• Clinical and Experimental Reproductive Medicine
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• v.33 no.2
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• pp.85-95
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• 2006

Objective: To investigate whether polymorphisms of gene encoding CYP1B1 is associated with the risk of endometriosis in Korean women. Methods: We investigated 199 patients with histopathologically confirmed endometriosis rAFS stage III/IV and 183 control group women who were surgically proven to have no endometriosis. The genetic distribution of four different CYP1B1 polymorphisms at $G^{119}-T$, $G^{432}-C$, $T^{449}-C$, and $A^{453}-G$ were analyzed by polymerase chain reaction(PCR) and restriction fragment length polymorphism of PCR products. Results: We found no overall association between each individual CYP1B1 genotype and the risk of endometriosis. The odds ratio of genotype GG/GC+GG/TC+TT/AA compared to GG/CC/CC/AA(reference) was calculated as 2.06 with a 95% confidence interval of 1.003~4.216. Conclusion: This results suggest that CYP1B1 genetic polymorphism may be associated with development of endometriosis in Korean women.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.379-384
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• 1996

This study was undertaken to investigate the sister chromatid exchange (SCE) frequency and embryonic development after exposure to cryoprotectants and vitrification of mouse zygotes. Mouse IVF zygotes were cryopreserved by vitrification using vitrification solution, EFS40 (40% ethylene glycol, 30% Ficoll and 0.3 M sucrose in phosphate buffer saline containing 10% FBS). After mouse zygotes were exposed to EFS40 at $25^{\circ}C$ for 30 sec., they were immediately plunged into $LN_2$ or cultured for cryoprotectant toxicity test without freezing. The results obtained in these experiments were summarized as follows: After thawing, survival rates to the 2-cell stage of zygotes exposed to or vitrified in EFS 40 (98.5%, 95.2%) were not significantly difference compared with that of control (100%). However, the developmental rates upto blastocyst and hatching blastocyst in vitrified groups (66.7, 50.0%) were lower than those of control (93.9, 81.8%) or exposed group (94.0, 78.8%) (p<0.05). When the influence of vitrification and exposure to cryoprotectant on the in vitro development of mouse zygotes was assessed by the SCE frequency, the SCE frequency in exposed ($20.2{\pm}2.1$) to or vitrified embryos ($21.4{\pm}3.2$) was higher than that in control embryos ($16.8{\pm}1.5$). These results suggest that the frequency of SCE was increased after cryoprotectant exposure or Vitrification although developmental rates of zygotes upto blastocysts and /or hatching blastocysts were not afected by cryoprotectant.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.3
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• pp.319-326
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• 1996

The objective of this study was to investigate correlation between the morphology by microscopic assessments of surplus blastocysts produced in human IVF program and their cell number obtained by differential labelling method. For these experiments, 76 surplus human blastocysts were obtained from 36 patients on day 5 after IVF, the embryos were classified to early (ErB), early expanding (EEB), middle expanding (MEB), expanded blastocyst (EdB) according to their blastocoel expansion and zona thickness. When the ovum size and zona thickness of the classified blastocysts were measured using micrometer, although the embryos were produced in the same culture condition, there were significant variances in ovum size ($148.8 217.6{\mu}m$) and zona thickness ($1.2-14.4{\mu}m$). Total blastomere cell number counted after hoechst staining was increased by two to three fold during the transition period from ErB ($39.1{\pm}3.6$) to EdB ($(89.6{\pm}3.3)$) stage on day 5 after IVF. ICM ($11.9{\pm}1.8-22.2{\pm}4.3$) and TE ($24.5{\pm}3.6-70.0{\pm}7.7$) cell numbers using differential labelling were also showed the increased pattern according to the developmental level. Especially, EdB which showed poor ICM morphologically also indicated the low ICM cell number after differential labelling. This demonstrated that there is good correlation between the morphological assessment and the cell number. The count of ICM and TE nuclei using differential labelling can be used as an important criterion, if it is accompanied with morphological assessments, in selecting the better embryos for improving the pregnancy rates in human blastocyst transfer program.

• Korean Journal of Ichthyology
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• v.9 no.1
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• pp.99-107
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• 1997

The early life history and reproductive ecology of Siniperca scherzeri were studied to obtain fundamental information in aquaculture and reinforcement of natural population in Soyang Lake, Buk-myon, Chunchon-shi, Kangwon-do from June to October 1996. Symptric species with adult fishes (+1 ages) of Siniperca scherzeri were 11 species belonging to 6 families and 10 genera and those with Juveniles (2~3 months) were 5 species belonging to 4 families and 5 genera. The sex radio of this species were 1 (female, 85) : 1.24 (male, 105). Adult and juvenile of this species were predominantly piscivores. Bluegill, Lepomis macrochirus, Zacco platypus, common carp, Cyprinus carpio, unidentified fish and shrimp were important components of the food items. The spherical eggs were demersal and separative without a colorless transparent chorion and slightly yellowish yolk containing one large oil globule (0.5~0.7mm). The egg just after fertilization were measuring 1.72~2.05mm (n=30), and expanded to 2.27~2.58mm (n=30) in diameter after 30 min. Hatching occurred 130~155 hrs after fertilization at water temperature of $20{\sim}25^{\circ}C$ and newly hatched larvae measuring 5.5~7.1mm in total length. In the newly hatched larvae, numerous branched malanophores were distributed on the yolk and abdomen of caudal peduncle. In ten-day old larvae, the yolk was mostly absorbed and the head spines and the teeth were well developed. All fin rays were formed and total length of the larvae were reached 13.6~15.6mm at 20 days after hatching. In fifty-five day old larvae were similar in both body shape and color to adult. The juvenile stage at 4 months after hatching were attained 86.4~95.3mm (n=7) in total length and 8.77~14.78g (n=7) in body weight.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.87-92
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• 1998

This study was carried out to investigate in vitro/in vivo development of vitrified mouse oocytes. Mouse oocytes were vitrified using EFS30, 35 and 40 (30, 35 and 40% ethylene glycol, 18% ficoll and 0.5 M sucrose in M2 medium). After being exposed or vitrified-thawed, oocytes of normal morphology were inseminated in vitro by $1-2\times10^6/ml$ of epididymal sperm. The rates of fertilization, in vitro/in vivo development and cell number (inner cell mass and tropectoderm cell) of blastocysts in each treatment group were examined. The results obtained in these experiments were summarized as follows: The cleavage rates were obtained in EFS35 containing 35% ethylene glycol higher than in EFS30 and EFS40. The development rate of vitrified-thawed oocytes to two-cell stage after in vitro fertilization (51.1%) was significantly different compared to that of exposed to vitrification solution without cooling (60.0%) and control (68.2%) (p<0.05). However, there were no differences in the blastocyst formation from the cleaved embryos among groups (75.0, 73.3 and 80.0%). Also, the mean number of cells per blastocysts of vitrified group $(92.5{\pm}2.9)$ was similar to that of the exposed $(98.5{\pm}5.3)$ and control $(100.9{\pm}4.8)$. In vivo development of the blastocysts derived from vitrified-thawed oocytes resulted in fetal development (50.7%) and implantation rates (80.0%) which are very similar to those of control (58.2, 78.2%). These results suggest that mouse oocytes could be cryopreserved using vitrification solution (EFS35) based on ethylene glycol.

• Korean Journal of Weed Science
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• v.7 no.1
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• pp.35-44
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• 1987

Differing in water conditions, the dry matter weight per plant was highest at 0 cm flooding depth, and was decreased at above 2 cm flooding depths. The shoot and spikes per pot developed best at flooding depths of 0 and 2 cm, but worst at -5cm or above 4cm flooding depths. The dry matter weight of shoots was linearly increased, but the weight of roots was sharply decreased according to high temperature after flower initation. The shoots and spikes per pot developed more effectively at 25$^{\circ}C$ than at 35 or 15$^{\circ}C$. The number of shoot and spike per pot were decreased according to higher shading. The effects of shading of 25-45% were not significantly small. The natural white and yellow spectra were the most effective to increase dry matter weight, shoots and spikes per pot. The dry matter weights of shoot and root per plant were not significantly different among at: 50 and 75% clay. The number of shoots were best and continuously increased at 75% clay, and the increments at 0 and 100% clays showed the lag period at early stage. The pattern of spikes was similar to shoots with less difference in various clay composition. The dry matter weight per plant in paddy field and upland field was basically similar. The numbers of shoot and spike were increased in response to increased fertilizer levels up to 20Kg per 10a of each component.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.3
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• pp.187-197
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• 2009

Objective: Phosphorylation and dephosphorylation of proteins are important in regulating cellular signaling pathways. Bead-based multiplex phosphorylation assay was conducted to detect the phosphorylation of seven proteins to maximize the information obtained from a single lysate of stage-specific mouse oocytes at a time. Methods: Cumulus-oocyte complexes (COCs) were cultured for 2 h, 8 h, and 16 h, respectively to address phosphorylation status of seven target proteins during oocyte maturation process. We analyzed the changes in phosphorylation at germinal vesicle (GV, 0 h), germinal vesicle breakdown (GVBD, 2 h), metaphase I (MI, 8 h), and metaphase II (MII, 16 h in vitro or in vivo) mouse oocytes by using Bio-Plex phosphoprotein assay system. We chose seven target proteins, namely, three mitogen-activated protein kinases (MAPKs), ERK1/2, JNK, and p38 MAPK, and other 4 well known signaling molecules, Akt, GSK-$3{\alpha}/{\beta}$, $I{\kappa}B{\alpha}$, and STAT3 to measure their phosphorylation status. Western blot analysis and kinase inhibitor treatment for ERK1/2, JNK, and Akt during in vitro maturation of oocytes were conducted for the confirmation. Results: Phosphorylation of ERK1/2, JNK, p38 MAPK and STAT3 was increased over 3 folds up to 20 folds, while phosphorylation of the other three signal molecules, Akt, GSK-$3{\alpha}/{\beta}$, and $I{\kapa}B{\alpha}$ was less than 3 folds. All of these results except for Akt were statistically significant (p<0.05). Conclusion: This is the first report on the new and valuable method measuring many phosphoproteins simultaneously in one minute sample such as oocyte lysates. All of the three MAPKs, ERK1/2, JNK, and p38 MAPK are involved in the process of mouse oocyte maturation. In addition, STAT3 might be important regulator of oocyte maturation, while Akt phosphorylation at Serine 473 may not be involved in the regulation of oocyte maturation.

• Reproductive and Developmental Biology
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• v.36 no.4
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• pp.261-267
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• 2012

The technique of SCNT is now well established but still remains inefficient. The in vitro development of SCNT embryos is dependent upon numerous factors including the recipient cytoplast and karyoplast. Above all, the metaphase of the second meiotic division (MII) oocytes have typically become the recipient of choice. Generally high level of MPF present in MII oocytes induces the transferred nucleus to enter mitotic division precociously and causes NEBD and PCC, which may be the critical role for nuclear reprogramming. In the present study we investigated the in vitro development and pregnancy of White-Hanwoo SCNT embryos treated with caffeine (a protein kinase phosphatase inhibitor). As results, the treatment of 10 mM caffeine for 6 h significantly increased MPF activity in bovine oocytes but does not affect the developmental competence to the blastocyst stage in bovine SCNT embryos. However, a significant increase in the mean cell number of blastocysts and the frequency of pregnant on 150 days of White-Hanwoo SCNT embryos produced using caffeine treated cytoplasts was observed. These results indicated that the recipient cytoplast treated with caffeine for a short period prior to reconstruction of SCNT embryos is able to increase the frequency of pregnancy in cow.

• Korean Journal of Animal Reproduction
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• v.16 no.1
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• pp.63-76
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• 1992

This study was carried out to investigate the appearance time of the second polar body for producing Gynogenesis or Triploid which could be obtained by arresting the second polar body by cold shock, and then blastoderm was used to measure fertility that revealed the nature of oogenesis, the effects of water temperature on fertility, hatchability, abnormality, viability and growth rate, and the water temperature and the breeding methods to prevent early death of larvae in Korean loach(Misgurnus anguillicaudatus) ; the results obtained in this study were summarized as follows. The second polar body was observed ont he surface of plasma disc close to micropyle within 10~40 min after fertilization at 29$^{\circ}C$. Artificial inseminatin had to be done immediately after the egg spawning because the spermatozoa of loach their mobility within 2 minutes when they were exposed to water. The amount of time needed to reach at blastoderm stage was 12 hours if fertilized eggs were incubated at 16$^{\circ}C$, 8 hours at 19$^{\circ}C$, 6 hours at 21$^{\circ}C$, 5 hours at 23$^{\circ}C$, 4 hours at 26$^{\circ}C$ and 3 hours 30 min at 29$^{\circ}C$ showing the shorter time for development of eggs at higher temperature. Fertilization rates in water temperatures of 19$^{\circ}C$, 21$^{\circ}C$, 23$^{\circ}C$, and 26$^{\circ}C$ were higher than those of water temperatures, 16$^{\circ}C$ and 29$^{\circ}C$. Water temperatures at 19$^{\circ}C$, 21$^{\circ}C$, and 23$^{\circ}C$ showed higher hatching rates that those of 16$^{\circ}C$, 26$^{\circ}C$, and 29$^{\circ}C$, while abnormal rates in 16$^{\circ}C$, 19$^{\circ}C$, 21$^{\circ}C$ and 23$^{\circ}C$ were lower than that of 26$^{\circ}C$ and 29$^{\circ}C$. Water temperatures at 16$^{\circ}C$, 19$^{\circ}C$, 21$^{\circ}C$, 23$^{\circ}C$ and 26$^{\circ}C$ respectively, were more different than 29$^{\circ}C$ in survival rates. The embryos were hatched at 72 hours after fertilization in 16$^{\circ}C$ water temperature, 48 hours in 19$^{\circ}C$, 40 hours in 21$^{\circ}C$, 32 hours in 23$^{\circ}C$, 25 hours in 26$^{\circ}C$, and 16 hours in 29$^{\circ}C$. Within three days after hatched out, the larvage grew 3mm in total length, the yolk granules were entirely consumed and the head and the trunk became thicker. Within 45 days after hatched out, the larva grew 25mm at 29$^{\circ}C$, 21mm at 26$^{\circ}C$, 16mm at 23$^{\circ}C$, 15mm at 21$^{\circ}C$, 12mm at 16$^{\circ}C$ in a 30 litreglass aquarium.

• Reproductive and Developmental Biology
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• v.31 no.1
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• pp.43-48
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• 2007

This study was conducted to examine the characteristics of fresh and frozen semen, proliferating efficiency by AI, and the coat color of offsprings in Korean Native Striped Cattle (Bos namadicus Falconer, Chikso). Semen were collected from 6 heads of tiger-coated male Chikso. In vitro fertilization (IVF) was conducted with frozen-thawed semen and in vitro matured Korean native brown cattle (general Hanwoo) oocytes. Total 18 heads of Hanwoo and Chikso were inseminated using Chikso semen. Coat colors of total 40 offsprings produced by AI were evaluated. The characteristics of the fresh and frozen-thawed Chikso semen did not differ among individuals. In vitro fertilization rate of Chikso semen was not different from that of general Hanwoo semen. However, developmental rate to the blastocyst stage of IVF embryos was higher in Chikso semen (25.9%) than in general Hanwoo semen (p<0.05). There was no difference in conception rate after AI between Chikso and general Hanwoo. The coat colors of offsprings varied, only 42.5% (17/40 heads) of offsprings had tiger coat color. Futhermore, only 55% of offsprings produced from the tiger-coated recipients had tiger coat color. This result shows that proliferation of Chikso by AI is possible, but further research approaches may be needed to enhance the productivity of tiger-coated Chikso.