난자의 성숙과정과 노화에 관한 이해는 인공수정과 체외수정 최적기를 판단하기 위하여 가장 중요한 연구내용으로 알려져 있다. 이러한 기작은 번식 호르몬들에 의하여 조절되는 것으로 알려져 있으나 난자 세포질 변화에 관한 내용은 잘 알려져 있지 않다. 본 연구에서는 산화질소물(nitric oxide, NO)이 난자 성숙과정에서 증가하는 것을 밝혔으며 난자의 미성숙단계(germinal vesicle stage, GV)와 난자핵막붕괴단계(germinal vesicle breakdown, GVBD) 및 성숙완료단계(metaphase II, MII)단계에서 생산되는 NO의 양을 비교하였다. 또한, 난자를 체외에서 배양할 때, MII단계로 성숙되지 않는 성장 단계의 난자에서는 NO의 증가 현상을 관찰할 수 없었고, 세포질이 불균일한 노화된 난자에서는 NO가 증가된 상태로 유지되는 특성이 있음을 밝혔다. 이러한 결과는 NO의 작용이 난자의 성숙과정과 난자 노화과정에서 중요한 기능을 담당하고 있음을 보여주고 있다.

엘크 수사슴의 체중 성장곡선 및 일당증체량을 추정하기 위하여 국립축산과학원 가축유전자원센터에서 사육중인 엘크 수사슴 183두의 체중-일령 자료 1,509개에 대해 비선형 회귀식인 Gompertz와 Logistic 모형을 적용하였다. 추정된 엘크 수사슴 성장곡선 모수인 성숙체중(A), 성장비율(b)과 성숙률(ｋ)은 Gompertz 모형의 경우 각각 335.5±1.415kg, 2.18±0.045와 0.0025±0.00005였고, Logistic 모형에서는 330.5±1.337kg, 5.46±0.209와 0.0355±0.00009였다. 적합도 검정을 위한 결정계수(R2)는 두 모형에서 큰 차이가 없어 두 모형 다 적합한 것으로 나타났다. 다만, 실제 측정값과 추정값에 대한 차이인 평균제곱근 오차 값이 엘크 암사슴에 비해 크게 나타났으며 따라서 엘크 수사슴의 낙각 및 번식기 체중 증감을 고려한 성장곡선 모형에 대한 추가적인 연구가 필요할 것으로 판단된다. 엘크 수사슴에서 Gompertz 모형으로 추정된 함수식은 f(t)=335.5Xe^-2.18e^-0.0025t였고, Logistic 모형의 경우 F(t)=330.5X(1+5.465e^-0.00355t)^-1였다. 각 모형에서 제1차 도함수로 유래된 일당증체량 곡선은 Gompertz 모형의 경우 F'(t)=0.0025Xf(t)X1n(335.5/f(t)), Logistic 모형의 경우f'(t)=0.0036Xf(t)X[1-f(t)/330.5]였다. 이 함수식을 통해 도출된 엘크 수사슴에서 최대 증체량을 나타내는 시기인 315~478.3일령, 123.4~165.3㎏ 도달 전후에는 보다 세심한 사양관리가 필요할 것으로 보인다. 본 연구에서 도출된 엘크 수사슴의 성장곡선 모수 추정값, 변곡점 및 변곡점 도달일령 등은 사슴산업 종사자들이 사양관리 및 육종계획을 수립하는 데에 도움이 될 수 있을 것으로 사료된다.

Ovarian folliculogenesis and the production of fertilizable oocytes depend on gap junctional intercellular communication within both the developing and the mature follicle. Gap junctions connect oocytes with granulosa cells and granulosa cells with each other. Various nutritional bio-molecules are known to be transferred to the growing oocyte from the granulosa cells via gap junction. Signals that regulate meiotic maturation of fully-grown oocytes pass through the oocyte-granulosa cell gap junctions. Gap junctions also play a critical role in regulating uterine blood flow, contributing to the maternal recognition and also implantation during pregnancy. Due to the challenge of various stressors the in vitro embryo developmental potentials are still suboptimal compared to in vivo. To identify the molecular mechanism of these stressors and to improve the existing embryo developmental potentials, the singlet oxygens quencher lycopene was added to the culture media to counterbalance the oxidative damage caused by ROS. In this study, we have patterned connexin like Cx43, Cx37, Cx32 and Cx26 at protein and transcription level during follicular growth, atresia and blastocyst stage by using immunohistochemistry, conventional PCR and RT-qPCR. Lycopene (0.2 μM) significantly (P < 0.05) increased the gap junctional communication protein (connexin) expression of Cx43, Cx37, Cx32, Cx26 as compared to the control group at both transcription and translation level during follicular growth, atresia and blastocyst stage. Lycopene potentiates ovarian folliculogenesis, provides the production of fertilizable oocytes and improved embryo developmental capabilities by increasing gap junctional intercellular communication.

Transglutaminase (TGM2) belongs to a family of cross-linking enzymes responsible for catalyzing Ca2+-dependent acyl-transfer reactions between the substrate proteins. TGM2 is a cytosolic protein that has also been observed in the nucleus and can be expressed to the cell surface or extracellular matrix. Despite ubiquitous expression, its functions are poorly understood and still need to be elucidated. Moreover, there is no clear data regarding the role of transglutaminase in mammalian oocytes. So, in this study, we have patterned the transglutaminase 2 (TGM2) and anti-N epsilon gamma glutamyl lysine (AB424) activity in heat stressed mouse oocytes. We have collected mouse oocytes from the (6–9 weeks old) mouse and in vitro matured for 20 h. Immunocytochemistry was performed to checked the transglutaminase 2 (TGM2) and anti-N epsilon gamma glutamyl lysine (AB424) activity after 6 h of heat stress (HS) at 39.1 ℃. Both TGM2 and AB424 expression were significantly (P < 0.05) higher compared to control when oocytes were subjected to HS at 6 h of IVM at 39.1 ℃. Our hypothesis is that TGM2 and AB424 activity may be correlated with the cellular regression and also involvement in apoptosis. We hope that, our study will help to elucidate the normal function of mouse oocyte and also identification of the principal proteins as well as the pathogenic mechanism of altered physiology. These results suggest that the nuclear accumulation of the transglutaminase may play an important role in nuclear remodeling during folliculogenesis and early embryonic development

The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58 ± 8.31 and 13.25 ± 7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75 ± 1.98 vs. 8.23 ± 6.07, P<0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P<0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p<0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

The objective of this study was to investigate the result of in vivo embryo collection and pregnancy rate after embryo transfer using sex-sorted sperm of Korean brindle cattle. Donor Korean brindle cattle superovulation treated by decreasing dose of FSH injection. Embryos were recovered on 7 days after the third artificial insemination. Control group semen straw used artificial insemination contained 20 million sperm. Sex-sorted semen straws contained 4 million sperm or 10 million sperm. As for the result of the recovery of the in vivo embryos derived from sex-sorted sperm, the number of transferable embryos was significantly highly recovered to be 6.20±2.28/donor from the control group and was significantly lowly recovered to be 1.57±1.72/donor from the group treated at a sperm concentration of 10×106 (p<0.05). The number of unfertilized embryo was 0.8±1.30/donor in control group which was significantly lower than the group treated at a sperm concentration of 4×106 (p<0.05). However, there was no significant difference in the number of undeveloped ova between control and treatment groups. Pregnancy rate after embryo transfer was shown to be 35.00% in control group and 12.50% in treatment group. The karyotype analysis of the calf derived from sex-sorted sperm resulted in a similar chromosomal distribution pattern (2n=60, XX) compared to those of common Korean native cattle.

본 시험은 우리나라 전통한우인 칡소, 흑우, 백우의 생시, 이 유시, 12개월령, 18개월령, 24개월령의 체중과, 체형형질을 비 교분석하기 위하여 수행하였다. 공시축은 농촌진흥청 국립축 산과학원 가축유전자원센터에서 2011~2014 년도 까지 사육된 칡소 10두, 흑우 15두, 백우 7두를 이용하였다. 개월령별 체중 변화를 살펴보면, 생시 ~ 12개월령 체중은 흑 우가 가장 높았고 칡소는 가장 낮은 경향을 보였고, 18개월 령 ~ 24개월령 에서는 백우가 가장 높게 조사되었고 칡소가 가 장 낮게 조사되었다. 전통한우 24개월령 체형형질 측정치가 암소는 같은 개월령 의 황우와 비슷하게 추정되었지만, 수소는 26개월령 황우 거 세우와 비교하여 큰 차이로 낮게 추정되었다. 이러한 결과는 황우 수소의 개량효과를 시사하는 결과라고 사료된다. 암소의 번식관련 형질인 고장, 좌골폭과의 상관관계를 살펴 보면, 칡소는 체중, 체장, 요각폭과 높은 상관관계를 보였고, 흑우는 체고, 십자부고, 요각폭과 높은 상관관계를 보였으며, 백우는 모든 형질에서 높은 상관관계를 보였다. 수소의 강건 성관련 형질인 흉위, 흉폭과의 상관관계를 살펴보면, 칡소는 체장, 고장, 요각폭, 곤폭과 높은 상관관계를 보였고, 흑우는 체중, 십자부고, 고장, 요각폭과 높은 상관관계를 보였으며, 백 우는 좌골폭을 제외한 모든 체형형질에서 높은 상관관계를 보 였다. 이러한 상관관계를 고려하여, 체중 및 체형형질의 중요 한 간접선발 지표로 암소는 요각폭, 수소는 고장을 활용할 수 있을 것으로 사료된다. 종합적으로 칡소, 흑우, 백우에 대해 우리 고유유전자원으로 서의 보존의 한계를 극복하고 향후 이들의 산업적 이용을 고 려할 때 무엇보다도 개체 수 확보가 가장 시급한 과제이다. 이를 위해서 학계, 연구계 및 산업계가 연계된 체계적이고 다 양한 연구와 이들에 대한 구체적인 개량방향 및 목표설정 등 이 먼저 이루어 져야 할 것으로 사료된다.

The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was 20×106/straw. Sexed X frozen semen with 2×106 cells or 4×106 cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was 24.9 ± 7.31% and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between 2×106 cells or 4×106 cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with 2×106 cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

림프구는 면역기능의 중심적인 역할을 담당하는 세포이고, 운동에 의해 변화하고 노화에 의해 기능이 저하하는 것으로 잘 알려져 있지만, 고령자를 대상으로 운동에 의한 림프구의 변동에 대한 연구는 지금까지 미흡한 실정이다. 그래서 본 연구에서는 운동훈련이 고령흰쥐의 림프구에 미치는 영향에 대해 조사하였다. 8주간의 운동처리가 비장세포 중 그리고 장간막 림프절의 림프구에 미치는 영향을 분석했다. 비장세포중의 림프구에 대해서는 헬프-T 림프구, 세포상해성 T 림프구, 그리고 보조자극 신호 수용체 발현 T 림프구의 비율이 운동처리구에 있어 유의적으로 높은 수치를 나타냈다. 한편, 비장세포중의 B 림프구 및 장간막 림프절중의 림프구에 대해서는 두 처리구 간에는 유의적인 차이는 보이지 않았다. 이상의 결과들로부터, 운동처리구가 흰쥐 비장세포중의 T 림프구수에 영향을 미치는 것이 밝혀졌다. 그러나 면역기능을 평가하기 위해서는 림프구의 수 뿐만 아니라 기능에 대해서도 면밀히 검토해야 한다. 금후에는 운동처리구가 림프구의 기능에 미치는 영향을 해석할 필요성이 사료된다.

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Investigations for hematologic values and the differential count of white blood cell count (WBC) for Korean indigenous cattle (KIC) and endangered indigenous cattle (EIC) are rarely performed. Therefore, the objective of this study was to investigate the hematologic values of total 40 EIC (White, Black, Mini cattle) and 35 KIC as control by analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of EIC were significantly decreased by age (p<0.05). The mean values of RBC, HCT, MCV and MCHC between EIC and KIC of the same age (2 ～3 years) showed the statistical significance (p<0.05). Also, in the WBC of EIC, the mean values were decreased according to the age from 13.9×103/μL～12.7×103/μL under 1 year to 9.1×103/μL～11.5×103/μL over 2 years respectively. In the differential count of WBC of EIC (White, Black, Mini cattle), it showed generally the rates of 40.2%, 52.2%, 49.0% lymphocyte and 27.2%, 33.9%, 32.0% segmented neutrophil from 2～3 years respectively. Result of this study will be used for establishing reference range for blood analysis in EIC such as white, black and mini cattle. This study reported hematological values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of endangered Korean native cattle. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

The cloning efficiency is extremely low despite successful somatic cell nuclear transfer (SCNT) method producing cloned animals in several mammals. In general, faulty epigenetic modifications underlying the incomplete reprogramming of donor cell nuclei after SCNT mainly results in low cloning efficiency. The nuclear reprogramming process involves epigenetic modifications, such as DNA demethylation and histone acetylation, which may be an important factor in enhancing the cloning efficiency. Recently, the histone deacetylase inhibitors (HDACi), such as trichosatin A (TSA) and m-carboxycinnamic acid bishydroxamide (CBHA), to increase histone acetylation have been used to improve the developmental competence of SCNT embryos. Therefore, we compared the effects of TSA with CBHA on the in vitro developmental competence and pluripotency-related gene expression (Oct4, Nanog and Sox2) in porcine cloned blastocysts and histone acetylation pattern (H3K9ac). The porcine cloned embryos were treated with a 50nM concentration of TSA and 100μM concentration of CBHA during the in vitro early culture (10h) after cell fusion and then were assessed to cleavage rate, development to the blastocyst stage and pluripotency-related gene expression in NT blastocyst also, level of histone acetylation in zygote, 2cell, 4cell stage. As results, Although NT, TSA and CBHA treated NT embryos were not different between all groups for cleavage rates, the developmental competence to the blastocyst stage was significantly increased in CBHA treated embryos (22.7%) compared to that of normal NT and TSA treated NT embryos (8.1% and 15.4%)(p<0.05). In addition, all of pluripotent transcription factors (Oct4, Nanog and Sox2) were expressed in the CBHA treated NT embryos, however, Sox2 and Oct4 were expressed in TSA treated NT embryos and expression pattern of CBHA treated NT embryos is particularly similar to that of IVF embryos. Also, CBHA treated NT embryos were increased in level of histone acetylation (H3K9ac) at the zygote, 2-cell, 4-cell stage compared to those of NT and TSA treated NT embryos. In conclusion, the treatment of CBHA as a histone deacetylase inhibitor significantly increased the developmental competence of porcine NT embryos and pluripotency-related gene expressions(Oct4, Nanog and Sox2) in NT blastocysts and level of histone acetylation (H3K9ac).

본 시험은 한국 재래계와 도입계의 육성기(0 ~ 20주령) 성장 능력을 비교분석하기 위하여 수행하였다. 공시계는 가축유전자원 관리기관에서 종란을 인수하여 부화시킨 암수 병아리 180 수를 이용하였으며, 한국 재래계 1 계통(A)과 도입계 2계통(B, C)으로 총 3 계통의 육성률, 육성단계별 체중, 증체량, 사료 요구율, 정강이 길이 등을 조사하였다. 육성 초기(0 ~ 4주) 육성률은, A와 C계통은 98.33%로 높게 나타났으나, B계통은 91.67% 로 낮게 나타났다. 육성기 전 주령(0 ~ 20주)육성률도 C계통이 98.33%로 높게 조사 되었고, B계통이 83.33%로 낮게 조사되었다. 육성기 전 주령의 체중은 A, B계통이 C계통에 비해 높게 조사되었다(p < 0.05). 한국 재래계통인 A계통은 발생 시에는 체중이 가장 낮게 조사되었지만, 20주령에는 1.64 kg으로 가장 무거운 것으로 나타났다(p < 0.05). 계통간 사료 요구율은 0 ~ 8주령에서 C계통이 가장 높았고(p<0.05), 12 ~ 16주령에서는 B계통이 가장 높았으며(p < 0.05), 0 ~ 20주령에서는 유의적 차이가 없었다(p > 0.05). 육성 단계별 정강이 길이를 살펴보면, 0 ~ 12주령까지 B계통에서 가장 높게 나타났고, 16 ~ 20주령에서는 B와 C계통에서 높게 나타났다(p < 0.05). 마지막으로 계통간 정강이 길이 증가량은 0 ~ 8주령까지 B계통에서 가장 높게 나타났고, 8 ~ 16주령까지는 C계통에서 가장 높게 나타났다(p<0.05). 이와 같이 한국 재래계의 육성기 능력이 도입계에 비하여 다소 낮게 조사되었지만, 한국 재래계 고유 특성을 유지하고 있었다. 이러한 한국 재래계의 고유 특성들은 한국 재래계 활용에 있어 기준 자료로 이용될 것으로 사료된다(색인어 : 한국 재래계, 도입계, 육성기, 체중, 정강이 길이).

Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we investigated the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of apoptosis-related genes was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts (10.2x1014/mol s-1 versus 6.4x1014/mol s-1, p＜0.05). Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0 and over 12.0x1014/mol s-1, respectively. Apoptosis regulatory genes, Hsp-70.1 were significantly increased in over-10.0 group than below 10.0 group but in Caspase-3, Bax and P53 gene, there was no significant difference. In conclusion, These results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

This study was conducted for SNPs in the 5'-regions of estrogen receptor-α(ESR-α), and association with calving interval (CI), service per conception (SPC) and 305 days milk yield in Hanwoo and Holstein dairy cattle. The genet-ic improvement was incurred low reproduction performance. The objective of this study was to investigate connec-tions between single nucleotide polymorphisms (SNP) of Estrogen receptor-α (ESR-α) with reproduction performance (calving interval, service per conception, and 305 d milk yield) in Hanwoo and Holstein dairy cattle. Hanwoo and Holstein blood samples were collected from 183 and 124 dam of breeding farms and DNA was extracted. Primer design was based on NCBI GenBank (Accession No. AY340579). The PCR-RFLP method with Bgl I was used to ge-notype the cattle. The result showed two variants of the ESR-α gene. The Bgl I cut the 492 bp amplification pro- duct into 322 bp and 170 bp fragments for allele G, while allele A remained uncut, resulting in two restriction frag-ments for homozygote G/G and three fragments for heterozygote A/G. We found two of different genotypes in the-se breeds, A/G and G/G. In Hanwoo, the A/G genotype frequency was 0.13, and G/G was 0.87. The CI of A/G was 382.18±10.03 days, and G/G was 381.69±5.22 days. The SPC of A/G was 1.62±0.16, and G/G was 1.32±0.04. While CI showed no significance difference, SPC exhibited significant difference (p<0.05). In Holstein cattle, the frequency of genotype A/G was 0.02 and G/G was 0.98. The 305 days milk yield of A/G was 7,253.00±936.00 kg and of G/G was 8,747.51±204.88 kg, showing no significant difference.

본 연구의 목적은 염소의 개체식별에 있어서 효율적인 microsatellite (MS) marker set를 확립하는데 있다. 염소 10집단 455두를 대상으로 국제식량농업기구(FAO) 권고 및 미국 국립생물정보센터(NCBI)에서 수집한 30개 MS 마커에 대한 유전정보량을 확인한 후 대립유전자형 크기, 형광표지 종류, PCR 조건 등 을 고려하여 7개의 MS 마커(SRCRSP1, SRCRSP8, OarFCB048, OarFCB193, BOBT24, INRA063, DRBP1)를 선정하였다. 또한 분석 효율성을 극대화하기 위해 7개 마커에 대해서 multiplex PCR 기법을 적용하였다. 분석된 7개 MS 좌위에서 총 80개의 대립유전자형이 관찰되었고, 기대이형접합도(HExp) 및 다형정보지수(PIC)의 평균치는 0.710 및 0.674로 산출되었다. 7개 좌위의 다형성을 기반으로 무작위 교배집단, 반형매 교배집단 그리고 전형매 교배집단으로 가정하였을 때의 동일개체 출현확률은 각각 2.42×10-11, 1.70×10-8 및 2.22×10-4으로 산출되었다. 이상의 결과는 국내에서 사육되고 있는 염소의 개체수를 고려할 경우 동일개체 출현 가능성이 없는 것으로 판단된다. 따라서 본 연구에 의해서 구축된 염소 개체식별 마커 세트 및 분석방법은 우리나라 염소의 개량 및 안전유통에 유용하게 활용될 수 있을 것으로 사료된다.

Oxygen consumption is a useful parameter for evaluating mammalian embryo quality, since individual bovine embryos was noninvasively quantified by scanning electrochemical microscopy (SECM). Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we measured to investigate the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of pluripotent gene and anti-oxidant enzyme was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). Oxygen consumption of blastocyst was measured using a SECM and total cell number of in vitro blastocyst was enumerated by counting cells stained by propidium iodide. The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts (10.2 × 1015/mols—1 versus 6.4 × 1015/mols—1, p<0.05). Total cell numbers of in vitro blastocysts were 74.8, 90.7 and 110.2 in the oxygen consumption of below 10.0, 10.0∼12.0 and over 12.0∼1015/mols—1, respectively. Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0∼12.0 and over 12.0 × 1015/mols—1, respectively. GPX1 and SOD1 were significantly increased in over —10.0 group than below 10.0 groups but in catalase gene, there was no significant difference. On the other hand, In OCT-4 and Sox2, pluripotent gene, there was a significant difference (p<0.05) between the below-10.0 (0.98 ± 0.1) and over 10.0 (1.79 ± 0.2). In conclusion, these results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

핵이식(NT) 기술을 이용하여 여러 동물 종에서 성공적으로 복제산자가 보고되고 있지만, 아직까지 비효율적인 기술로 남아있다. 본 연구에서는 돼지 체세포 복제 생산 효율성을 증진시키기 위한 방안으로 수핵난자의 품질에 초점을 맞추어 Brilliant cresyl blue (BCB) 염색을 통하여 발육능이 우수한 미성숙 난자를 선발하고, 난자의 감수분열 재개에 관여하는 단백질 합성을 비특이적으로 억제하는 cycloheximide (CHXM)을 이용하여 돼지 난자의 감수분열 재개를 억제시켜 난자의 성숙 동기화를 유도하였다. 또한 핵초기화에 밀접한 영향을 주는 핵막붕괴(NEBD)와 조기염색체응축 (PCC)을 유도하는 MPF의 활성화를 높이기 위하여 단백질 phosphatase 억제제인 caffeine을 첨가하여 수핵난자의 품질을 향상시키고자 하였다. 실험 방법으로는 13 mM BCB 첨가된 배양액에 90분 동안 미성숙난자를 배양하여 BCB 용액의 착색 여부를 구분하여 선발하고, 5 ㎍/ml CHXM를 체외 성숙액에 첨가하여 난자성숙 동기화를 유도하였다. 또한 탈핵 후 탈핵난자를 caffeine을 처리하여 세포주기 관련 단백질의 활성화를 인위적으로 조절하여 체세포복제 수핵난자로 사용하였다. 실험 결과로서 BCB 염색 돼지 미성숙 난자를 대조구와 비교할 때 제2 감수분열 중기(MII)에 도달하는 체외성숙율과 단위발생란의 배반포기까지의 체외 발육율이 유의적으로 증가하는 것이 관찰되었다. 또한 미성숙 돼지 난자의 초기 성숙 (12∼16시간)에 CHXM를 처리하였더니 난자 감수분열 재개가 억제되어 GV기에 핵 성숙이 정지되어 동기화가 유도되었다. GV기에 세포주기 동기화된 난자들은 CHXM를 제거하였을 때 난자 성숙의 진행속도도 일치하는 것이 관찰되었다. 이런 결과는 가장 적합한 탈핵시기인 제1 감수분열 후/말기(AI/TI)에 난자들이 다수 분포하여 대조구에 비하여 높은 탈핵율 (87.9%)을 얻을 수 있었다 (P < 0.05). 덧붙여 5 mM의 caffeine을 돼지 난자에 12시간 처리하였을 때 난자 MPF의 활성화가 증가하는 것이 관찰되었지만 (P < 0.05), 10 mM caffeine 농도를 처리하였을 때 MPF의 활성화가 오히려 감소되어 단위발생란의 배반포기까지의 체외발육에도 악영향을 주는 것이 관찰되었다.

The aim of this study was to identify allele variability and frequencies of six microsatellite markers (BM861, INRA124, INRA189, UMN0103, UMN0307 and UMN0504) specific to the Y-chromosome. DNA samples from 147 males of three Korean native and seven exotic cattle breeds were tested. Three (Chikso, Heugu, and Jeju black cattle) Korean native cattle (KNC) breeds showed Bos taurus genotype. In the neighbor-joining tree, based on Nei’s DA genetic distance, ten breeds represented main group. In addition, Bayesian clustering result showed that 3 main clusters are for individual allele variability and frequencies. Moreover, KNC breeds showed differences in genotype with B. taurus type when compared to previous studies. The molecular information of paternal lineage in this study would be useful for the conservation and utilization of three KNC breeds as genetic resources.

This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of 2×108/ml by doubling in every 30 minutes at 4℃ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to LN2. The presented straws were examined the viability and motility after thawed at 37℃ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm (93.27±1.62%) than frozen-thawed sperm (73.34±3.27%). However, there were no significant differences between fresh and frozen-thawed dead cell rate (7.35±2.63 vs, 13.71±2.85). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different (72.86±2.83 vs, 81.47±2.48), similarly, the dead cell rates was similar (18.41±3.42% and 17.26±4.25). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

The study was conducted to investigate the comparison of pregnancy rate and transferable embryos produced by genetically superior Korean cows (Hanwoo) of livestock farms. Eighteen Hanwoo donors were superovulated with gonadotropin for 4 days combined with Progesterone Releasing Intravaginal. Embryos were recovered 7 days after the second insemination by flushing the uterus with embryo collection medium. No differences were observed in the efficiency of rate of superovulation in groups A (low nutrition) and B (highnutrition) it was observed to be 100.0% and 87.5%, respectively. The mean numbers of total embryos were 10.8±3.4 and 8.9±2.5, and transferable embryos were 7.5±3.3 and 4.0±1.5 in groups A and B, respectively. The pregnancy rates after embryo transfer were 23.5%, 20.0%, C 80.0% and 55.6% in farm A, B, C, and D, respectively. In conclusion, results suggest that superovulation could be used quite effectively to raise superior Hanwooembryos. However, physical and biological condition of recipients greatly affects the rate of pregnancy.