체외 성숙시킨 M II기 난자의 여러 화학물질간의 중복 및 병용처리를 실시하여 수핵란으로서 적합한 활성화 조건과 활성화 후 자체 탈핵을 유도함으로써 보다 효율적인 탈핵 여건을 확립하고자 본 연구를 실시하였다. 1. Ethanol을 사용하여 6-DMAP 또는 cycloheximide 간의 단독, 중복, 그리고 병용처리를 실시한 결과, 활성화율, 난할율 및 체외 발달율에서 단독처리와 병용처리간의 차이는 없었으나 중복처리시 유의적으로 저하되었다(P<0.05). 2. Ca/sup 2+/ ionophore를 가지고 활성화 조건별로 처리하였을 경우, 6-DMAP와 병용 처리가 활성화율, 난할율 및 체외 발달율에서 각각 80.8%, 78.3%, 40.6%로 단독처리시의 58.0%, 62.9%, 27.0%보다 유의적으로 높았고(P<0.05), 또한 cycloheximide와의 병용처리도 활성화 및 난할율에서 단독처리와의 유의적 차이를 보였다(P<0.05). 그러나 중복처리시에는 효과를 나타내지 못하였다. 3. 탈핵전 수핵란에 미리 활성화 처리를 한 후 핵이식란을 재구성 한 결과 탈핵율 및 세포 융합율에서 90.7%, 71.8%로 활성화 처리하지 않은 것 77.8%, 61.1% 보다 유의적으로 높았다(P<0.05). 그러나 난할율 및 체외 발달율에서는 유의적으로 저하되었다(38.7%, 19.3% vs 68.8%, 30.6%, P<0.05). 또한 변성율도 활성화 처리하지 않은 구에 비해 유의적으로 높게 나타났다(P<0.05). 이상의 결과로 미루어 탈핵전 활성화하여 수핵란으로 이용할 경우 Ca/sup 2+/ ionophore와 6-DMAP를 병용 처리가 가장 효과적이며, 주입되는 체세포와 수핵란 간의 적정한 세포주기의 조절이 효율적인 돼지 복제수정란 생산을 위해 필요할 것이다.

활성화를 통한 수핵란의 대량확보를 위해 44시간동안 체외 성숙된 돼지 난자를 ethanol, Ca/sup 2+/-ionophore, 6-DMAP 및 cycloheximide의 화학물질들을 사용하여 단위발생을 유기한 후 그들의 가장적합한 처리농도 및 노출 시간을 규명하였다. 1. Ethanol은 10%, 10분 처리가 전핵형성율, 난할율 및 배발달율에 있어 각각 약 53.4%, 51.6%, 그리고 39.9%로 가장 적합한 조건으로 판명되었다. 2. Ca/sup 2+/-ionophore 가장 적합한 난활성화 조건은 25μM에서 2분간 처리한 것이며, 전핵형성율, 난할율 및 배발달율은 각각 약 59.7%, 62.2%, 그리고 43.9%를 보였다. 3. 6-DMAP를 처리하여 돼지 난자의 활성화를 유기하였을 경우 2mM의 농도에서 각각 약 57.3%, 58.4% 및 29.0%의 전핵형성율, 난할율, 그리고 배발달율을 보여 가장 적합한 조건을 보였으며 2시간∼4.5시간 사이의 노출에는 영향을 받지 않았다. 4. Cycloheximide는 5㎍/ml의 농도가 전핵형성율 2.1%, 난할율 47.7%, 배발달율 31.8%로 가장 은 효율을 보였고, 노출시간에서는 4시간∼6시간 동안 처리하였을 때 60.5∼65.8%, 63.6∼66.7% 및 39.0∼39.5%로 가장 적합한 조건으로 판명되었다. 이상의 결과들은 돼지 체외 성숙 난자의 활성화에 있어 각 화학물질들의 적합한 조건을 바탕으로한 중복처리 및 병용처리 조건 확립 및 효율적인 수핵란의 확보에 기여할 수 있을 것이다.

본 연구는 한우 체외수정란에 외래 유전자를 미세현미 주입한 후 체외 배발달을 조사하였다. DNA 미세주입은 체외수정 18~20시간 후에 DNA를 미세주입하였으며 체외 배발달율은 7일간 배양 후 조사하였다. 미세현미 주입 후 난할율은 36.3%로 대조구의 난할율 66.4% 보다 유의적으로 낮았으며(p<0.05) DNA가 주입된 수정란 중 상실배와 배반포배까지 발달율은 각각 5.6%와 1.9%로 대조구의 20.5%와 12.8%에 비하여 유의적으로 낮게 나타

This study was conducted to investigate the effect of hormones, protein sources and anti-oxidants on in vitro maturation (IVM) and in vitro fertilization(IVF) of bovine follicular oocytes. The rates of Holstein follicular oocytes classified as grade A and B(50.2% and 33.2%) were higher than those of Hanwoo cattle(40.3% and 32.0%, P<0.05). The cumulus cell expansion rates of oocytes cultured in TCM-199 and Ham's F-10 medium supplemented with 10% FCS and hormones were higher (81.9~87.6%) than those of non-treated groups (74.5~81.7%). The fertilization rates of oocytes cultured in TCM-199 and Ham's F-10 medim supplemented with 10% FCS, 1% BSA and 10% bFF was 53.8~55.0%, 51.4~52.6%, and 47.0~50.0%, respectively. The polyspermy rates was 13.6~14.2%, 10.0~11.1%, and 10.0%, respectively. When the oocytes were cultured in TCM-199 and Ham's F-10 medium with 50 -tocopherol, the fertilization rates was 62.0 and 60.2%, respectively. In the maturation medium added of 100 cysteamine, the fertilization rates was 64.7 and 66.7%, respectively. The fertilization and polyspermy rates of treated groups were higher than those of non-treated group. The results show that hormones, protein sources and anti-oxidants can provide a benefit for in vitro maturation and fertilization of bovine follicular oocytes.

The effect of several potential antioxidants were examined as a means of increasing the in vitro development of in vitro matured and in vitro fertilized oocytes into morulae and blastocysts. Korean native cattle embryos after in vitro fertilization were cultrued for 7 days at 38.5 in CR1aa containing varing concentration of the antioxidants in a gas phases consisting of 5% CO2, 95% humidified air. The results obtained were summarized as follows; The proportion of embryos developed to morulae and blastocysts in CR1aa containing 2.5uM -tocopherol(11.0% and 6.0%) was significantly higher than those of 0, 5.0, and 7.5uM -tocopherol (P<0.05). concentration of 50uM L-ascorbic acid (7.5% blastocysts) did affect the proportion of embryos developing into blastocystes(P>0.05). Addition of 200uM cysteamine was significantly higher than those of 0, 100 and 300uM (P<0.05). When the fertilized oocytes were cultured at 0. 200, 400 and 600uM of selenium for 168 hrs, the morulae rates were 12.2, 5.2, 16.0 and 16.1% respectively, and addition of 200uM selenium was significantly higher than those of 0, 400, 600uM (P<0.05). These results suggested that the addition of -tocopherol, L-ascorbic acid, cysteamine and selenicum can enhanced development to the morulae and blastocysts of in vitro derived fertilized oocytes.

This study was conducted to investigate the effect of -tocopherol and cysteamine with Whitten's medium in supporting the development on in vitro maturation(IVM), in vitro fertilization (IVF) and in culture(IVC) on porcine oocytes. When the immature oocytes were cultured of -tocopherol for 40h, the nuclear maturation rates were 39, 4, 52.5 and 54.1%, respectivley. The nuclear maturation rates of treat groups were signficantly (P<0.05) higher than those of non-treat groups. After matureation, the oocytes were inseminated in vitro in medium 199 with ejaculated spermatoza for examination of sperm penetration, polyspermy, male pronuclear(MPN) formation, and cleavage rate. Sperm penetration rates of treat higher than the control groups(P<0.05), and MPN formation rates were significantly(P<0.05) higher on treated groups (24.3~53.1%) than control groups(14.2~21.4%). After insemination, the cleavage rates at 120hr were groups higher than control groups(P<0.05).

The effects of acetic acid (AA) on aerobic plate counts (APC), gram-negative bacterial counts (GNC), and generation time (GT) in chicken wings stored at 4℃ were assessed. Chicken wings were treated with 0.5-1.5% (v/v) AA at exposure times of 5 min. Treatments of AA for 5 min significantly (P$lt;0.05) reduced aerobic plate counts (APC) and gram-negative bacterial counts (GNC) on the surface of chicken wings for 8 days, respectively. After 4 days of storage, treatments of 1.0% AA and 1.5% AA for 5 min completely (P$lt;0.05) inhibited APC and GNC compared to initial controls. Based on these results, treatments of 1.0% AA and 1.5 % AA for 5 min prolonged the microbiological shelf-life for 8 days compared to those of 0.5% AA and the controls. All treatments of AA increased the lag phase and GT of aerobic microorganisms.

This study was conducted to investigate the effects of pathogenic hngi(Fusarium culmorurn, Fusarium oxyspomm fsp. lycopersici, Fusaiurn solani, Rhizoctonia solani), pregermination time(0, 6, 9, 12, 18, 24, 48 hrs.) and temperature(12℃, 18C˚

This study was perfomed to investigate the differentiation of rabbit blastocysts microinjected with testosterone solution. A total of 140 mixed breed does was superovulated, synchronized and hand mated. The eggs were flushed from uterine horns between 65 and 89 hrs after mating. Testosterone was dissolved in 95% ethanol and diluted with PBS at the ratio of 1: 99. Final concentration of testosterone was adjusted to 1 pg /ml. 6~8 bias-tocysts were microinjected with 1~10 p Q of the diluted testosterone solution, and tranfer-red into the uterine horns of the synchronized recipients. When 140 donor does were treat-ed with a single does of 200 IU PMSG in combination with 100 IU RCG 48 hrs apart, 134 of them(97%) showed standing estrus. Ovarian responses of 117 does were examined following mating and the rate of ovulation was 11.23 i 1.20. Ova were recovered from donors between 65 and 89 hrs after mating. Recovery rates of ova were 37.5% and 42.2% of recovered ova were blastocysts. A total of 106 blastsocysts were microinjected with testosterone solution and transferred into the uterine horns of 15 synchronized recipient does. One of the recipients was pregnant and delivered 7 baby rabbits. The external genitalia of the young rabbits appered to be the same appearance as the buck entierly.

Acyl-CoA synthetase 4는 생쥐에 있어서 거의 모든 조직에서 발현하며 아라키돈산에 특이적인 효소이다. 아라키돈산은 세포막의 인지질로부터 cPLA2에 의하여 유리되고 cyclooxygenase-1, -2에 의하여 eicosanoid로 변환된다. 이렇게 생산된 prostaglandin과 같은 eicosanoid는 배란, 수정, 임신에 있어서 중요한 기능을 수행하고 있다. 그러나 세포막으로부터 유리된 아라키돈산은 acyl-CoA syntheta