• Asian-Australasian Journal of Animal Sciences
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• 제21권3호
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• pp.358-363
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• 2008

The intracytoplasmic sperm injection (ICSI) procedure has recently been utilized to produce transgenic animals and may serve as an alternative to the conventional pronuclear microinjection in species such as pigs whose ooplasm is opaque and pronuclei are often invisible. In this study, the effects of sperm membrane disruption and electrical activation of oocytes on in vitro development and expression of transgene green fluorescent protein (GFP) in ICSI embryos were tested to refine this recently developed procedure. Prior to ICSI, sperm heads were treated with Triton X-100+NaCl or Triton X-100+NaCl+NaOH, to disrupt membrane to be permeable to exogenous DNA, and incubated with linearized pEGFP-N1 vector. To induce activation of oocytes, a single DC pulse of 1.3 kV/cm was applied to oocytes for $30{\mu}sec$. After ICSI was performed with the aid of a micromanipulator, in vitro development of embryos and GFP expression were monitored. The chemical treatment to disrupt sperm membrane did not affect the developmental competence of embryos. 40 to 60% of oocytes were cleaved after injection of sperm heads with disrupted membrane, whereas 48.6% (34/70) were cleaved without chemical treatment. Regardless of electrical stimulation to induce activation, oocytes were cleaved after ICSI, reflecting that, despite sperm membrane disruption, the perinuclear soluble sperm factor known to mediate oocyte activation remained intact. After development to the 4-cell stage, 11.8 (2/17, Triton X-100+NaCl+NaOH) to 58.8% (10/17, Triton X-100+NaCl) of embryos expressed GFP. The expression of GFP beyond the stage of embryonic genome activation (4-cell stage in the pig) indicates that the exogenous DNA might have been integrated into the porcine genome. When sperm heads were co-incubated with exogenous DNA following the treatment of Triton X-100+NaCl, GFP expression was observed in high percentage (58.8%) of embryos, suggesting that transgenic pigs may efficiently be produced using ICSI.

• 가정과삶의질연구
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• 제23권5호
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• pp.133-143
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• 2005

The purpose of this study was to gain understanding about residential satisfaction and preference of the members of the future elderly generation who have an intention to develop senior congregate housing. The data for this study were collected from 600 participants who were in their 40s to 60s, residing in Seoul, and who have an intention to develop senior congregate housing. The data collection spanned from December 1st to 20th, 2004. The results of this study were as follows: Firstly, the respondents who had a higher level of education, higher income, and a high-ranking job showed relatively higher residential satisfaction with their present house. Similar satisfaction level came between their present house and expecting ones in their later life. Financial responsibility was the most serious expected problem when they continue living in the present house in the later life. It means that the economic ability is the primary factor that determines whether they will have a stable life later. Secondly, as for the preference for the senior housing developer type, the group who preferred collectively developed senior congregate housing far outnumbered the other who preferred independent developer type. Also, the group who preferred collective developer type had higher economic ability than that of the other. These results indicate that above all, economic competence is the must significant variable, which should be considered in the future life and residential planning for the elderly. Korean senior congregate housing models should be developed based upon elaborate research. It is also necessary to develop different types of housing, such as a more practical type and a high quality type, as well as to respond to different kinds of developmental objectives, such as marketability and public benefit.

• 대한한방부인과학회지
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• 제21권1호
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• pp.257-266
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• 2008

Purpose: These experiments were undertaken to evaluate the effect of Onpoeum on ovarian functions and differential gene expressions related with cell viabilities caspase-3, MAPK and MPG in female mice. Methods: We administered the Onpoeum to 6-week-old female ICR mice for 4, 8, or 12 days. With different concentration of Onpoeum, the female mice were injected PMSG and hCG for ovarian hyperstimulation. The mice divided into 3 different groups for each experiment. We chose the Caspase-3 for cell apoptosis, MAPK and MPG genes for cell viability and DNA repair. Results: In case of 4, 8, 12 day of Onpoeum, we were examined the mean number of total ovulated oocytes and the number of morphologically normal oocytes. We were also examined the embryonic developmental competence in vitro. In addition we were examined the differential expression of cell apoptosis, viability and DNA repair related genes, Caspase-3, MAPK and MPG according to concentration and duration of Onpoeum. From these results showed that the administration of Onpoeum played a role of prevention of cell apoptosis and DNA damages and also increased cell proliferation resulted in ovarian functions. Conclusions: It is suggested that the medication of Onpoeum may have beneficial effect on reproductive functions of female mice via prevention of cell apoptosis and DNA damaging and promotion of cell proliferation.

• 미생물학회지
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• 제40권2호
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• pp.178-182
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• 2004

Saccharomyces cerevisiae의 class I 키틴생합성 효소인 Chsl의 활성측정법을 적용하여 Aspergillus nidulans의 영양균사에서 class I 키틴생합성 효소인 ChsC의 활성측정을 시도하였다. 그 결과, A. nidulans의 class I 키틴생합성효소도 효모류의 경우와 마찬가지로 트립신 처리에 의하여 활성화되는 효소전구체형태(zymogenic form)로 존재함을 알 수 있었다. 그리고 A. nidulans 야생형의 class I 키틴 생합성 효소활성은 트립신 처리에 의하여 6배 가량 증가되었다. 반면, chsC 유전자가 파괴된 돌연변이주는 트립신 처리에 의하여 효소활성이 증가되지 아니하였을 뿐만 아니라, 효소활성의 수준도 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성과 거의 동일한 수준이었다. 따라서, 트립신을 처리하여 측정한A. nidulans 야생형의 class I 키틴생합성 효소활성 값에서 트립신을 처리하지 아니한 야생형의 class I 키틴생합성 효소활성을 제외한 값이 A. nidulans 야생형의 ChsC 효소활성임을 알았다. 이러한 조건을 토대로 영양균사 생장과정 동안 ChsC의 효소활성을 측정한 결과, chsC 유전자의 발현양상과 유사하게 액체배양상태의 영양균사가 무성분화능을 획득하는 시기로 알려진 시간대에 효소의 활성이 증가하였다. 이러한 결과는 이미 보고된 바와 같이 chs 유전자가 A. nidulans의 영양균사 생장에 관여함을 시사하고 있다.

• Reproductive and Developmental Biology
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• 제36권3호
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• pp.141-145
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• 2012

The present study was conducted to develop a simple method for porcine oocyte maturation without $CO_2$ regulation. In experiment 1, we evaluated that the effect of $CO_2$ non-supplement on porcine oocyte maturation. Cumulusoocyte complexes (COCs) were collected from 2~6 mm follicles and divided into three groups (Control, tube-$CO_2$, and tube-non-$CO_2$). For control, COCs were cultured in 4-well multidish in a $CO_2$ incubator. For tube-$CO_2$, COCs were cultured in a round-bottom tube in a $CO_2$ incubator, and for tube-non-$CO_2$, COCs were cultured in a round-bottom tube sealed tightly without $CO_2$ supplement in a dry incubator. The proportion of oocytes reached to metaphase II (M-II) was not significantly different among three groups (87.9% to 91.4%). In experiment 2, we evaluated the effect of $CO_2$ non-supplement during oocyte maturation on development of embryos. Oocytes with a polar body were divided into two groups (Control and tube-non-$CO_2$) and applied 1.1 kV/cm or 1.2 kV/cm voltages for parthenogenetic activation. After activation, embryos were cultured for 6 days and examined the development. The proportion of embryos cleaved was not significantly different among treatment (86.3% to 91.5%). The proportion of embryo reached to blastocyst stage was not significantly different among treatment (13.9% to 25.2%). The cell number of blastocysts was not significantly different among treatment (29.0 to 32.4). In conclusion, oocytes cultured in a dry incubator without $CO_2$ supplement have enough competence to development after parthenogenetic activation. These results would be useful for transporting oocytes or embryos a long distance.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.718-724
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• 2007

Considerable attention has been focused on the cloning of mammalian embryos, as a consequence of poor development, in order to enhance the application of genetic engineering. Experiments were conducted to compare the developmental competence of parthenotes and reconstructed (NT) rabbit eggs with fetal fibroblasts (FFs) following various activation regimens. Oocytes and NT eggs were exposed to: electric stimulation (EST, Group 1) and EST followed by 6-dimethylaminopurine (DMAP, Group 2), cycloheximide (CHX, Group 3) or DMAP/CHX (Group 4). Pronuclear (PN) status, cleavage, blastocyst development and the ploidy were assessed. In parthenote groups 1, 2, 3 and 4, the PN formation differed significantly. And, the cleavage and blastocyst rates were 41.7 and 5%, 75.6 and 53.7%, 68 and 36%, 82.1 and 52.6%, respectively, among treatments. Polyploidy was observed in 17.2% of EST plus DMAP and 44.9% of EST plus DMAP/CHX groups. In SCNT groups (Group 1, 2, 3 and 4), the cleavage and blastocyst rates were 28.6 and 7.1%, 58.3 and 29.2%, 56.8 and 24.1%, 64.5 and 27.8%, respectively. The chromosomal composition differed significantly (p<0.05) among treatments. In Group 2 and 3, 53.8% and 81.8% of embryos revealed diploid chromosomal sets, respectively. However, in Group 4, 53.3% of embryos showed abnormal ploidy (mixoploid). Although DMAP or combination with DMAP/CHX resulted in higher in vitro development of rabbit SCNT embryos, higher incidence of chromosomal abnormality may induce problems related to fetal loss of at late stage of development.

• 대한수의학회지
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• 제57권2호
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• pp.89-95
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• 2017

This study was conducted to determine the effects of biophoton treatment during in vitro maturation (IVM) and/or in vitro culture (IVC) on oocyte maturation and embryonic development in pigs. An apparatus capable of generating homogeneous biophoton energy emissions was placed in an incubator. Initially, immature pig oocytes were matured in the biophoton-equipped incubator in medium 199 supplemented with cysteine, epidermal growth factor, insulin, and gonadotrophic hormones for 22 h, after which they were matured in hormone-free medium for an additional 22 hr. Next, IVM oocytes were induced for parthenogenesis (PA) or provided as cytoplasts for somatic cell nuclear transfer (SCNT). Treatment of oocytes with biophoton energy during IVM did not improve cumulus cell expansion, nuclear maturation, intraoocyte glutathione content, or mitochondrial distribution of oocytes. However, biophoton-treated oocytes showed higher (p < 0.05) blastocyst formation after PA than that in untreated oocytes (50.7% vs. 42.7%). In an additional experiment, SCNT embryos produced from biophoton-treated oocytes showed a greater (p < 0.05) number of cells in blastocysts (52.6 vs. 43.9) than that in untreated oocytes. Taken together, our results demonstrate that biophoton treatment during IVM improves developmental competence of PA- and SCNT-derived embryos.

• Clinical and Experimental Reproductive Medicine
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• 제41권3호
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• pp.125-131
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• 2014

Objective: Under estrogen deficiency, blastocysts cannot initiate implantation and enter dormancy. Dormant blastocysts live longer in utero than normal blastocysts, and autophagy has been suggested as a mechanism underlying the sustained survival of dormant blastocysts during delayed implantation. Autophagy is a cellular degradation pathway and a central component of the integrated stress response. Reactive oxygen species (ROS) are produced within cells during normal metabolism, but their levels increase dramatically under stressful conditions. We investigated whether heightened autophagy in dormant blastocysts is associated with the increased oxidative stress under the unfavorable condition of delayed implantation. Methods: To visualize ROS production, day 8 (short-term dormancy) and day 20 (long-term dormancy) dormant blastocysts were loaded with $1-{\mu}M$ 5-(and-6)-chloromethyl-2', 7'-dichlorodihydrofluorescein diacetate, acetyl ester (CM-$H_2DCFDA$). To block autophagic activation, 3-methyladenine (3-MA) and wortmannin were used in vivo and in vitro, respectively. Results: We observed that ROS production was not significantly affected by the status of dormancy; in other words, both dormant and activated blastocysts showed high levels of ROS. However, ROS production was higher in the dormant blastocysts of the long-term dormancy group than in those of the short-term group. The addition of wortmannin to dormant blastocysts in vitro and 3-MA injection in vivo significantly increased ROS production in the short-term dormant blastocysts. In the long-term dormant blastocysts, ROS levels were not significantly affected by the treatment of the autophagy inhibitor. Conclusion: During delayed implantation, heightened autophagy in dormant blastocysts may be operative as a potential mechanism to reduce oxidative stress. Further, ROS may be one of the potential causes of compromised developmental competence of long-term dormant blastocysts after implantation.

• Clinical and Experimental Reproductive Medicine
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• 제46권4호
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• pp.189-196
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• 2019

Objective: We aimed to evaluate the effects of different oxygen conditions (20% [high O₂], 5% [low O₂] and 5% decreased to 2% [dynamic O₂]) on mouse pre- and peri-implantation development using a novel double-channel gas supply (DCGS) incubator (CNC Biotech Inc.) to alter the oxygen concentration during in vitro culture. Methods: The high-O₂ and low-O₂ groups were cultured from the one-cell to the blastocyst stage under 20% and 5% oxygen concentrations, respectively. In the dynamic-O₂ group, mouse embryos were cultured from the one-cell to the morula stage under 5% O₂ for 3 days, followed by culture under 2% O₂ to the blastocyst stage. To evaluate peri-implantation development, the blastocysts from the three groups were individually transferred to a fibronectin-coated dish and cultured to the outgrowth stage in droplets. Results: The blastocyst formation rate was significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The total cell number was significantly higher in the dynamic-O₂ group than in the low-O₂ and high-O₂ groups. Additionally, the apoptotic index was significantly lower in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. The trophoblast outgrowth rate and spread area were significantly higher in the low-O₂ and dynamic-O₂ groups than in the high-O₂ group. Conclusion: Our results showed that a dynamic oxygen concentration (decreasing from 5% to 2%) had beneficial effects on mouse pre- and peri-implantation development. Optimized, dynamic changing of oxygen concentrations using the novel DCGS incubator could improve the developmental competence of in vitro cultured embryos in a human in vitro fertilization and embryo transfer program.

• 한국보육지원학회지
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• 제11권3호
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• pp.129-148
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• 2015

본 연구는 국내에서 이루어진 유아 수세기(counting) 관련 연구물의 연구동향을 알아보고자 수행되었다. 이를 위해 수세기를 주제어로 다루고 있거나 제목으로 명시한 국내의 학위 및 학술지 논문 59편을 선정하였고, 연구시기, 연구내용, 연구방법별 분석기준을 마련하여 분석을 실시하였다. 분석 결과 첫째, 시기별로 보면 수세기 연구물은 2001-2005년 사이에 현저하게 많았고 그 이외의 시기 동안에는 비슷한 비율로 발표되고 있는 것으로 나타났다. 둘째, 내용별로 보면 수세기의 발달적 경향을 알아보는 연구가 가장 많았고 다음은 수세기와 기타 수학적 개념 및 능력과의 관련성을 알아보는 연구가 많았다. 셋째, 연구방법별로 보면 양적 연구가 절대 다수를 차지하고 있었고 질적 연구는 매우 미미했으며, 연구대상은 3-5세의 일반 유아에게 집중되어 있었다. 본 연구결과를 통해 향후에는 이전에 확인된 내용들에 대해 중복적인 연구를 피하고 새로운 주제가 탐색될 필요가 있으며 영아를 대상으로 한 질적 연구가 활성화될 필요가 있음을 제안하였다.