• 한국가축번식학회지
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• 제11권1호
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• pp.73-84
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• 1987

This experiment was carried out to investigate the effects of hormone addition(FSH, HCG, estrogen and progesterone) and composition (BSA and FCS) of mKRB on the in vitro maturation and fertilizability of follicular oocytes of the Korean native cattle. The ovaries were removed at a slaughterhouse, returned to laboratory in a thermostat (30-35$^{\circ}C$) within 4 hr, and collected by aspirating normal follicles which had diameters of 1 to 6 mm. The oocytes with cumulus cells were cultured for 8, 16, 24 and 30 hr in a modified KRB solution containing BSA or FCS and hormones. The in vitro matured oocytes in mKRB containing FCS, FSH and steroids were transferred in the rabbit uterus for examination of their in vivo fertilizability with bovine sperm preincubated 4 to 6 hr in the rabbit uterus. 1. The mean number of oocytes collected per cattle was 6.5 from 1-3mm follicles, 1.3 from 4-6mm follicles, and total was 7.7. 2. The meiotic division at 16hr-cuture in the oocytes from 1-3mm follicles was slightly stimulated by the addition of FSH in mKRB + BSA solution compared with the control. At 30hr-culture, their maturation rates(%Met II) were also increased by FSH of 1 $\mu\textrm{g}$/ml(38.4%) and 5$\mu\textrm{g}$/ml(35.7%) as compared with the control (21.4%). The maturation rate at 30hr-culture in the oocytes from 4-6mm follicles was 53.8% and 57.1% by the FSH addition of 1$\mu\textrm{g}$/ml and 5$\mu\textrm{g}$/ml, respectively. These rates were similar with the control(57.1%), but higher than those of oocytes from 1-3mm follicles. 3. The meiotic division at 16hr-culture in the oocytes from 1-3mm follicles was stimulated by the HCG addition of 1IU/ml and 5IU/ml. However, the maturation rate at 30hr-culture was greatly decreased by the HCG addtion (26.6% and 13.3%) compared with the control(53.3%) and these rates (30.8%) in the oocytes from 4-6mm follicles were also lower than that fo the control(58.3%). 4. Low maturation rate (37.5%) of the oocytes cultured in mKRB containing BSA and 5IU/ml HCG was increased (55.0%) when 15% FCS with HCG was added to mKRB instead of BSA. 5. When 16hr-cultured oocytes in mKRB containing BSA and gonadotropins (5$\mu\textrm{g}$/ml FSH and 5IU/ml HCG) were transferred in the medium without gonadotropins and recultured for 16hr, the maturation rate of HCG-treated oocytes was greatly improved. 6. The maturation rates of oocytes were greatly affected by steroids. The combined addition of FCS+FSH+estrogen or +progesterone to mKRB increased the maturation rate compared with the combination of BSA+FSH or FCS+FSH in mKRB. 7. The fertilization rate, presence of pronuclei, was increased by the combination of FCS+FSH+p in mKRB as compared with that (5.6%) of BSA+FSH and the rates of FCS+FSH+steroids ranged from 12.5 to 17.6%.

• Reproductive and Developmental Biology
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• 제38권4호
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• pp.159-163
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• 2014

The objective of this study was to determine the effects of E. coli on boar sperm quality and reproductive performance in sows after artificial insemination. Three different levels of E. coli were artificially inoculated to semen with following concentrations; Control, 500, 5,000 and 50,000 colony forming unit (cfu)/ml. Semen samples were preserved at $17^{\circ}C$ for 5 days. Sperm motility was significantly decreased (p<0.05) on day 3 in the group inoculated with 5,000 cfu/ml compared to control groups. In all treatment groups, sperm motility was gradually decreased as storage time increased, but the decline pattern was more drastic in the groups inoculated with 5,000 and 50,000 cfu/ml groups from day 3 (p<0.05) compared to control group. After 3 day of storage at $17^{\circ}C$, sperm viability in sample inoculated with the highest concentration (50,000 cfu/ml) of bacteria was less (p<0.05) than that of control group. The pH of semen sample pH was maintained 7.2~7.5 in all groups during the experimental period. No differences (p>0.05) were found for both storage time and bacterial concentration. The pregnancy rate and live born piglets tend to decrease by increasing the concentration of E. coli in semen. In particular, the rate of pregnancy was lower in the group inoculated with 50,000 cfu/ml (58.3%) compare to the other groups (81.8, 75.0, 76.5%). These results suggest that the contamination of E. coli in boar semen negatively affects fertilizing ability of boar sperm and the reproductive performance obtained from sows after artificial insemination.

• Applied Microscopy
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• 제34권1호
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• pp.43-50
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• 2004

환경호르몬 비스페놀 A가 Xiphophorus maculatus의 생식에 미치는 영향을 알아보기 위하여 수컷을 100 ppb, 200 ppb, 400 ppb, 800 ppb의 농도에 3개월간 노출시킨 후 정상의 암컷과 교배시켜 치어수를 계수하여 수컷의 수정능력을 확인하였고, 치어를 6개월 동안 비스페놀 A에 노출시켜 치어의 암수비율, 기형율, 사망률 및 형태학적으로 정자형성과정을 조사하였다. 3개월간 비스페놀 A에 노출시킨 수컷과 정상인 암컷을 교배시켜 얻은 치어수는 정상대조군에서 평균 $19.00{\pm}3.02$마리였으나 비스페놀 A의 농도가 증가함에 따라서 치어수는 100 ppb에서 $11.50{\pm}2.22$마리, 200 ppb에서 $6.40{\pm}2.76$마리, 400 ppb에서 $5.70{\pm}1.34$마리 및 800 ppb에서 $3.70{\pm}2.11$마리로 감소하는 경향을 보였다. 치어의 암수비율은 비스페놀 A의 농도와 전혀 상관이 없었고 기형과 사망률은 0%였다. 노출시킨 지 6개월 후 정자형성과정은 800 ppb에서도 정상적으로 일어나고 있었다. 이상과 같이 비스페놀 A는 성전환, 수컷의 비율감소, 기형 및 사망에는 전혀 영향을 주지 않았고 단지 출생 개체수에만 영향을 주었다.

• Clinical and Experimental Reproductive Medicine
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• 제25권2호
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• pp.171-177
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• 1998

The combination of testicular sperm extraction (TESE) with ICSI can achieve normal fertilization and pregnancy rate and is effective method in obstructive and non-obstructive azoospermic patients. But, when pregnancy was not occurred, repeated testicular biopsies are not evitable. Therefore, in this study, we observed the survival rate of testicular spemratozoa and spermatozoa extracted from the seminiferous tubules after cryopreserved-thawed used for next IVF cycle with ICSI. In a total of 23 cases, obstructive azoospermia was 17 cases and non-obstructive azoospermia was 6 cases. In obstructive azoospermia, after thawing, motile spermatozua was observed in 13 cases (76.5%). The fertilization rate with 2PN was 67.6% and 5 pregnancies (29.4%) were achieved. In non-obstructive azoospermia, motile spermatozoa was observed in 2 case (33.3%) after thawing. The fertilization rates with 2PN was 53.7% and 3 pregnancies (50.0%) were achieved. A comparison of the results of motile spermatozoa after thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 3 cases (60.0%) and 12 cases (66.6%), respectively. The fertilization and pregnancy rates of thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section were 69.4% and 20.0%, 62.5% and 38.8%, respectively. Conclusively, thawed testicular spermatozoa and spermatozoa extracted from the thawed seminiferous tubule section can achieve normal fertilization and pregnancy and cryopreservation of testicular spermatozoa and seminiferous tubule may avoid repetition of testicular biopsies in azoospermic patients in whom the only source of spermatozoa is the testis.

• 한국수정란이식학회지
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• 제9권1호
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• pp.95-102
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• 1994

본 연구는 종모우의 선발방법으로 난포란을 이용하여 실험실내 정자의 수정능력을 직접 검정하여 평가코자 시도되었다. 즉 본 실험은 후대검정중에 있는 한우 후보종모우 15두의 동결융해정자의 수정능력을 평가하기 위하여 정액을 고장액(HIS)에 처리한 후 DM에서 6시간 그리고 소 난포액이 20% 첨가된 DM에서 4시간 전배양하여 수정능을 획득시켜 정자의 활력과 첨체 반응율을 조사하였고 전배양된 정자의 체내(토끼 난관) 또는 체외수정능력을 조사하기 위하여 FCS 15%, 발정암소혈정(CSS) 10%가 첨가된 mKRB에서 체외성숙된 한우난포란과 수정시켜 수정능력을 평가하였으며 인공수정에 의한 개체별 수태율과도 비교 검토한 바 다음과 같은 결론을 얻었다. 1. 한우 난포란의 체외성숙율은 BSA 첨가구 에서 43.8%, FCS 15% 첨가구에서 67.4%, CSS10% 첨가구에서 69.9%이었다. 2. 토끼 난관에서 체외수정율은 BSA 참가구에서 43.8%, FCS 15% 참가구 41.2% 및 CSS 10% 참가구 35.0% 이었다. 3. 후보종무우 15두의 정액을 HIS-DM으로 처리후 6시간 전배양하였을 때 정자의 활력지수는 9-32%였고 첨체반응율은 19-44% 이었으며 20% 난포액을 첨가하여 4시간 전배양 하였을 때 정자의 활력지수는 9-13% 이었고 첨체반응율은 20-43%로 개체간에 차이가 있었다. 4. 체외수정율은 6.6-85.7%였으며, 발정암소혈청(CSS) 10%가 첨가된 mKRB에서 성숙시킨 난포란이 FCS 15% 첨가된 mKRB에서 성숙시킨 난포란보다 다소 높았으나, 정자수정능획득방법간에는 차이가 없었 다. 5. 체외수정율에 있어서 전배양후 정자활력지수와는 부의 상관이 었으며, 첨체반응율과는 낮은 정의 상 관을 나타냈다. 6. 종모우의 수태율은 체외수정율, 정자활력지수 및 첨체반응율과 낮은 정의 상관관계를 나타냈다. 7. 종모우의 개체간 수태율 우열순위에서는 수정율순위와의 사이에 더욱 낮은 부의 상관관계를 보였다. 8. 이상의 연구결과 비록 후대정검중의 제한된 자료로 인하여 종모우 수태율과 체외수정율간에 유의적 인 상관관계는 없었으나, 연결 한우 수정율 평가에 대한 실험실내의 검정가능성을 찾을 수 있었다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1994년도 94 Symposium on Plant science September 10, 1994 Ewha Womans University, Seoul, Korea 94 식물학 심포지움 환경 스트레스와 식물의 반응
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• pp.29-39
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• 1994

Serious issues about the changes in the environmental conditions on earth associated with human activities have arisen, and the interest in these problems has increased. It is urgent to determine how the expansion of terrestrial UV-B radiation due to the stratospheric ozone depletion influences living matters. In this connection, we have been investigating the effects of UV-B radiation on the growth of rice cultivars (Oryza sativa L.). We report here some physiological and genetic aspects of resistance to inhibitory effects of UV-B radiation on growth of rice cultivars as described below. Elevated UV radiation containing large amount of UV-B and a small amount of UV-C inhibited the development of plant height, the photosynthetic rate and the chlorophyll content in rice plants in a phytotron. Similar results were obtained in experiments, in which elevated UV-V radiation. Similar results were obtained in experiments, in which elevated UV-B radiation (transmission down to 290 nm) was applied instead of UV-B radiation containing a small amount of UV-C. The inhibitory effects of UV radiation was alleviated by the elevated CO2 atmospheric environment or by the exposure to the high irradiance visible radiation. The latter suggested the possibility that the resistance to the effects of UV radiation was either due to a lower sensitivity to UV radiation or to a greater ability to recover from the injury caused by UV radiation through the exposure to visible radiation. The examination of cultivar differences in the resistance to UV radiation-caused injuries among 198 rice cultivars belonging to 5 Asian rice ecotypes (aus, aman, boro, bulu and tjeleh) from the Bengal region and Indonesia and to Japanese lowland and upland rice groups showed the following: Various cultivars having different sensitivities to the effects of UV radiation were involved in the same ecotype and the same group, and that the Japanese lowland rice group and the boro ecotype were more resistant. Among Japanese lowland rice cultivars, Sasanishiki (one of the leading varieties in Japan) exhibited more resistance to UV rakiation, while Norin 1 showed less resistance, although these two cultivars are closely related. It was thus indicated that the resistance to the inhibitory effects of UV radiation of rice cultivars is not simply due to the difference in the geographical situation where rice cultuvars are cultivated. Form the genetic analysis of resistance to the inhibitory effects of UV radiation on growth of rice using F2 plants generated by reciprocally crossing Sasanishiki and Norin 1 and F3 lines generated by self-fertilizing F2 plants, it was evident that the resistance to the inhibitory of elebated UV radiation in these rice plants was controlled by recessive polygenes.

• 한국가축번식학회지
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• 제17권4호
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• pp.311-323
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• 1994

This study was conducted to observe seasonal and individual changes in semen characteristics and sperm freezability, and sperm penetration into zona-free hamster eggs in Korean native goats. Buck response and change in semen characteristics to electrical stimulations was evaluated for four seasons throughout 2 years and percentage of motile sperm and normal apical ridge acrosome was investigated after equilibration and thawing for 4 seasons with 5 bucks. Sperm penetration rate was evaluated for 4 bucks. 1. Probe insertion at depth of 7cm and repeated stimulation for 3 sec was more effective(P<0.05) in buck response and semen collection than those of other conditions. 2. Semen characteristics from electrojaculation was signficantly(P<0.005) higher in spring and fall for semen volume, in spring and summer for sperm concentration and in fall for sperm motility than those in other seasons, respectively. However, there were no differences in total sperm among seasons. 3. Buck response to electrical stimulation showed significant difference(P<0.05) among individuals in all 3 seasons except winter. Significant individual difference in semen volume was only in spring and summer, but there was no indivudual difference in sperm concentration and total sperm in all season. 4. Washing of semen before freezing treatment was greatly(P<0.05) beneficial to sperm motility after thawing, no matter whether ejaculates exhibit egg yolk coagulation or not. 5. Sperm motility after glycerol equilibration was significantly(P<0.05) low in summer semen and motility after thawing was greatly(P<0.05) higher in winter semen than in other seasons. Freezability of unwashed sperm was significantly difference among bucks, but a yearly freezability of washed sperm after chilling and thawing were no differences among bucks and percentage of normal apical ridge acrosome were not different among seasons and bucks. 6. There was no significant difference in sperm motility after thawing between egg yolk levels in summer, although 20% level gave more higher motility than 5% level. 7. In summer, 3.2% glycerol and 3-h equilibration gave greatest percentage(P<0.05) of sperm motility and normal apical ridge acrosome after thawing. 8. Sperm penetration rate into zona-free hamster eggs was not different between bucks and seasons. Overall, it is concluded that to obtain maximum sperm output and successive semen freezing by electrojaculation method, buck selection with good response in all season could be basically considered and that seasonal effect on sperm freezability was more greater than that of individual bucks.

• 한국수정란이식학회지
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• 제26권1호
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• pp.65-70
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• 2011

희소 한우인 칡소의 정액 동결을 위해서 레시딘을 기본 희석제로 하는 AndroMed와 Tris-egg yolk extender를 사용하여 정자의 생존율과 활력 조사를 위해서 본 연구를 수행하였다. AndroMed 희석제를 사용하였을 때 생존율과 활력은 $73.4{\pm}11.2%$와 $67.9{\pm}14.6%$의 결과를 보였다. 그리고 Tris-egg yolk extender의 경우는 각각 $89.7{\pm}19.8%$와 $78.4{\pm}18.7%$ 결과를 보여 생존율에서는 Tris-egg yolk 희석제가 AndroMed 희석제를 사용하였을 때보다 유의적으로(p<0.05) 높은 결과를 보였다. 그러나 활력에서는 유의적인 차이를 보이지 않았으나 Tris-egg yolk 희석제에서 높은 경향을 보였다. 정액의 동결을 위해서 평형 시간에 따른 정자의 활력 조사를 위해서 육안적 방법과 CASA 프로그램 방법으로 조사를 실시한 결과, 처음 2시간부터 20시간까지 평형을 유도한 결과 육안적 방법보다는 CASA 프로그램 분석 방법의 결과 높은 수치를 확인할 수 있었다. AndroMed와 Tris-egg yolk extender 두 종류의 희석제로 사용된 동결 정액 사용으로 체외수정란을 생산 결과로서 분할율은 유의적인 차이를 보이지 않았으나(81.7% vs. 82.2), 배반포 발달율에서는 29.6% vs. 42.3%로서 Tris-egg yolk 희석제를 사용하였을 때 유의적으로(p<0.05) 높은 발달율을 보였다. 이러한 결과를 볼 때 희소 한우 품종인 침소 정액 동결을 위해서 Tris-egg yolk 희석제 사용이 동결된 정자의 생존율과 수정 능력 개선으로 실제적인 인공수정과 체외수정란 생산 및 가축유전자원 보존을 위해서도 효과적일 것으로 사료된다.

• 한국가축번식학회지
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• 제25권3호
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• pp.259-267
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• 2001

본 연구는 저정자증 또는 불임축의 수태율 증진과 불임해결에 적용할 목적으로, 일차적으로 소 정자의 PVP, HA의 농도, 신선 및 동결정자, 정자의 농도, 활력, 정자의 수정능획득법 및 난자의 투명대 부착과 미부착별로 현미조작에 의해 난자의 세포질내 단일정자의 주입했을 때 수정율 및 체외발생율을 조사하였다. 1. 소 난자의 ICSI시 PVP 농도를 0.01, 0.02, 0.03, 0.05%별로 처리한 정자를 micropipette내에 흡입하여 세포질내 주입하였을 때 수정율과 분할율은 각각 72.7∼90.9% 및 38.5∼54.5%로서 0.02%농도에서 비교적 높은 수정율과 분할율을 나타냈다. 2. 소 난자의 ICSI시 HA농도를 0.01, 0.02%, 0.02%의 PVP + HA별로 micropipette내에 흡입하여 세포질내 주입하였을 때 수정율과 분할율은 각각 72.7%와 81.8% 및 45.5%와 54.5% 및 83.3%와 37.5%였다. 3. 소 난자에 신선 및 동결정자를 이용하여 ICSI법으로 수정시 수정율은 각각 93.3%, 86.1%였으며, 분할율은 60.0%, 46.7%로서 신선정자를 이용하였을 때가 동결정자를 이용했을 때보다 높은 수정율과 분할율을 나타냈다. 4. Heparin, BFF 및 His법으로 수정능획득 처리한 정자로 IVF 및 ICSI시 수정율은 각각 61.9%, 52.6%, 45.0% 및 85.7%, 78.9%, 65.0%였으며, 분할율은 각각 23.8%, 15.8%, 10.0% 및 61.9%, 52.6%, 50.0%로서, 수정능획득 처리에 있어서 heparin법이 다른 수정능획득 처리법에 높은 수정율과 분할율을 나타났다. 5. 투명대의 부착과 미부착별 난자로 IVF 및 ICSI 법으로 수정시 수정율은 각각 63.2%, 47.8%와 84.2%, 78.3%였으며 분할율은 각각 15.8%, 8.7%와 57.9%, 34.8%였다. 6. 소 체외성숙 난자에 IVF법과 ICSI법으로 수정시켰을 때 수정율은 각각 63.3%, 64.6%와 88.2∼90.0%였으며, 분할율은 각각 26.7%, 29.2%와 52.9%, 67.5%로서 ICSI법으로 수정시켰을 때 수정율과 분할율이 크게 향상되었다.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.139-143
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• 2007

본 연구는 체세포를 이용하여 생산된 복제 한우 수소의 번식능력을 검토하기 위해 실시하였다. 복제 한우 수소(C-38 및 C-39) 또는 일반 한우 종모우로부터 정액을 채취하여 정자의 수 및 동결 전 후의 생존성 등을 살펴보았으며, 정자의 운동성 등은 computer assisted sperm analysis(CASA)를 이용하여 측정하였다. 또한, 이들의 수정 능력을 확인하기 위하여 체외수정과 인공수정을 각각 실시하였다. 정액 성상에서는 복제 수소들과 일반 종모우 간에 정액의 양, 정자의 농도 및 동결융해 후의 생존성 등에서 차이가 나타나지 않았다. CASA를 이용한 분석에서 운동성, 곡선 운동 속도(VCL), 직선운동 속도(VSL) 및 평균 진행 속도(VAP) 등은 복제 수소의 정액이 일반 종모우의 정액에 비하여 유의적으로 높았다 (p<0.05).체외수정에 따른 수정란의 분화율 및 배반포로의 발달율은 복제 수소와 일반종모우 간에 차이가 나타나지 않았다. 복제소 정액(C-38)을 이용하여 인공수정을 한 5두의 체세포 복제 대리모에서 암 수 각각 한 두씩의 건강한 복제 후대 송아지 2두를 생산하였다. 이상의 결과를 종합하여 보면, 실험에 공시된 복제 수소 개체간의 차이가 나타나기는 하였지만, 복제 수소는 정액 성상과 정자의 운동성 등에서 일반 종모우와 차이가 없었으며, 또한 인공수정을 통해 송아지를 생산함으로써 정상적인 번식능력이 있음을 확인하였다.