간행물
한국동물생명공학회지 (구 한국수정란이식학회지) KCI 등재 Journal of Animal Reproduciton and Biotechnology
- 발행기관 한국동물생명공학회(구 한국수정란이식학회)
- 자료유형 학술지
- 간기 계간
- ISSN 2671-4639 (Print)2671-4663 (Online)
- 수록기간 1983 ~ 2024
- 주제분류 자연과학 > 생물공학 자연과학 분류의 다른 간행물
- 십진분류KDC 527DDC 636
권호리스트/논문검색
Vol. 9 No. 2 (1994년 8월) 8건
1.
1994.08
구독 인증기관 무료, 개인회원 유료
The suitable electric stimulation is essential for activation and fusion of oocytes before or after nuclear transplantation The present study was undertaken to determine the optirnal condition for the parthenogenetic activation of in vitro rnatured(IVM) bovine oocytes by electric stimulation. Different direct current(DC) electric voltage of 1.0, 1.5 and 2.0 kV/cm and pulse duration of 30, 60 and 120 sec were applied to the JVM nocytes in 0.3 M mannitol solution containing each 100 M CaCl and MgCl. IVM occytes at 24, 28 and 32 hours Post-maturation(hpm) were also electrically stimulated at 1.5 kV /cm, for 60 sec. The stimulated nocytes were then co-cultured in TCM-199 solution containing 10% fetal calf serum with bovine oviductal epithelial cells for 7~9 days in a 5% incubator at 39 ~ Their activation and in vitro development to morula and blastocyst were assessed under an inverted microscope. The higher activation rates 62.8 and 63.4% and in vitro de- velopment rates to morula and blastocyst 5.1 and 10.9% were shown in the oocytes stimulated at the voltage of 1.0 and 1.5 kV/cm than 2.0 kV/cm, respectively. No signifi- cantly(P<0.05) different activation rate was shown in JVM oocytes stimulated for 30, 60 and 120 sec, but developmental rates to morula and blastocyst was significantly(P<0.05) higher in the oocytes stimulated for 30 sec(6~3%) and 60 sec(10~0%) than 120 sec(0~ 0%). The aged oocytes at 28 and 30 hpm showed significantly(P<0.05) higher activation rates(72~7 and 79.7%) than the oocytes at 24 hpm(50~9%)~ Also, their developmental rates to morula and blastocyst were significantly(P<0.05) higher in the nocytes at 28(14.3%) and 32 hpm(15.9%) than 24 hpm(3.6%). From these results, it can be suggested that the optimal electric stimulation for IVM bovine occytes is a DC voltage between 1.0 and 1.5 kV/cm, pulse duration of 30 or 60 sec, and the optimal age of IVM oocytes for electric activation is at 32 hpm.
4,000원
2.
1994.08
구독 인증기관 무료, 개인회원 유료
This study evaluated the influence of cell stage of donor nucleus on nuclear injection, electrofusion and in vitro development in the rabbit to improve the efficiency of nuclear transplantation in the rabbit. The embryos of 8-, 16- and 32-cell stage were collected from the mated does by flushing viducts with Dulbecco's phosphate buffered saline(D-PBS) containing 10% fetal calf serum(FGS) at 44, 54 and 60 hours after hCG injection. The blastorneres separated from these embryos were used as donor nucleus. The ovulated oocytes collected at 14 hours after hCG injection were used as recipient cytoplasm following removing the nucleus and the first polar body. The separated blastomeres were injected into the enucleated oocytes by micromanipulation and were electrofused in 0.28 M mannitol solution at 1.5 kV /cm, 60 sec for three times. The fused oocytes were cocultured with a monolayer of rabbit oviductal epithelial cells in M-199 solution containing 10% FGS for 72~120 hours at 39 in a 5% incubator. The cultured nuclear transplant embryos were stained with Hoechst 33342 solution and the number of cells were counted by fluorescence microscopy. The successful injection rate of 8-, 16- and 32-cell-stageblastomeres into enucleated oocytes was 86.7, 91.0 and 93.9%, respectively. The electrofusion rate of 8-, 16- and 32-cell-stage blastomeres with enucleated oocytes was 93.3,89.3 and 79.0%, respectively. Development of blastomeres to blastocyst was similar with 8-,16- and 32-cell-stage donor nuclei(26.2, 25.8 and 26.6%, respectively, P<0.05). The mean number of cell cycle per day during in vitro culture in nuclear transplant embryos which received 8-, 16- and 32-cell- stage nuclei was 1.87, 1.81 and 1.43, respectively.
4,000원
3.
1994.08
구독 인증기관 무료, 개인회원 유료
This experiment was carried out to produce cloned aniraals by nuclear transplantation in rabbits. The ovulated oocytes were collected from the oviducts between 14 and 15 hours after hGG injection. The denuded oocytes were used as nuclear recipient cytoplasm following enucleation by micromanipulation. The blastomeres separated from the 8-cell embryos were used as nuclear donor. The nucleated oocytes receiving a blastomere in the perivitelline space were electrically fused in the 0.28 M mannitol solution at 1.5 kV /cm, 60sec for three times. The nuclear transplant embryos which were used and developed to 2- to 4-cell stage in vitro were transferred into the oviducts of synchronized recipient does. A total of 64 nuclear transplant embryos were transferred to 7 recipient does and produced three offspring(4.7%) from a foster mother 31 days after embryo transfer.
4,000원
4.
1994.08
구독 인증기관 무료, 개인회원 유료
This experiment was arried out to investigate the development of ea4y rabbit embryos in vivo. Twenty-six New Zealand White does were superovulated by treatment with PMSG(Intervet Co; I. M single injection, 150. U./rabbit) followed 3 day later by simultaneous I.V injection of 100 I.U HCG (Intervet Co, )and natural service with fertile male. All of does was killed at the specific times (24, 27, 30, 36, 42, 50 and 93 h post-hCG) to find out the early embryonic development in vivo respectively. Embryos at the specified stages of development were obtained at the following times after injection of hCG; one-ceH at 24 h, two-cell at 24~27h, four-cell at 27~36 h, morulae at 50 h and early blasto-cyst at 93 h and expanded or hatching blastocyst at 144 h. Number of embryos recovered per rabbit superovulated was 26.1 and average of recovery rate was 83.7%. The results suggest that superovulation was efficient for the increase of embryo number in rabbits, and as shown in results, asynchronous cleavage was prevalent among the recovered embryos.
4,000원
5.
1994.08
구독 인증기관 무료, 개인회원 유료
The present experirnents on cryopreservation were carried out to investigate effect of solution toxicity, equilibration time and cell stages on the post-thaw survival of mouse morulae and blastocyst embryos cryopreserved by vitrification in EFS solution. The mouse embryos were exposed to the EFS solution in one step at room temperature, kept in the EFS solution during different period for toxicity test, vitrified in liquid nitrogen and thawed rapidly. After the mouse morulae embryos were exposed to EFS solution for 2 and 5 ruin. at room temperature and then they were washed in 0.5 M sucrose solution and basal mediurn(D-PBS + 10% FCS), they were cultured to examined cryoprotectant toxicity induced injury during exposure, most of embryos developed to expanded blastocysts(100 and 90.0%). However, when the exposure time was extended to 10 and 20 min, these development rates dropped dramatically in 10 ruin. (75.0%) and 20 ruin. (4.5%), respectively. When the compacted morulae were vitrified in EFS solution after equilibration for 2 and 5 min, the embryos have developed to normal blastocyst following thawing, washing and culture processes was 89.3 and 89.6%. However, when the exposure time was expanded to 10 ruin, this survival rate dropped to 68.8%. When the blastocyst were vitrified in EFS solution after equilibration for 2, 5 and 10 minutes, the survival rate of embryos which developed to normal blastocyst following thawing and culture processing were 58.5, 46.7 and 22.4%, respectively. The optimal time of equilibration of mouse morula and blastocysts in EFS solution seemed o be 2 and 5 ruin.
4,000원
6.
1994.08
구독 인증기관 무료, 개인회원 유료
This research was conducted to obtain the basic information on the cell block phenomenon occuring during early development in vitro of mouse embryos. Early embryos were recovered at 3h post-hGG injection(hph). Various chemicals (EDTA, EGTA, DTPA, MA and PRA) were tested to examine the effects of them on the overcoming the 2-cell block phenomenon. One hundreds M of the chelating agents were added to the M16 medium containing embryos. The treated embryos were worked and transferd to fresh M16 medium after 1, 3, 6 and 12h of treatment. Development was examined at 58 and l2Oph injection, respectively. 44.7~68.9% of the treated embryos developed to 4-cell stages at 58hph. Only 17.6~60.3% of the embyos developed upto blastocyst at l20hph. Whereas control embryos showed slightly lower development in M16 medium alone (38.9~42.4%, 4-cell and 3.8~65.5%, blastocyst). Three mitogenic agents were tested. 51.6~63.8% and 43.4~48.1% of embryos developed up to 2-cell and blastocyst stage, respectively when treated in 5 g PHA-M Imi for 5 min, 1, 3 and 6h subsequently cultured in fresh M16 medium. Control embryos only showed 38.8% for 4cell and 5.9% fo blastocyst at 58 and l2Ohph, respectively. 100M PMA was also beneficial for the 2-cell block. Showing better development them that of control (42.4 vs 57.9~59.4% 4cell and 5.9 vs 25.0~55.6% blastocyst, respectively. However 1M butyric acid was toxic to early embryos, thus arresting further development. These result indicate that either chelating or mitogenic agents could be used to overcome the "in vitro 2-cell block" occuring during early development in vitro of ICR embryosCR embryos
4,000원
7.
1994.08
구독 인증기관 무료, 개인회원 유료
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.
4,000원
8.
1994.08
구독 인증기관 무료, 개인회원 유료
This experiment was carried out to clarify the pedigree identification from blood typing of 301 Hoisteins in National Animal Breeding Institute(N.A.B.I.). Twenty kinds of standard reagent standardized by Insternational Society for Animal Blood Group Research provied from KNC improvement center, N, L, C, F. were used as the reference reagents in this study. The highest frequency of antigenic facfors was obtained from Xin blood typing of 301 Holsteins. The frequency of X was 0.714.In A blood system, four kinds of phenogroups were observed. The gene frequencies of Al and Z' phenogroups were equally 0.027.This frequency was greatly lower than those of breeds of Southern European and Zebu cattle. In B blood Systern, nineteen kinds of blood type were appeared. The appearance frequency of Gx blood type was 0.259, whish was higher than the others. In C blood system, thirty kinds of blood type were observed. The appearance frequency of X blood type was the highest(0.189). In F blood system, three kinds of alleles were detected. The gene frequency of F allele was higher than that of V(0.105). However, the frequency of F allele(0.327) was greatly lower than that of "- /- " allele. In S blood system, twelve kinds of blood type were appeared and showed sirnilar appearance frequencies except " - / - " allele. From the results of the pedigree identification from 8 sires and 28 progenies of them, the accuracy of pedigree identification was 92.9%.ification was 92.9%.
4,000원
