한우, 칡소 및 제주 흑우 Calpain-Calpastatin 유전자 다양성

이승환,김승창,조수현,최봉환,Aditi Sharma,임다정,당창권,장선식,김재환,고문석,양보석,강희설 충남대학교 농업과학연구소 2013 농업과학연구 Vol.40 No.2

The aim of study was to investigate genetic diversity for the calpain/calpastatin gene in three Hanwoo breeds [(Brown (n=62), Brindle (n=81) and Jeju Black (n=30)]. Random samples from three breeds of Hanwoo were selected and genotyped for the 7 SNPs of calpain/calpastatin using TaqMan method. Allele frequencies were investigated for CAPN1/CAST gene. Allele frequency of CAST2 SNP was 0.75, 0.59 and 0.22 for Brown, Brindle and Jeju black, respectively. The CAST3 revealed allele frequency of 0.59 and 0.57 in Brown and Jeju Black, while it showed very low allele frequency (0.07) in Brindle. In particular, favorable allele (G allele) for the CAPN1-2 SNP which was shown a strong association with tenderness in Taurine and Indicine cattle revealed 16% and 17% higher allele frequency in Brown Hanwoo (0.82) comparing Brindle (0.66) and Jeju Black Hanwoo (0.65). AMOVA demonstrated that among population variance occupied only 10% of total variance and among individual variance was 0%, while within individual variance was 90% of total variance. This result showed that population effect contributed very small portion of genetic to these three Hanwoo breeds, while within individual variance contributed large portion of genetic diversity within these Hanwoo breeds. In conclusion, three Hanwoo breeds (Brown, Brindle and Jeju black) showed a genetically homogeneous based on the 7 SNPs of CAPN1/CAST gene and it came from same ancestor to form modern Hanwoo breed.

The effect of butylated hydroxytoluene (BHT) addition on sperm motility, viability, acrosomal integrity and plasma membrane integrity

Byoung-Chul Yang,Sung-Sik Kang,Ui-Hyung Kim,Hyun-Ju Kim,Boh-Suk Yang,Chung Ki-Yong,Jang Sun-Sik,Seok-Dong Lee,Myoung-Sook Lee,Sang-Rae Cho 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

The present study was conducted to investigate the effect of BHT supplementation on sperm motility, viability, acrosomal integrity and plasma membrane integrity after frozen-thawing. One ejaculate was collected from one fertile Hanwoo bull by using artificial vagina at Hanwoo Research Institute. The ejaculate was transferred to laboratory immediately and diluted with pre-warmed semen extender (Optixcell, France) (1:1). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and divided into 5 groups according to BHT concentration (0, 0.5, 1.0, 2.0 and 4.0 mM) and cryopreserved in LN2 tank until evaluation. Frozen-thawed semen was transferred to 1.5 ml tube and incubated for 0, 2 and 4h. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain). There were not significant effects of BHT supplementation (0, 0.5, 1.0 and 2.0 mM) on total motile and VSL with 25μm≥ at 0, 2 and 4h. However, 4.0 mM of BHT supplementation showed negative effect on total motile (26.3%), VSL with 25μm≥ (1.3%) at 0 h (p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method and divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). There were no significant differences of LIA, LDA, DIA and DDA on various BHT concentrations at 0 and 2 h. However, 4.0 mM BHT supplementation showed decreased LIA compared with 0, 0.5, 1.0 and 2.0 mM BHT at 4 h (34.6, 37.1, 43.6, 45.4 and 14.7% vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.01). Addition of 4.0 mM of BHT showed negative effect on plasma membrane integrity compared with that of 0, 0.5, 1.0 and 2.0 BHT at 2 h (71.9, 64.2, 64.6, 67.5 and 31.7 % vs. 0, 0.5, 1.0, 2.0 and 4.0 mM, irrespectively; p<0.05). In conclusion, various BHT concentrations on optixcell extender showed no improvement on sperm motility, viability and plasma membrane integrity.

Fragmentation and development of preimplantation porcine embryos derived by parthenogenetic activation and nuclear transfer

Im, Gi-Sun,Yang, Boh-Suk,Lai, Liangxue,Liu, Zhonghua,Hao, Yanhong,Prather, Randall S. JOHN WILEY & SONS LTD 2005 Molecular Reproduction and Development Vol.71 No.2

<P>Fragmentation occurs during early developmental stages of electrically activated oocytes and nuclear transfer (NT) embryos. It might contribute to the low developmental rate of porcine NT embryos. The present study was conducted to investigate whether the addition of sugars such as sorbitol or sucrose suppresses fragmentation and supports the development of electrically activated oocytes and NT embryos. The activated oocytes were cultured in Porcine Zygote Medium-3 (PZM-3) supplemented with sorbitol or sucrose for 2 days after electric activation, and then cultured in the PZM-3 for the remaining 4 days. The osmolarities of PZM-3, PZM-3 supplemented with 0.05 or 0.1 M sorbitol, and PZM-3 with 0.05 M sucrose were 269 ± 6.31, 316 ± 3.13, 362 ± 4.37, and 315 ± 5.03 mOsm, respectively. When parthenogentically activated oocytes were cultured in PZM-3 supplemented with 0.05 M sorbitol or sucrose for the first 2 days and then cultured in PZM-3 without sugar, a significantly higher (P < 0.05) cleavage rate and blastocyst rate were observed. Interestingly, addition of sugar to PZM-3 for 2 days reduced the fragmentation rate compared to PZM-3 without sugar. In NT embryos, sugar addition into PZM-3 increased the fusion rate (84.2% ± 6.07 vs. 95.1% ± 2.52), cleavage rate (67.6% ± 5.80 vs. 77.3% ± 3.03), and developmental rate to the blastocyst stage (10.2% ± 0.79 vs. 19.4% ± 1.77). There was no significant difference between treatments for the number of the blastocysts. In addition the fragmentation rate was reduced compared to PZM-3 without sorbitol (26.1 ± 4.30 vs. 14.5 ± 1.74). In conclusion, increasing the osmolarity of PZM-3 through addition of either sorbitol or sucrose for 48 hr increased the cleavage and developmental rate to the blastocyst stage by reducing the fragmentation rate through increasing osmolarity. Mol. Reprod. Dev. 71: 159–165, 2005. © 2005 Wiley-Liss, Inc.</P>

Perinatal Phthalate/Adipate Esters Exposure in Rats: Effects on Maternal Body Weight Changes and Developmental Landmarks in Offspring Rats

Lee, Hwi-Cheul,Yang, Byoung-Chul,Ko, Yeoung-Gyu,Kim, Dong-Hoon,Park, Jin-Ki,Yang, Boh-Suk,Seong, Hwan-Hoo,Yamanouchi, Keitaro,Nishihara, Masugi The Korean Society of Animal Reproduction 2006 Reproductive & developmental biology Vol.30 No.4

Di-n-butyl phthalate (DBP), diisononyl phthalate (DINP) and di-(2-ethylhexyl) adipate (DEHA) are ubiquitously distributed chemicals that are widely used as plasticizers and also found at low levels in foods. The aims of this study were to determine whether perinatal exposure to DBP, DINP and DEHA could alter normal patterns of neonatal development. Dams were provided with pulverized soy-free diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4.000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day 15 to postnatal day 21. Exposure to the high doses of DBP, DINP and DEHA during gestational period significantly decreased food consumption and body weight gain of dams. These chemicals reduced neonatal body weight as well as that of the after maturation. Also, exposure to DINP of all the doses used and the higher doses (2,400 and 12,000 ppm) of DEHA decreased AGD at PND 1 in male neonates, though that to DBP did not affect AGD in males. In female neonates, an increase in AGD was observed in DBP- and DINP-exposed animals at the highest doses. Moreover, these chemicals affected survival rate of pups at PND 5, and delayed onset of eye opening in all chemica1-exposed groups at PND 17. These results suggest that perinatal exposure to these chemicals may affect the normal development and/or growth of offspring.

The effect of different heights of liquid nitrogen (LN2) vapor on Hanwoo sperm motility and morphology after frozen-thawing

Sung-Sik Kang,Byoung-Chul Yang,Ui-Hyung Kim,Hyun-Ju Kim,Boh-Suk Yang,Ki-Yong Chung,Sun-Sik Jang,Seok-Dong Lee,Myoung-Sook Lee,Sang-Rae Cho 한국수정란이식학회 2017 한국수정란이식학회 학술대회 Vol.2017 No.05

The present study was conducted to investigate the effect of different heights from liquid nitrogen (LN2) vapor on sperm motility and morphology after frozen-thawing. Two ejaculates were collected from 2 fertile Hanwoo bulls (A and B) by using artificial vagina at Hanwoo Research Institute. After collection, ejaculates were transferred to laboratory immediately and diluted with semen extender (Optixcell, France). Sperm dilutions were extended to a final concentration of 40 x 106 sperm/ml, and cooled at 4°C for 4 h and loaded to 0.5 ml straws. The straws were divided into 2 groups. Straws were placed in 3 or 9 cm of LN2 vapor for 14 min and then plunged into LN2 tank and cryopreserved until evaluation. Sperm motility and motility parameters (total motility, VSL with 25μm≥, VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were evaluated by sperm class analysis (SCA, IVOS, Spain) after frozen-thawed. In bull A, 3cm group showed higher percentages of total motility, VSL with 25μm and VAP compared those with 9cm group (98.0 vs. 93.4%, 62.4 vs. 54.0% and 98.6 vs. 93.2%, 3 vs. 9 cm, irrespectively; p<0.001). In bull B, frozen-thawed sperm of 3cm group showed higher percentages of VSL with 25μm, VCL, VSL, VAP and BCF compared with those of 9cm group (43.5 vs. 26.0%, 123.8 vs. 111.6 μm, 62.9 vs. 57.3 μm and 81.5 vs. 72.5 μm; 3 vs. 9 cm, irrespectively; p<0.001). The viability and acrosomal integrity of spermatozoa were evaluated by Trypanblue/Giemsa staining method divided into 4 groups; live and intact acrosome (LIA), live and damaged acrosome (LDA), dead intact acrosome integrity (DIA), dead damaged acrosome (DDA). In bull A, frozen-thawed sperm of 3 and 9cm groups showed no significant difference in LIA, LDA, DIA and DDA. In bull B, 3 cm group showed higher LIA and lower DIA compared with those of 9 cm group (73.2 vs. 23.7% and 23.7 vs. 32.2%, 3 vs. 9 cm, irrespectively; p<0.001). We suspected that 3 cm vapor on LN2 vapor might be affected positively spermatozoa viability and acrosomal integrity compared with 9 cm group. In conclusion, semen freezing procedure in the present study will improve sperm quality after frozen thawing.

종돈개량 모니터링시스템에 대한 고찰

도창희(Chang-Hee Do),양창범(Chang-Beom Yang),최재관(Jae-Gwan Choi),양보석(Boh-Suk Yang),송형준(Hyung-Jun Song) 충남대학교 농업과학연구소 2015 농업과학연구 Vol.42 No.3

This paper sketches the strategies and designs for monitoring system of swine genetic improvement. The system should reflect every side of pig production. The system leads us to assess the efficiency of pig production and the scope of the system includes not only nucleus, multiplying and commercial herds, but also packing and processing sectors. For more accurate statistics, data for this monitoring system must be collected from all above mentioned areas, but not by random sampling. Futhermore, data analysis results including seedstocks and distribution information of genetic trend should be included in the system. The schema of knowledge database system could be employed in the system. The monitoring system in the final destination would unify the systems derived from various sources and provide any solution in swine industry including pig breeding.

Perinatal PhthalateI/Adipate Esters Exposure in Rats: Effects on Maternal Body Weight Changes and Developmental Landmarks in Offspring Rats

Hwi-Cheul Lee,Byoung-Chul Yang,Yeoung-Gyu KO,Dong-Hoon Kim,Jin-Ki Park,Boh-Suk Yang,Hwan-Hoo Seong,Keitaro Yamanouchi,Masugi Nishihara 한국동물생명공학회(구 한국동물번식학회) 2006 Reproductive & developmental biology Vol.30 No.4

Di-n-butyl phthalate (DBP), diisononyl phthalate (DINP) and di-(2-ethylhexyl) adipate (DEHA) are ubiquitously distributed chemicals that are widely used as plasticizers and also found at low levels in foods. The aims of this study were to determine whether perinatal exposure to DBP, DINP and DEHA could alter normal patterns of neonatal development. Dams were provided with pulverized soy-free diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day 15 to postnatal day 21. Exposure to the high doses of DBP, DINP and DEHA during gestational period significantly decreased food consumption and body weight gain of dams. These chemicals reduced neonatal body weight as well as that of the after maturation. Also, exposure to DINP of all the doses used and the higher doses (2,400 and 12,000 ppm) of DEHA decreased AGD at PND 1 in male neonates, though that to DBP did not affect AGD in males. In female neonates, an increase in AGD was observed in DBP- and DINP-exposed animals at the highest doses. Moreover, these chemicals affected survival rate of pups at PND 5, and delayed onset of eye opening in all chemical-exposed groups at PND 17. These results suggest that perinatal exposure to these chemicals may affect the normal development and / or growth of offspring.

Perinatal Phthalate/Adipate Esters Exposure in Rats: Effects on Maternal Body Weight Changes and Developmental Landmarks in Offspring Rats

Hwi-Cheul Lee,Byoung-Chul Yang,Yeoung-Gyu Ko,김동훈,Jin-Ki Park,Boh-Suk Yang,Hwan-Hoo Seong,Keitaro Yamanouchi,Masugi Nishihara 사단법인 한국동물생명공학회 2006 Reproductive & developmental biology Vol.30 No.4

Di-n-butyl phthalate (DBP), diisononyl phthalate (DINP) and di-(2-ethylhexyl) adipate (DEHA) are ubiquitously distributed chemicals that are widely used as plasticizers and also found at low levels in foods. The aims of this study were to determine whether perinatal exposure to DBP, DINP and DEHA could alter normal patterns of neonatal development. Dams were provided with pulverized soy-free diet containing 20, 200, 2,000 and 10,000 ppm of DBP, 40, 400, 4,000 and 20,000 ppm of DINP, or 480, 2,400 and 12,000 ppm of DEHA from gestational day 15 to postnatal day 21. Exposure to the high doses of DBP, DINP and DEHA during gestational period significantly decreased food consumption and body weight gain of dams. These chemicals reduced neonatal body weight as well as that of the after maturation. Also, exposure to DINP of all the doses used and the higher doses (2,400 and 12,000 ppm) of DEHA decreased AGD at PND 1 in male neonates, though that to DBP did not affect AGD in males. In female neonates, an increase in AGD was observed in DBP- and DINP-exposed animals at the highest doses. Moreover, these chemicals affected survival rate of pups at PND 5, and delayed onset of eye opening in all chemical-exposed groups at PND 17. These results suggest that perinatal exposure to these chemicals may affect the normal development and / or growth of offspring.

Verification of Enuc1eation Time in In Vitro Matured Bovine Oocytes by Meiotic Spindle Imaging with the PolScope

SeWoong Kim,Min-Jung Lee,Gi-Sun Im,Byoung-Chul Yang,Hwan-Hoo Seong,Boh-Suk Yang,Hee-Tae Cheong,Dong-Hoon Kim 한국동물생명공학회(구 한국동물번식학회) 2007 Reproductive & developmental biology Vol.31 No.2

Melanocyte-stimulating Hormone Receptor (MC1R) Genotype and Its Effects on Coat Color in Korean Jindo Dogs

Hong, Kyung-Won,Kim, Sang-Wook,Jang, Hong-Chul,Yang, Seung-Min,Shin, Young-Bin,Hong, Yoon-Hye,Kim, Jong-Seok,Oh, Seok-Il,Choi, Yoon-Ju,Chung, Dong-Hee,Yang, Boh-Suk,Lee, Ji-Woong,Choi, Bong-Hwan Asian Australasian Association of Animal Productio 2009 Animal Bioscience Vol.22 No.8

The Jindo dog is a Korean natural monument and is recognized by the Fédération Cynologique Internationale. A prominent feature is the diverse coat color within the breed. To analyze the genetic basis of variation in the Jindo coat color, we sequenced the protein-coding regions of the melanocortin 1 receptor gene (MC1R). The MC1R coding sequence was determined from 154 dogs in five breeds (Jindo, Labrador Retriever, English Springer Spaniel, Belgian Malinois, and German Shepherd). To confirm the genetic structure of sampled populations, we tested for Hardy-Weinberg equilibrium (HWE) and computed $F_{st}$ The sample populations did not significantly deviate from HWE. $F_{st}$ was 0.02 between white and fawn Jindo dogs; this was lower than $F_{st}$ between breeds. Six single nucleotide polymorphisms (SNPs) were detected in the MC1R coding region. Among the six SNPs, five were non-synonymous (S90G, T105A, Q159P, M264V, and R306ter) and one was synonymous SNP (Y298Y). From the SNPs, we predicted four haplotypes (H1, H2, H3, and H4) for Jindo MC1R. Jindo dogs had different haplotypes corresponding to different coat colors. H1 was frequently observed in white Jindo dogs with an odds ratio of 5.03 (95% CI: 2.27-11.18, p<0.0001), whereas H2 and H4 were observed only in fawn Jindo dogs. Our findings indicate that SNP haplotype can influence coat color. Knowledge of MC1R haplotypes can help discriminate white and fawn coats in Jindo dogs. We hope this report will trigger more research into the genetics of this traditional Korean dog and will be a reference for dogs of Asian origin. Also, our results will provide a useful genetic marker for Jindo dog breeders who have selected for specific colors.