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체외수정 및 미세조작에 의한 가축배의 생산과 효율적 이용에 관한 연구 6. 소 핵이식배의 체외배양 , 동결보존 및 성판정에 관한 연구
정영채,김창근,윤종택,나광빈,오성종,이종완,김흥률,김광식,박선애,유영아 ( Y . C . Chung,C . K . Kim,J . T . Yoon,G . B . Luo,S . J . Oh,J . W . Lee,H . R . Kim,K . S . Kim,S . A . Park,Y . A . You ) 한국축산학회 1997 한국축산학회지 Vol.39 No.3
This study was carried out to establish an effective method for the improvement of subsequent development after nuclear transfer(NT) in bovine embryo, for the extension of utility through freezing and sexing of NT embryos. The rates of cleaved and developed embryos upto morula-blastocyst stages after the NT in in vitro produced embryos were 60.6 and 3.7%, respectively. Whereas, in in vivo embryos, slightly higher developments were obtained(70.8 and 5.6%), respectively. No difference was found in embryonic development upto morulablastocyst stages among the different levels of EGF added to the serum containing medium. The rates of cleaved and developed embryos upto morula-blastocyst were higher when electrostimulation was performed either before fusion or after addition at 21h IVM. The survival rate of frozen NT embryos was higher in late morula than that of earlier embryos. PCR-based sexing of NT embryo demonstrated that higher proportion of embryos was male(1.9:1), instead of normal ratio(1:1).
한국재래산양에서 계절이 정소기능, 정자의 내동성 및 수정능력에 미치는 영향 I. 정액성상과 정자의 내동성 및 난자침입능력의 계절적 변화
김창근,정영채,김광식,윤종택,이장희,정영호,최선호,김흥률,김수,권처진,Kim, C.K.,Chung, Y.C.,Kim, K.S.,Yoon, J.T.,Lee, J.H.,Chung, Y.H.,Choi, S.H.,Kim, H.Y.,Kim, S.,Kwon, C.J. 한국동물번식학회 1994 Reproductive & developmental biology Vol.17 No.4
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.
한국재래산양에서의 계절이 정소기능, 정자의 내동성 및 수정능력에 미치는 영향 II. 혈청내 Testosterone수준의 계절 및 하루중 변화
김창근,정영채,김광식,김수,권처진,한기영,윤종택,정영호,이장희,Kim, C.K.,Chung, Y.C.,Kim, K.S.,Kim, S.,Kwon, C.J.,Han, K.Y.,Yoon, J.T.,Chung, Y.H.,Lee, J.H. 한국동물번식학회 1994 Reproductive & developmental biology Vol.17 No.4
Seasonal and diurnal variation in serum testosterone was measured by radioimmunossary in 3 mature Korean native male goats. Blood was collected at 3-h intervals for 24h in each season. Mean levels of 4 seasons ranged from 0.82 to 2.06ng/ml and no difference was among seasons, although level in fall and winter was slightly higher. Diurnal level tended to increase at daytime in all season and when diurnal variation in level was compared to daily mean of each season, level of daytime greatly(P<0.05) increased in summer. However, variation between light and dark period in other seasons did not show significant difference. Daytime increase of testosterone was found 3 to 4 after dawn and level increased 1 to 2h ahead in summer and fall than that in others. These results in serum testosterone suggest that there was no great seasonal variation in endocrine function of goat testis.
미경산돈에서 교배전 자궁내 정액항원 주입에 의한 산자수 증가에 관한 연구
한기영(K . Y . Han),김창근(C . K . Kim),윤종택(J . T . Yoon),김형태(H . T . Kim),정영채(Y . C . Chung),서경덕(K . D . Seo),연정웅(J . W . Youn),이용우(Y . W . Lee) 한국축산학회 1988 한국축산학회지 Vol.30 No.9
This experiment was conducted to evaluate effect of a single intrauterine infusion of semen or seminal plasma at estrus before mating on subsequent litter traits of crossbred gilts. In addition, presence and level of early pregnancy factor activity (rosette inhibition titre, RITZ in serum of treated gifts and in vitro immunosuppressive activity of these antigens from boars used were tested immunologically to support the results in this experiment. The results obtained were as follows: 1. Semen-infused gifts significantly had a larger litter size (11.4) at birth than controls (9.6), but no significant differences were detected between others. 2. There was a tendency toward increased total pigs born alive, litter weight at birth and number of pigs suckling in antigen-infused gifts, but the differences between treated-gifts and controls were not statistically significant. 3. EPF activity (RIT) before and after mating in previously infused-gifts was 10.7±2.8 in the serum of nonpregnant gifts and 19.3±3.8 in the serum of pregnant gifts. RIT $gt; 14 could be considered evidence of presence of EPF in pregnant gifts. 4. Seminal plasma infusion before mating induced on obvious rise of RIT. RIT at 24 h and 48 h after mating was much higher (18-23 and 21-22) :in semen-and seminal plasma-infused gifts than in controls. RIT at 24 h after mating was found to be positively correlated (r=0.84) to litter size born in 6 gifts. 5. Dialyzed seminal plasma from 4 boars was significantly immunosuppressive at a concentration of 500 ㎍/㎖ than controls and degree of immunosuppression varied among individuals when the concentrations was under 500 ㎍/㎖. Undialyzed semen samples from 3 boars resulted in loss of immunosuppressive activity. These results indicate that infusion of gifts with semen at estrus before mating can be improved reproductive efficiency because of more litter size at birth and sukling pigs than controls and that these antigens had immunosuppressive potential of antilymphocyte serum and immunosuppressive activity to porcine lymphocytes that generate an immune response.
김창근(C . K . Kim),정영채(Y . C . Chung),윤종택(J . T . Yoon),최선하(S . H . Choi),류범용(B . Y . Ryu),정광조(K . J . Chung),김흥율(H . R . Kim),송해범(H . B . Song) 한국축산학회 1990 한국축산학회지 Vol.32 No.1
Effects of adding 10 to 20% fetal calf serum(FCS) and estrous cow serum(ECS), and hormones(GTH or GTH plus E₂) to maturation medium on in vitro maturation(IVM) and in vitro fertilization(IVF) of bovine follicular oocytes and effects of sperm treatment and bull on IVF and embryo development were studied. Finally, developmental potential these in-vitro fertilized embryos at 2-to 8-cell stage(44h postinsemination) was investigated under in vivo(rabbit oviduct) and in vitro(Ham`s Fl0 plus 10% FCS with or without bovine oviduct epithelial cells, BOEC) 4 day-culture system. 1. Addition of 10 to l5% FCS to mKRB and DM(-BSA) and of 20% FCS to Ham`s F10 increased(65%) IVM, respectively. IVM of these media supplemented with hormones was higher(70∼72%) than that of media without hormone. Addition of ECS showed higher IVM at 10% level for mKRB and 10 to 15% for TCM199 and the highest IVM(88%) was obtained in TCM199 containing 10% ECS and LH. 2. Higher IVF rate (based on formation of both pronuclei) in Exp I was obtained from oocytes matured in mKRB plus 10% FCS and HIS-sperm treatment. In Exp 2. IVFrate (based on cleavage. 2-3 cells) of oocytess matured in TCM199 was higher than in other media and IVF using fresh semen was significantly higher but was similar between sperm treatments. In Exr 3, IVM in TCM199 containing 10% ECS and LH and IVF by C+H-sperm treatment significantly increased early cleavage(2-to 8-cell). 3. In vitro-acrosome reaction(AR) among individual bulls in Exp 1 ranged from 16 to 45% and bull group having lower AR showed low IVF rate than group having higher AR. In Exp 3, cleavage rate by sperm treatment with both chemicals(C+H) was higher than that by H alone. 4. In IVF with in vivo-capaciated sperm, development from 2-to 3-cell to above 8-cell was higher under in vivo culture(54.5%) than under in vitro culture(31.3%) and higher in oocytes matured in TCM199 plus to ECS. In IVF with in vitro-capacitated sperm, development into above 8-cell stage was different between initial stage(2-to 3-and 4-to 8-cell) of embryos(26.7% : 41.7%) and significantly lower under in vitro culture from 2-to 3 cell stages(l0% : 29.4%). Under in vitro co-culture with BOEC, embryo development from 4-to 8-cell was significantly improved(53.3%) and 3(20%) of 15 cultured embryos developed into morula. The present results indicate that addition of ECS and hormones can improve IVM, IVF and development of bovine follicular oocytes, sperm factor may be important for IVF and further development, and co-culture with BOEC is an effective means for development into above 16-cell stage.