PCR기법에 의한 소 수정란의 성감별

이해이,조성우,박기승,김용수,정구남,노수일,김용준 韓國受精卵移植學會 1999 한국동물생명공학회지 Vol.14 No.1

개 정자의 수정능력 검정을 위한 Hamster test의 이용 가능성

이해이,김용준,Lee Hae-Lee,Kim Yong-Jun 한국임상수의학회 1993 한국임상수의학회지 Vol.10 No.1

To investigate the availability of hamster test in assaying the fertilizing capacity of dog sperm and the effect of canine sperm motility on sperm binding and penetration, semen were collected from four dogs(three dogs had been proven to be fertile and one dog to be subfertile during the past two years) and then preserved in BWW(Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding) and ova penetrated by sperm(penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison between fertile dogs and a subfertile dog, the rate of sperm binding was higher in fertile dogs than the subfertile dog(p<0.01, p<0.05). The number of bound sperm per ovum was considerably higher in a fertile dog than the subfertile dog((p<0.01), and also difference of number of the bound sperm was obtained among the fertile dogs(p<0.01, p<0.05). The rate of penetration as well as the number of penetrated sperm per ovum was higher in the fertile dogs than the subfertile dog(p<0.01, p<0.05). In fertile dogs. the canine semen preserved at 4$^{\circ}C$ for 18 to 22 hours showed from 30 to 80% motility at Insemination, however, no difference in hamster test was obtained according to different degree of sperm motility. These results indicated that hamster test would be of avail in assaying the fertilizing capacity of dog sperm.

한우 수정란의 간이 동결을 위한 유리화 동결법에 관한 연구

이해이,김상훈,김용준 韓國受精卵移植學會 2014 한국동물생명공학회지 Vol.29 No.1

This study was carried out to study the survival rate of thawed Hanwoo embryos frozen by the slow-rate freezing or the cryotop vitrification method. Hanwoo cumulus-oocyte complexes were recovered from ovaries at a slaughter house, matured for 20~22 hours, fertilized with Hanwoo semen for 5~6 hours, and cultured for 7~9 days in 38.5℃, 5% CO2 incubator. For freezing, Day 7∼9 blastocysts were collected. Embryos for the slow-rate freezing were equilibrated in 1.8 M ethylene glycol (EG) with Dulbecco's phosphate-buffered saline (D-PBS). Programmable cell freezer was precooled down to -7℃, and the straw was seeded during 8 minutes-holding time, and was cooled to -35℃ at the cooling rate of 0.3℃/min, and then was plunged and stored in liquid nitrogen. Embryos for the cryotop vitrification were treated in TCM199 with 0.5 M sucrose, 16% EG, 16% dimethylsulfoxide (DMSO). Embryos were then loaded individually onto cryotop and plunged directly into liquid nitrogen. The survival rates of embryos frozen by these two freezing methods were evaluated at 12 to 24h post-thawing. The survival rates of frozen/thawed Hanwoo embryos by the cryotop vitrification method (56.86 ± 26.53%) were slightly higher than those by the slow-rate freezing method (55.07 ± 26.43%) with no significant difference. Using the cryotop vitrification and the slow-rate freezing of Hanwoo blastocysts on Day 7 following in-vitro fertilization (IVF) treatment, the survival rates of frozen/thawed Hanwoo embryos were 72.65 ± 18.3% and 79.06 ± 17.8%, respectively. The survival rates by the cryotop vitrification were higher than those by the slow-rate freezing on both Day 8 and 9 with significantly higher survival rate on Day 9 (p<0.05). Using the cryotop vitrification and the slow-rate freezing of Hanwoo embryos to compare between three different blastocyst stages, the survival rates of the blastocyst stage embryos were 66.22 ± 18.8% and 45.76 ± 12.8%, respectively with higher survival rate by the vitrification method (p<0.05). And the survival rate of expanded blastocysts was higher than those of early blastocysts and blastocysts in two freezing methods with significantly higher survival rate by the slow-rate freezing method (p<0.05).

칡소의 모색과 Melanocortin 1 Receptor(MC1R) mRNA: 3’-비번역 부위의 변이 및 발현

이해이,박재희,김종국 사단법인 한국동물생명공학회 2014 한국동물생명공학회지 Vol.29 No.3

The objective of this study was to determine the breed differences in the 3'-untranslated region (UTR) of MC1RmRNA, which may be used to distinguish Korean brindle cattle (Chikso) from other breeds. We investigated therelationship between the variation of 3'-UTR of the MC1R mRNA and coat color among different breeds and theKorean brindle cattle with different coat colors. MC1R mRNA expression levels were determined in accordance withthe coat color and hair colors of the tail. Total cellular RNA was extracted from the hair follicles of the tails inHanwoo, Korean brindle cattle, Holstein and Hanwoo × Holstein crossbred cattle. After cDNA synthesis, PCR wasperformed. Sequences of the 3'-UTR of MC1R mRNA were analyzed. The 3'-UTR of the MC1R mRNA from differentbreeds of cattle did not show any variations. There were no variations in the 3'-UTR of the MC1R mRNA in Koreanbrindle cattle with different coat colors. The levels of MC1R mRNA expression in hair follicles of the tail variedsubstantially among the Korean brindle cattle with different coat colors, except yellow coat color. Correlation betweenthe MC1R mRNA expression in the hair follicles of the tail and coat color may be present in the Korean brindlecattle, but not between the variations of 3'-UTR of MC1R mRNA and coat color. Further studies to determine theregulation of MC1R mRNA expression from the hair follicles of different coat colors will be beneficial in clarifyingthe role of MC1R in the coat colors of the Korean brindle cattle

후대의 육질등급이 우수한 한우 암소의 수정란 생산 및 이식에 관한 연구

이해이,박재희,김용수,김종국 한국동물생명공학회(구 한국동물번식학회) 2019 Journal of Animal Reproduction and Biotechnology Vol.34 No.1

These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), in-vitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

칡소의 모색 발현과 호르몬 변화와의 상관관계 분석 연구

이해이,박재희,김용수,김종국 사단법인 한국동물생명공학회 2015 한국동물생명공학회지 Vol.30 No.1

The aim of this study was to determine the relationship between the coat color appearance of Korean brindle cattleand the changes of relevant hormone levels that may affect the hair pigmentation during different stages of growthand maturation. In mature cattle, levels of both ACTH and DHEA in Korean brindle cattle with brown color weresignificantly higher than those with black color (p<0.05). Levels of α-MSH in Korean brindle cattle with whole brindle(≧50%) color were significantly higher than those with brown color (p<0.05). In calves of Korean brindle cattle at2 to 6 months, the concentration of estradiol was significantly higher in calves with whole brindle color than thosewith part brindle color (p<0.05), when the coat color was confirmed. After 6 month of coat color confirmation, levelsof testosterone and ACTH increased in calves with part brindle color and were significantly higher than those withwhole brindle color (p<0.05). In calves of Korean brindle cattle at 1 or 2 months, there were no significant differencesin hormone levels of estradiol, ACTH, DHEA and α-MSH between the calves with brindle color and brown color,except estradiol before brindle color appearance. Changes of relevant hormone levels at different stage of growth andmaturation may affect the pigmentation of coat during the development of cattle. In addition to the current studycorrelating the different coat colors with relevant hormone levels, investigation of the coat color associated genesexpressed in Korean brindle cattle may further clarify the mechanisms of coat color changes during their development

후대의 육질등급이 우수한 한우 암소의 수정란 생산 및 이식에 관한 연구

이해이,박재희,김용수,김종국 사단법인 한국동물생명공학회 2019 한국동물생명공학회지 Vol.34 No.1

These studies were conducted to evaluate developmental competence of follicular oocyte collected from the ovaries of Hanwoo cows with the high offspring meat quality (1++ and 1+ grade). Cumulus oocyte complexes from individual cows were matured, fertilized and cultured using protocols of in-vitro maturation (IVM), in-vitro fertilization (IVF) and in-vitro culture (IVC). The rates of blastocyst development from Hanwoo cows with the offspring meat quality grades of 1++ and 1+ were 18.6 and 21.2%, respectively. The rates of blastocyst development were 26.3, 20.7, 20.7, 17.2 and 31.2% from Hanwoo cows with the meat quality grades of 1++, 1+, 1, 2 and 3, respectively. Fiftyseven transferable embryos were recovered from 11 Hanwoo donor cows (5.2/head) with the high offspring meat quality grades of 1++ and 1+ in vivo, and the pregnancy rate after embryo transfer was 61.1%. In conclusion, these results suggest that in vitro embryo production from the ovaries of cows with the high meat quality grades using individual culture system can be used an efficient method for livestock improvement. In addition, for the successful industrialization of embryo transfer, conception rate should be improved.

소 정자의 운동성 향상에 따른 수정율 증진 연구

이해이,박재희,김용수,김종국 사단법인 한국동물생명공학회 2013 한국동물생명공학회지 Vol.28 No.1

This study examined the motility of either the unattached(upper) or attached(lower) Hanwoo sperm to bovine oviduct epithelial cell(BOEC) monolayers to determine whether there are any changes in their motility during co-culture. The cleavage and blastocyst development rate were compared among different preincubation methods in-vitro, after oocytes were fertilized in-vitro with Hanwoo sperm on BOEC monolayers. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 5 hours and 6 hours (p<0.05) of incubation, in sperm treatment medium without heparin and caffeine. The motility of frozen-thawed sperm in BOEC co-culture group was significantly higher than controls, especially at 3 hours (p<0.05) and 6 hours (p<0.01), in sperm treatment medium containing heparin and caffeine. The motility of the attached( lower) sperm was significantly higher than the unattached(upper) sperm during co-culture with BOEC at all times(p<0.01 or p<0.05), except for 6 hours. After Hanwoo oocytes were fertilized in-vitro with the sperm that had been co-cultured with BOEC in sperm treatment medium containing heparin and caffeine, we determined the cleavage and blastocyst development rate, according to the preincubation methods. Both the cleavage and blastocyst development rate from 2 hour preincubation group were the highest, but significant difference was not recognized. These results show that BOEC plays an important role on sperm hyperactivation related to capacitation regardless of heparin and caffeine in sperm treatment medium. However, oviduct epithelial cell had no significant effect on the development of embryos after in-vitro fertilization in the presence of added heparin and caffeine in sperm treatment medium.

특집-한글 번역 : 국제화 시대의 동양식 사유와 자각 -"네팔 삼부작"을 중심으로

해이수 단국대학교 한국문화기술연구소 2008 한국문화기술 Vol.5 No.-

Chak-Bup : The Buddhist Ceremonial Dance of Korea

Alan C. Heyman 한국무용연구회 1982 한국무용연구 Vol.1 No.-

윤달이 되면 절에서 행하여 지는 불교의식으로 큰 제를 볼 수 있다. 큰 제중에는 작법이 있는데 그중 “영산제 각 배제 수륙제” 가 있다. 큰 제에는 많은 제물이 바쳐져야 되기 때문에 많은 경비가 소요된다. 때문에 요즘은 윤달이 되어도 큰 제를 많이 볼 수가 없는데 주로 작법은 대처승 절에서만 행하여 지고 비구승은 하지 않는 것이 상례 이다. 비구승은 이를 무당굿에 해당되는 춤이라기 보기 때문에 행하여지지 않고 있다. 제의 작법중에는 4 가지의 층이 있는데 ①나비춤 ②바라춤 ③타주 ④법고 무가 있고 무용 반주에 는 2 가지의 범패와 조라치가 있다. 범패의 유래는 원래 인도의 노래이고 중국으로 전하여져서 다시 한국으로 전하여진 것으로 알려져 있다. 대부분 범패의 가사내용은 한문으로 되어있고 바라춤을 출 적에 반주하는 노래만 범어로 되어 있는데 바라춤을 출 적에 부르는 범패는 제목이 “천수경”으로 되어 있고 오늘날 바라춤의 제목이 “천수 바라”로 전해 오고 있다. “조라치”는 악기의 반주다. 조라치의 반주때 연주되는 악기는 호적,나팔, 소라, 북, 징, 제금이다. “조라치” 를 연주하는 악사들의 복장은 노란색이고 옛날 국군악으로 연주되었다. 불교제는 시작할때에 절 마당에 스님과 악사 등 여러 사람들이 돌아가면서 시주하는 식으로 이어 나가는데 부정을 없애기 위하여 하는 방법이다. 이때의 연주되는 가락은 "대취타”가 연주되는데 절마당을 빙빙 돌면서 한두사람의 스님이 가운데 에서 나비춤을춘다. 이때 악사들이 부르는 가락은 “염불”이고, 스님들이 부르는 범패 제목이 “도량계 "가 있는 데 작법중 “나비춤"을 제일 중심으로 추는 춤이다. 나비춤 : 의상은 머리에 높은 고깔을 쓰고 소매가 넓고 가슴엔 빨간색의 가사가 있다. 어깨와 등허리엔 5 가지 색깔의 리본이 매어져 있는데 5 방향을 표시하는 것이다. 나비춤의 사위중 제일 으뜸되는 것은 시작할때에 왼발의 발꿈치를 오른발 발꿈치에 살짝 올려 놓고 무릎은 약간 구부리고 두발을 움직이면서 제자리에서 빙빙 돌면서 제자리에 되돌아 오면 다시 왼발의 발꿈치를 오른발의 발꿈치에 원형 대로 올려 놓는다. 한국의 고전춤 중 작법과 불교의식의 춤만이 앞에서 말한 춤사위가 있다. 나비춤의 사위는 여러 가지가 있는데 앞에서 말한 춤 사위는 제일 중심되는 춤 사위이고 둘째는 양손을 벌리고 살짝 나비의 모습을 연상하면서 양손을 오르락 내리락 살짝 살짝 움직인다. 타주 : 의상은 나비춤 의상과 똑 같고 두손으로 막대기를 잡고 2 사람의 스님이 양쪽으로 서서 팔정도를 가운데 놓고 돌아가면서 치는데 이 춤사위는 영산제 의 식당작법 중에 행하여진다. 법고 : 영산제의 식당작법에 연주하는 큰 북춤이다. 큰 북 앞에 스님 한사람이 북을치고 춤을 추는데 조라치는 시작할때 는 천천히 하고 계속될 수록 빨라진다. 한국 전통 무용중에 법고는 제일 어려운 춤이다. 법고무를 식당작법에 연주하는 이유는 큰절에서의 점심 저녁을 알리는 종소리로 사용하기 때문이다. 바라무 : 2 개의 큰 바라를 양손에 들고 머리위에 손을 올려 머리위에서 손을 앞뒤로 혼들면서 바라를 돌리면서 몸전체를 돌린다. 바라는 무게가 많기때문에 특히 천수바라는 팔에 힘이 많이 가는 춤이다. 영산제 중에서 제일 재미나고 구경거리가 많은 것은 식당작법이다. 많은 경비가 소요되기 때문에 요즘 구경하기가 힘들다. 식당작법을 한번 구경하면 영원히 머리속에서 떠나지 않는다.