. In case of the satellite DNA region, the osmolarity conditions were affected CpG DNA methylation status while PRE-1 sequence was not affected CpG DNA methylation in pNT blastocyst stage. These results indicate that the modification of osmolarity in a culture media may influence to spatially change of DNA methylation of repetitive sequence for pNT embryo development.

• Development and Reproduction
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• v.8 no.1
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• pp.35-42
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• 2004

Oocyte maturation refers to the process that prophase I arrested germinal vesicle(GV) drives the progression of meiosis to metaphase II(MII) to have the capacity for fertilization and embryo development. To better understand the molecular mechanism(s) involved in oocyte maturation, we identified differentially expressed genes(DEGs) between GV and MII mouse oocytes using a new innovative annealing control primer (ACP) technology. Using 20 ACPs, we successfully cloned 32 DEGs between GV and Mll oocytes, and 26 out of these 32 DEGs were functionally known genes. Four genes including Pscd2 were GV-specific, 10 genes including PKD2 and CSN3 were highly expressed in GV oocytes(GV-selective), and 12 genes including Diva were highly expressed in MII oocytes (MII-selective). Ail of the genes identified in this study were first reported in the oocyte expression using ACP system and especially, we could characterize the existence of PKD-CSW signaling pathwayin the mouse oocytes. Results of the present study would provide insight for studying molecular mechanisms regulating oocyte maturation.