The purpose of this study is to expression pattern of melanogenesis associate genes on cultured melanocyte layer cells in Korean Brindle Cattle(Dark, Brindle and Yellow) were analyzed to evaluate the effects of sex hormones on the control of melanogenesis pathways. Korean Brindle Cattle(Dark, Brindle and Yellow) melanocyte in the skin cells was collected. after the addition of estrogen and testosterone, the culture was analyzed for expression of cell activity and melanin genes for 72 hours. For the analysis of estrogen in different coat color other than the melanogenesis-related genes it is increasingly yellow showed low expression. in particular, the cells of the brindle coat color is low active and expression of genes. However, the testosterone was low, the expression of cell activity inhibiting MMP-2. the expression of melanin genes actually showed a tendency to increase gradually, which is testosterone compared with the estrogen to be considered that affect the skin cell layer brindle coat color. In this study, stimulation with estrogen triggered the inhibition of MC1R of the melanocyte in brindle coat color, but testosterone is induced MC1R in melanocyte. Therefore, considered the eumelanin or phaeomelanin activation are controlled caused by differential expression of sex hormones on melanocyte in Korean Brindle Cattle.

The coat color of mammals is determined by the melanogenesis pathway, which is responsible for maintaining the balance between black-brown eumelanin and yellow-reddish pheomelanin. The melanogenesis-associated genes controlling pigmentation act as a complex and interact with each other to cause phenotypic and genotypic variations in cattle. That the MC1R genotype of Korean native cattle with dark muzzle was e/e or E+/e, while the genotype of Korean native cattle with light muzzle was E+/E+, which is a variant of the MC1R genotype in the Korean native cattle. Especially, the MC1R expression type is shows how much pigmentation, important factor in deciding its status in the coat and nose colours. However, information regarding the coat or nose colours-associated gene regulation of korean cattle is not yet unknown. Therefore, in this study was to investigate the expression patterns of melanogenesis-associated genes in black dot nose(korea brindle cattle) and normal nose(korea native cattle). Using microarray clustering and real-time polymerase chain reaction techniques, we analysed that the expression of genes involved in the mitogen-activated protein kinase (MAPK) and Wnt signaling pathways is distinctively regulated in the dark and light muzzle tissues. Differential expression of tyrosinase was also noticed, although the difference was not as distinct as those of MAPK and Wnt. We hypothesize that emphasis on the MAPK pathway in the Korea brindle cattle induces eumelanin synthesis through the activation of cAMP response elementbinding protein and tyrosinase, while activation of Wnt signaling counteracts this process and raises the amount of pheomelanin in the native cattle. Regarding the increasing interest in the genetic diversity of cattle stocks, genes we identified for differential expression in the brindle cattle vs. native cattle may serve as novel markers for genetic diversity among cows based on the coat and muzzle color phenotype.

This study was conducted to report the census survey results of south Korean Chikso (Korean Brindle Cattle) distribution and to diagnose the current coat color expression patterns. Two years Chikso census showed that there were 2,413 Chikso in 2013 and 2,754 in 2014 in south Korea. Number of animals between 0 to 35 months of age was 1,632 heads in 2014 which comprised 59% of total Chikso population size. The percentages of animals between 36 ~ 71 months of age and over 6 years of age were 31% and 10% of total. Out of 2,416 animals with class numbers assessed by their coat colors, 1,249 heads (52%) were in group I (with black stripes, class no. 1 ~ 3) and the others (class no. 4 ~ 7) were in group II (1,167 heads, 48%). Among the 1,551 animals that were photographed twice in 2013 and in 2014, 226 animals were assessed different coat color class numbers, and round 90% of which were within the ages less than 48 months. The number of animals switched in coat color pattern groups over a year was 56, which was 3.6% of total number of animals on survey. And around 88% of the animals switched in group category was of the animals younger than 24 months of age. Therefore, we conclude that the coat color pattern becomes rather stable at around 24 months of age in Chikso populaion in south Korea.

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.

The aim of the present study was to identify coat color associated genes that are differentially expressed in mature Korean brindle cattle (KBC) with different coat colors and in Hanwoo cows. KBC calves, before and after coat color appearance, were included. Total cellular RNA was isolated from the tail hair cells and used for microarray. The number of expressed coat color associated genes/probes was 5813 in mature KBC and Hanwoo cows. Among the expressed coat color associated genes/probes, 167 genes were the coat color associated genes listed in the Gene card database and 125 genes were the pigment and melanocyte genes listed in the Gene ontology_bovine database. There were 23 genes/probes commonly listed in both databases and their expressions were further studied. Out of the 23 genes/probes, MLPH, PMEL, TYR and TYRP1 genes were expressed at least two fold higher (p<0.01) levels in KBC with brindle color than either Hanwoo or KBC with brown color. TYRP1 expression was 22.96 or 19.89 fold higher (p<0.01) in KBC with brindle color than either Hanwoo or KBC with brown color, respectively, which was the biggest fold difference. The hierarchical clustering analysis indicated that MLPH, PMEL, TYR and TYRP1 were the highly expressed genes in mature cattle. There were only a few genes differentially expressed after coat color appearance in KBC calves. Studies on the regulation and mechanism of gene expression of highly expressed genes would be next steps to better understand coat color determination and to improve brindle coat color appearance in KBC.

The aim of this study was to determine the relationship between the coat color appearance of Korean brindle cattle and the changes of relevant hormone levels that may affect the hair pigmentation during different stages of growth and maturation. In mature cattle, levels of both ACTH and DHEA in Korean brindle cattle with brown color were significantly 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 at 2 to 6 months, the concentration of estradiol was significantly higher in calves with whole brindle color than those with part brindle color (p<0.05), when the coat color was confirmed. After 6 month of coat color confirmation, levels of testosterone and ACTH increased in calves with part brindle color and were significantly higher than those with whole brindle color (p<0.05). In calves of Korean brindle cattle at 1 or 2 months, there were no significant differences in 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 and maturation may affect the pigmentation of coat during the development of cattle. In addition to the current study correlating the different coat colors with relevant hormone levels, investigation of the coat color associated genes expressed in Korean brindle cattle may further clarify the mechanisms of coat color changes during their development.

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

The objective of this study was to determine the effect of the MC1R genotypes of the Chikso (Korean brindle cattle) sires on the coat colors of their offspring. In this study, 15 Chikso sires with known MC1R genotypes were used for breeding in the Gangwon Province Livestock Research Center, the Chungbuk Institute of Livestock and Veterinary Research, and the Livestock Experiment Station, Jeonbuk Institute of Livestock and Veterinary Research from either 2011 or 2012 to 2013. There were 6 sires with E+E+ genotypes and 9 sires with E+e genotypes, and their coat colors were all whole brindle (more than 50 of the body). Among the 90 calves produced in 2011∼2013 or 2012∼2013 from the 15 sires, 50 (55.6%) of them were females and 40 (44.4%) of them were males. Coat colors of the offspring were determined when they reached over 6 months of age. Calves with whole brindle, part brindle, brown and black coat colors were 42 (48.3%), 11 (12.6%), 18 (20.7%) and 16 (18.4%), respectively. Ratio of calves with whole brindle coat color was higher than any other coat colors. Among the offspring with whole brindle color, 20 (41.7%) calves were female and 22 (51.3%) calves were male. By determining the MC1R genotypes of the dams and calves in this study along the family lines, and investigating other genes that may be involved in the coat colors of the Chikso, better breeding system may be established to increase the brindle coat color appearance in the future.

Bovine coat color is decided by the melanocortin receptor 1 (MC1R) genotype mutation and melanogenesis. Specially, in the various cattle breeds, dominant black coat color is expressed by dominant genotype of ED, red or brown is expressed in the frame shift mutation of recessive homozygous e by base pair deletion and wild type of E+ is expressed in various coat colors. However, not very well known about the effected of MC1R genotype mutation on the coat color through family lines in KBC. Therefore, this study were to investigate effect of MC1R genotype mutation on the coat color, and to suggest mating breed system in accordance with of MC1R genotype for increased on brindle coat color appearance. Parents (sire 2 heads and dam 3 heads) and offspring (total : 54 heads) from crossbreeding in KBC family line with the MC1R genotype and phenotype records were selected as experimental animals. The relationship between melanocortin 1 receptor (MC1R) genotypes expression verified by PCR-RFLP, and brindle coat color appearance to the family line of the cross mating breed from MC1R genotype pattern was determined. As a result, 4MC1R genetic variations, E+/E+ (sire 1), E+/e (sire 2 and dam 3), E+/e with 4 bands of 174, 207 and 328 bp (dam 1) and E+/e with 3 bands of 174, 207, 328 and 535 bp (dam 2) from parents (sire and dam) of KBC. However, 3 genetic variations, e/e (24%), E+/E+ (22%) and E+/e (56%) were identified in offspring. Also, brindle coat color expressrated was the e/e with the 0%, E+/E+ with 67% and E+/e with 77% from MC1R genotype in offspring on the cross mating of KBC. Furthermore, when the sire had E+/e genotype and the dam had E+/E+ with the 3 bands or E+/e genotype, and both had whole body-brindle coat color, 62% of the offspring had whole body-brindle coat color. Therefore, the seresults, the mating system from MC1R genotype patterns of the sires (E+/e) and dams (E+/E+ with the 3 bands or E+/e) with brindle coat color may have the highest whole body-brindle coat color expression in their offspring.

The role of Fatty acid binding protein 4 (FABP4) gene is critical for lipid metabolism and for maintaining homeostasis in adipocytes. Association between Hanwoo carcass traits and FABP4 gene g.7516G>C SNP has been reported previously, however, its association and how does it influence Korean brindle and black cattle has not been demonstrated and established till date. For this purpose, the study was planned to analyze the SNP association (Single Nucleotide Polymorphism) (g.7516G>C) in FABP4 gene and gather genetic information on economic traits of Korean brindle cattle and black cattle. As per sequence of bovine FABP4 gene available (Genbank accession No. NC_007312.4), one pair of primers (5’-ATA TAG TCC ATA GGG TGG CAA AGA-3’ and 5’-AAC CTC TCT TTG AAT TCT CCA TTC T-3’) was designed to amplify a 452bp product of the FABP4 gene including the region of 7417–7868. The SNP, detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) methods using restriction enzyme MspA1I, was genotyped in 117 animals of brindle cattle and 24 animals of black cattle population. Statistical analysis revealed that the FABP4 genotype significantly (p<0.05) affect with carcass weight, but there was no significant association with any other economic traits was observed in brindle as well as in black cattle. In conclusion, these results suggested that SNP (g.7516G>C), located in FABP4 gene, could be used as important DNA marker of economic traits in Korean brindle cattle. Furthermore, we suggest that additional samples needs to further analysed to make related data exclusively authentic.

The melanocortin receptor type 4 (MC4R) gene is expressed in the hypothalamus and regulates energy intake and body weight. Recently, it has been reported that obesity and energy balance in human were also regulated by the MC4R gene. Therefore the objective of this study was to identify the polymorphism on the MC4R gene SNP C1786T and its association with economic traits in Korean native cattle (brindle and black cattle) by PCR-RFLP. A total of 125 cattle from the two breeds were tested for economic traits (meat quality index, backfat, thickness, carcass weight, longissimus muscle area and marbling score) and data was analyzed using SAS program. In the results, C allele had highest frequency than G allele frequency in the both breeds and the gene was significantly associated with meat quantity index and backfat thickness in brindle cattle breed. However, in black cattle, the gene was significantly associated with longissimus muscle area (p<0.05). These results suggest that C1786T SNP of the MC4R gene may be useful as a genetic marker for economic traits in the brindle and black cattle.

After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q -ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.

There are four (yellow, brindle, black, black in Jeju) kinds of native cattle in Korea. But only twelve hundred Korean brindle cattle (KBC) are remaining in limited areas of Korea and the genetic lineage, diversity, polymorphisms of KBC has not been identified. To analysis genetic polymorphism of KBC, 33 KBC were characterized using 11 microsatellite markers. Size of microsatellite marker was decided using Gene Mapper software after analysis ABI 3130XL. The average of allele numbers of KBC was 6.7 in this study, but that of Hanwoo was 10.0 in our previous report. The observed and expected heterozygosities of KBC were 0.719 and 0.738 but those of Hanwoo were 0.751 and 0.760 in our previous report. Also polymorphism information content (PIC) values were average 0.690 in KBC but 0.725 in Hanwoo. These results demonstrate that genetic polymorphism of KBC have decreased because the population was limited.

칡소의 귀세포를 이용한 핵이식 시에 전기 융합조건이 핵이식란의 발생에 미치는 영향을 알아보고, 칡소의 귀세포와 한우의 태아섬유아세포를 이용한 핵이식란을 이식하여 수태율을 비교하였다. 전기자극 조건에 따른 핵이식 후 융합율은 51∼68%의 범위로 차이는 없었으나, lysis 율은 1.9kv/cm의 10us, 20us에서 각각 0.0과 38.7%, 2.0kv/cm의 10us, 20us에서 각각 2.9와 37.5%, 2.1kv/cm의 10us, 20us에서 각각 21.2와 51.8%을 보였다. 난할율은 1.9kv/cm의 10us, 20us에서 각각 75.8과 69.8%, 2.0kv/cm의 10us, 20us에서 각각 76.9와 68.8%, 2.1kv/cm의 10us, 20us에서 각각 70.5와 68.5로 큰 차이가 없었다. 그러나 배반포 발생율은 1.9kv/cm의 10us, 20us 각각 19.5와 48.6%, 2.0kv/cm의 10us, 20us에서 각각 20.0과 40.9%, 2.1kv/cm의 10us, 20us에서 각각 44.2와 27.0%로 각 조건에서 시간에 따른 차이를 보였다. 핵이식에서 공여세포와 세포질간의 융합율과 lysis율은 전압과 통전시간에 영향을 받으며 전압을 높이는 것이 통전시간을 길게 하는 경우보다 수핵난자의 세포질에 상해를 줄이고 배반포 발생에 유리 하였다. 한우의 태아섬유아세포의 핵이식후 생산된 배반포를 5마리의 수란우에 이식한 결과 두 마리가 임신되었고(40.0%), 칡소의 귀 세포를 핵이식한 후 생산된 배반포를 19마리에 이식한 결과 3마리가 임신되었다(15.8%).

본 연구는 수정란 이식에 의해 생산된 울릉칡소의 모색분포의 조사를 통해 울릉칡소의 모색 고정에 필요한 기초자료로 활용하고자 실시하였다. 1. 울릉칡소의 모색 분포는 황모 24.3%(67/276두), 흑모 13.0%(36/276두)와 호반모 62.7% (173/276두)의 분포를 보였으며, 입식우의 호반모 출현율이 66%(105/159두)로 관내이식에 의해 생산된 칡소의 호반모 비율 58.1%(68/117두)보다 다소 높은 경향이었다. 2. 성별에 따른