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돼지수정란의 Compaction 양상에 따른 착상전 배발달 양상
구덕본,민성훈,박흠대 韓國受精卵移植學會 2010 한국동물생명공학회지 Vol.25 No.3
Embryonic compaction is essential for normal preimplantation development in mammals. The present study was to investigate the effects of compaction patterns on developmental competence of pig embryos. The proportion of blastocyst formation derived from compacted morula was higher than those of compacting and pre-compacting morula (P<0.01). Nuclei numbers of inner cell mass (ICM), trophectoderm (TE), and total of blastocysts derived from compacted group were also superior to those of compacting and pre-compacting groups (P<0.05). Then, compaction patterns, developmental ability and structural integrity were compared between mono- and poly-spermic embryos. The rate of compacted morula in mono-spermic embryos was higher than that of poly-spermic embryos (P<0.05). Especially, the rate of blastocyst formation derived from compacted embryos in mono-spermic embryo group was higher than that of poly-spermic embryo group (P<0.05), although no difference was detected between the two groups in the structural integrity. Finally, we confirmed that beta-catenin was differentially expressed according to compaction patterns in morula and blastocyst stage embryos. In conclusion, our results suggest that the compaction patterns during preimplantation development play a direct role in developmetal competence and quality of pig embryos.
김민지 ․ 구덕본 대구대학교 산업기술연구소 2018 産業技術硏究 Vol.29 No.2
Production of transgenic animals are being used in various fields such as commercial application, human diseases model and drug discovery. One of these animals production method of transgenic animal is direct transfer by pronuclear microinjection. However, the production efficiency of transgenic animals using the pronuclear microinjection is remarkably low. In addition, appropriate conditions such as improving the embryo developmental competence and high quality of blastocysts for the production of transgenic embryos are being required, but studies for these process have not been reported. Therefore, this study investigated the embryonic developmental competence and quality of blastocysts after pronuclear microinjection by using cytochalasin B (CB) or/and melatonin. Cytochalasin B (CB) is important to play role in microinjection by making smoother of the zygote cytoplasm, as well as melatonin is known to improve the embryonic developmental competence and embryos quality. First, we performed the microinjection using injection medium with 10 µg/ml CB and cultured the microinjected zygotes in 0.1 µM melatonin treated within culture medium. Next, we confirmed the DNA damage of microinjected blastocysts using immunofluorescence staining with RAD51 (DNA repairs detection protein) and H2AX139ph (DNA damage detection protein). CB treated group significantly increased blastocyst development rate compared with CB non-treated group (P < 0.05; 25.0 ± 7.2% vs. 17.0 ± 1.3%). After microinjection under the CB treatment, blastocyst development rate and formation of expanded blastocysts were higher in melatonin treated group than those of melatonin non-treated group (29.7 ± 8.3% vs. 24.4 ± 7.7%). In addition, melatonin treated group increased the numbers of RAD51 positive cells compared with non-treated group (P < 0.05). Taken together, these results suggest that CB and melatonin treatment have positive effects for improving the production efficiency of transgenic embryos.