Characterization of Quantitative Trait Loci (QTL) for Growth using Genome Scanning in Korean Native Pig

Lee, H.K.,Choi, I.S.,Choi, B.H.,Kim, T.H.,Jung, I.J. The Korean Society of Animal Reproduction 2004 Reproductive & developmental biology Vol.28 No.2

Molecular genetic markers were genotyped used to detect chromosomal regions which contain economically important traits such as growth traits in pigs. Three generation resource population was constructed from a cross between the Korean native boars and Landrace sows. A total of 193 F2 animals from intercross of F1 were produced. Phenotypic data on 7 traits, birth weight, body weight at 3, 5, 12, 30 weeks of age, live empty weight were collected for F2 animals. Animals including grandparents (F0), parents (F1), offspring (F2) were genotyped for 194 microsatellite markers covering from chromosome 1 to 18. Quantitative trait locus analyses were performed using interval mapping by regression under line-cross model. To characterize presence of imprinting, genetic full model in which dominance, additive and imprinting effect were included was fitted in this analysis. Significance thresholds were determined by permutation test. Using imprinting full model, four QTL with expression of imprinted effect were detected at 5% chromosome-wide significance level for growth traits on chromosome 1, 5, 7, 13, 14, and 16.

Application of Intracytoplasmic Sperm Injection in Mammals

Li, Xiao Xia,Lee, Kyung Bon,Lee, Ji Hye,Kim, Keun Jung,Park, Kang Sun,Kim, Min Kyu The Korean Society of Animal Reproduction 2013 Reproductive & developmental biology Vol.37 No.2

For more than two decades, the intracytoplasmic sperm injection (ICSI) technique has been used as a valuable tool to provide opportunities for studying fertilization, treating human infertility, and producing transgenic animals. Not only in facilitating fertilization but also in propagating mammalian species, ICSI has enhanced the potential of assisted reproductive technologies in human. Polyspermic fertilization has been one of major problems in pig reproduction, but the ICSI helped to solve the problem, and used widely to generate transgenic piglets. Although the ICSI technique is considered to be a very useful tool in assisted reproductive technologies, including generation of transgenic animals, there are some disadvantages using the technique. In this review, we describe the ICSI technique and its application in animal production and human infertility, and discuss advantage and disadvantage of the technique in mammals.

A Role of Unsaturated Fatty Acid in Animal Reproductive Cells and Biology

Hwangbo, Yong,Kim, Hwa-Young,Lee, Yu-Rim,Lee, Seung Tae,Lee, EunSong,Cheong, Hee-Tae,Yang, Boo-Keun,Park, Choon-Keun The Korean Society of Animal Reproduction 2016 Reproductive & developmental biology Vol.40 No.2

As a one of unsaturated fatty acid, polyunsaturated fatty acids (PUFAs) have multiple actions: as precursor of prostaglandins (PGs), steroid hormone synthesis and energy production in animal reproduction. PUFAs, which include omega-3 (n-3) and omega-6 (n-6), are derived from the diet and changed by diet, species, breed and season. The plasma membrane of spermatozoa in mammals contain various PUFAs. These composition of PUFAs regulate the membrane fluidity and cause lipid peroxidation via generation of reactive oxygen species (ROS). Induced lipid peroxidation by ROS decreased viability and motility of spermatozoa, and it is reduced by addition of antioxidant and low concentration of PUFAs. Because oocytes of animal have a high lipid components, process of oocyte maturation and embryo development are influenced by PUFAs. In in vitro study, oocyte maturation, embryo development, intracellular cAMP and MAPK activity were increased by treatment of n-3 ${\alpha}$-linolenic acid (ALA) during maturation, whereas n-6 linoleic acid (LA) negatively influenced. Also, inhibition of fatty acid metabolism in oocyte influenced blastocyst formation of cattle. PGs are synthesized from PUFAs and various PUFAs influence PGs via regulation of PG-endoperoxide synthase (PTGS). Steroid hormone synthesis from cholesterol is regulated by expression of steroid acute regulator (StAR) protein and mRNA. Exogenous n-3 and n-6 PUFAs altered sex hormone in animal through stimulate or inhibit StAR activity. Because PUFAs altered PG and steroid hormone synthesis, follicular development was influenced by PUFAs. This effect of unsaturated fatty acid could provide information for improvement of reproductive ability in animals.

Gene Co-Expression Network Analysis of Reproductive Traits in Bovine Genome

Lim, Dajeong,Cho, Yong-Min,Lee, Seung-Hwan,Chai, Han-Ha,Kim, Tae-Hun The Korean Society of Animal Reproduction 2013 Reproductive & developmental biology Vol.37 No.4

Many countries have implemented genetic evaluation for fertility traits in recent years. In particular, reproductive trait is a complex trait and need to require a system-level approach for identifying candidate genes related to the trait. To find the candidate gene associated with reproductive trait, we applied a weighted gene co-expression network analysis from expression value of bovine genes. We identified three co-expressed modules associated with reproductive trait from bovine microarray data. Hub genes (ZP4, FHL2 and EGR4) were determined in each module; they were topologically centered with statistically significant value in the gene co-expression network. We were able to find the highly co-expressed gene pairs with a correlation coefficient. Finally, the crucial functions of co-expressed modules were reported from functional enrichment analysis. We suggest that the network-based approach in livestock may an important method for analyzing the complex effects of candidate genes associated with economic traits like reproduction.

Minipigs as Laboratory Animals: Facility Management and Husbandry

Koo, Ok-Jae,Jang, Goo,Lee, Byeong-Chun The Korean Society of Animal Reproduction 2012 Reproductive & developmental biology Vol.36 No.1

Minipigs are regarded as one of the most important laboratory animal in that anatomical and physiological properties are similar to human and their reproduction efficiency is relatively higher compared to other large animal species. Particularly, several diseases that cannot be mimicked in rodent models are successfully occurred or induced in pig models therefore it has been interested in a valuable model for human diseases. Pigs are also 'standard' species in xenotransplantation research. To maximize experimental outcome using minipigs, establishment and management of proper animal facility, right animal husbandry and control of pathogens are very important. In this review, we summarized several international guidelines related with minipigs published by several companies or governments and discuss optimal conditions for providing informative ideas to the researchers who want to use minipigs in their future studies.

Paternity Diagnosis using The Multiplex PCR with Microsatellite Markers in Dogs

Kim, Seung-Chang,Jang, Hong-Chul,Kim, Lee-Kyung,Lim, Da-Jeong,Lee, Seung-Hwan,Cho, Yong-Min,Kim, Tae-Hun,Seong, Hwan-Hoo,Oh, Sung-Jong,Choi, Bong-Hwan The Korean Society of Animal Reproduction 2011 Reproductive & developmental biology Vol.35 No.4

The number of abandoned dogs is increasing with the worsening of the economy and the rising of feed value. It was becoming a serious social problem because of the disease transmission and destruction of natural ecosystems by abandoned dogs been wild animal. In order to solve these problems, companion dogs necessary to secure its own genetic information and to establish the systematic tracking system. Using multiplex-PCR method with 27 microsatellite marker (MS marker) divided 3 set, various alleles occurring to 6 dog breed (Labrador Retriever, German Shepherd, English Springer Spaniel, Belgian Malinois, Jindo Dog, PoongSan Dog) make use of markers to determine allele frequency and heterozygosity. MS marker FH2834 and FH2790 have only two allele and most were found in 13 alleles at FH3381 and FH3399. Average heterozygosity of MS marker is 0.534 and especially, heterozygosity represented the highest value of 0.765 at FH3381. So, it was recognized appropriate allele frequency for individual identification and paternity diagnosis in companion dogs. Using multiplex-PCR method with MS marker, various alleles occurring to dog breed make use of markers to deter mine individual identification and paternity diagnosis, traits associated biomarkers and breed-specific marker for faster, more accurate and ways to reduce the analysis cost. Based on this result, a scientific basis was established to the existing pedigree data by applying genetics additionally. Animal registration system is expected to be conducted nationwide in future. The method expects to very useful this system.

Inhibition of Histone Deacetylase Activity Diminishes Pressure Overloaded Cardiac Hypertrophy in Mice

Hong, Yun-Kyung,Song, Jong-Wook,Lee, Sang-Kil,Lee, Young-Jeon,Rho, Gyu-Jin,Kim, Joo-Heon,Hong, Yong-Geun The Korean Society of Animal Reproduction 2011 Reproductive & developmental biology Vol.35 No.2

To explore the role of histone deactylase (HDAC) activation in an in vivo model of hypertrophy, we studied the effects of Trichostatin A (TSA). TSA subjected to thoracic aortic banding (TAB)-induced pressure stress in mice. In histological observations, TAB in treated mice showed a significant hypertrophic response, whereas the sham operation remained nearly normal structure with partially blunted hypertrophy. TSA treatment had no effect (measured as HW/BW) on sham-operated animals. TAB animals treated with vehicle manifested a robust ~50% hypertrophic response (p<0.05 vs sham). TAB mice treated with 2 mg/kg/day TSA manifested a blunted growth responses, which was significantly diminished (p<0.05) compared with vehicle-treated TAB mice. TAB mice treated with a lower dose of TSA (0.5 mg/kg/day) manifested a similar blunting of hypertrophic growth (~25% increase in heart mass). Furthermore, to determine activity duration of TSA in vitro, 1 nM TSA was added to H9c2 cells. Histone acetylation was initiated at 4 hr after treatment, and it was peak up to 18 hr, then followed by significantly reduced to 30 hr. We also analyzed the expression of p53 following TSA treatment, wherein p53 expression was elevated at 4 hr, and it was maintained to 24 hr after treatment. ERK was activated at 8 hr, and maintained till 30 hr after treatment suggesting an intracellular signaling interaction between TSA and p53 expression Taken together, it is suggested that HDAC activation is required for pressure-overload growth of the heart. Eventually, these data suggest that histone acetylation may be a novel target for therapeutic intervention in pressure-overloaded cardiac hypertrophy.

Unanticipated Gene Deletion in the Transgenic Chicken Employing Ovalbumin Promoter for Oviduct Specific Expression

Jang, Tae Young,Koo, Bon Chul,Kwon, Mo Sun,Roh, Ji Yeol,Kim, Teoan,Park, Young Sik The Korean Society of Animal Reproduction 2013 Reproductive & developmental biology Vol.37 No.3

Transgenic chickens have been spotlighted as an highly potent bioreactor for their fecundity, short generation time, and eggs associated with mass production of protein. In this study, we generated transgenic chickens exhibiting oviduct specific expression of human growth hormone fused to human transferrin for oral administration. Gene of the modified growth hormone located at downstream ovalbumin promoter (~3.6 kb) was introduced to stage X blastodermal cell employing retrovirus vector system. Several transgenic chickens were successfully generated. However, genomic analyses showed unexpected deletion within the transgene. The modification of the transgene seemed to occur during germ cell formation because the deletion was detected only from the sperm DNA of the G0 founder animal. There was no evidence of deletion in the somatic cell DNA samples of the same chicken. Consequently, same pattern of the deletion was confirmed in both somatic and germ cells of the G1 progeny.

Optimal Condition for Sperm-mediated Gene Transfer by Liposome in Pigs

Kim, Tae-Shin,Yang, Cao,Lee, Young-Seung,Park, Soo-Bong,Park, Chun-Keun,Lee, Dong-Seok The Korean Society of Animal Reproduction 2008 Reproductive & developmental biology Vol.32 No.2

Production of transgenic animals for studying specific gene has been limited due to a low efficiency, lack of skilled researchers and the need for expensive equipment. Currently, the boar spermatozoa as a vector to deliver exogenous DNA into the oocyte were used to improve the efficiency of transfection rate. In this study, we revealed that the optimal conditions for DNA uptake in spermatozoa by liposome were to 90 min of incubation, $17^{\circ}C$, $10^5$ spermatozoa, 4 ng/ml of exogenous DNA and 0.5% (v/v) liposome, without damage to fertility. In addition, the developmental rate to the blastocyst stage of embryo in control group was significantly higher than those embryos with exogenous DNA and liposome, whereas there were no significant differences in embryo development between the liposome and type of DNA. The transfection rates of embryo using treated spermatozoa with both liposome and circular DNA were higher than those using linear DNA. These findings raise the possibility thattreated spermatozoa with liposome/DNA complexes could be used in in vitro fertilization, and the exogenous DNA transferred into the oocytes. Taken together, we demonstrated that liposome a vector for the uptake of exogenous DNA in boar spermatozoa could improve the efficiency of sperm-mediated gene transfer in creating transgenic pig and the other domestic transgenic animals.

Polyclonal Antibody to a 37-kDa Recombinant Protein Derived from Bovine $20{\alpha}$-Hydroxysteroid Dehydrogenase

Naidansuren, Purevjargal,Min, Kwan-Sik The Korean Society of Animal Reproduction 2012 Reproductive & developmental biology Vol.36 No.2

We prepared the polyclonal antibody anti-$20{\alpha}$-hydroxysteroid dehydrogenase (anti-$20{\alpha}$-HSD) against the recombinant full-length protein bovine $20{\alpha}$-HSD in Escherichia coli. The specificity of anti-$20{\alpha}$-HSD was demonstrated using Chinese hamster ovary (CHO) cells transfected with recombinant bovine $20{\alpha}$-HSD and bovine placental tissues. According to western blot analysis, anti-$20{\alpha}$-HSD specifically recognizes the 37-kDa protein bovine $20{\alpha}$-HSD. The protein is not present in untransfected CHO cells. Anti-$20{\alpha}$-HSD also recognizes a specific protein in the ovaries and placenta of other animals. Immunostaining was used to detect expression of bovine $20{\alpha}$-HSD protein in the cultured luteal cells during the estrous cycle later.