Xenotransplantation is proposed as a solution to the problem of organ shortage. However, transplantation of xenogeneic organs induces an antigen-antibody reaction in α-1,3-gal structure that are not present in humans and primates, and thus complement is also activated and organs die within minutes or hours. In this study, we used FasL gene, which is involved in the immune response of NK cell, and US11, which suppresses MHC Class I cell membrane surface expression, to inhibit cell mediated rejection in the interspecific immunity rejection, and also hDAF(CD55) was introduced to confirm the response to C3 complement. These genes were tranfeced into Korean native pig fetal fibroblasts using pCAGGS vector. And cytotoxicity of NK cell and human complement was confirmed in each cell line. The US11 inhibited the cytotoxicity of NK cell and, in addition, the simultaneous expression of US11 and Fas ligand showed excellent suppress to T-lymphocyte cytotoxicity, hDAF showed weak resistance to cytotoxicity of natural killer cell but not in CD8+ CTLs. Cytotoxicity study with human complement showed that hDAF was effective for reducing complement reaction. In this studies have demonstrated that each gene is effective in reducing immune rejection.

Embryo development is very important in reproductive physiology of domestic animal experiments. Therefore, in the above experiment, we want to provide a lot of important information with regard to fertilization breeding by looking at the expression of transcription factor by early embryo development. It is known that mice affect early embryonic development of many transcription factors, many experiments are underway. Different types of mammals showed different expression patterns, thus, we used pigs, which are known to be the most similar to humans, to observe the expression of transcription factors in early embryonic development. Transcription factors were observed using CDX2, OCT4 and E-CADHERIN. CDX2 was expressed in 2 cells, OCT4 and E-CADHERIN were expressed in blastocyst. OCT4 was expressed specifically in ICM (inner cell mass) in blastocyst, and E-CADHERIN was expressed in cell wall and junction of blastocyst. These results show that CDX2, OCT4 and E-CADHERIN play an important role in early embryonic development in pigs.

Porcine litter size is a quantitative trait and its heritability is especially low. So it is necessary to identify porcine reproductive gene and protein. The establishment of pregnancy requires performance of a receptive endometrium. We analyzed the endometrial tissue protein of porcine and would find out biomarker proteins related to porcine litter size. We sorted the two groups according to litter size of porcine: a small litter size group (SLSG) (n=2) and a large litter size group (LLSG) (n=2). The porcine endometrial tissue samples were analyzed separately using 2-dimensional electrophoresis (2-DE) within the isoelectric point ranges of 3.0 to 10.0, and then differential proteins were identified using MALDI-TOF analysis. In comparison of SLSG(small litter size group) with LLSG(large litter size group), a total of 9 protein spots differentially expressed on porcine endometrial tissue 2-DE gels, among which 5 spots were up-regulated proteins as retinol dehydrogenase 16-like isoform 1, Acrosin-binding protein, alpha-N-acetylgalactosaminidase. phosphoglycerate kinase 2, Acrosin-binding protein in LLSG. And 4 spots were up-regulated proteins as phosphoglycerate kinase 2, prenylcysteine oxidase in SLSG.

The red-spotted apollo butterfly, Parnassius bremeri Bremer, 1864 (Lepidoptera: Papilionidae), has been listed as an endangered species in Korea. We developed microsatellite markers by nest-generation sequencing (NGS), selected 12 markers, and applied the markers to available South Korean populations to understand population genetic characteristics. The genotyping of 40 P. bremeri individuals from three localities in South Korea showed that at each locus, the observed number of alleles ranged from 17 to 43, the observed and expected heterozygosities were 0.84722-0.90556 and 0.76045-0.79208, respectively, and FIS was –0.155 to –0.121. STRUCTURE analysis supported the presence of two genetic pools in all three populations, although an immediate reason for this subdivision is not known. The population based FST, RST, and front wing length collectively suggest that at least Samchuk population in Gangwon Province has a significant distance. Further scrutinized analysis is undergoing.

Despite of a substantial effort, the phylogeny of Lepidoptera still has unstable taxa and unresolved relationships particularly form Apoditrysia to Ditrysia (e.g. phylogenetic position and familial relationships of Gelechioidea, the monophylies of Tineoidea and Zygaenoidea and so on). We performed phylogenetic analysis using ~13,000-bp length of mitogenome sequences form 50 species (with seven species in two superfamilies form this study) representing 13 superfamilies to improve the lepidopteran phylogeny from Apoditrysia to Ditrysia. A close relationship of Gelechioidea to Obtectomera than to apoditrysian taxa was noteworthy, although the nodal support is very weak (ML, 23%; BI, BPP = 0.94). Monophyly of Zygaenoidea without Epipyropidae (Epipomponia nawai) was also noteworthy in spite of weak nodal support (ML, 64%; BI, 1.0). Scrutinized analysis is currently underway.

We sequenced the complete mitochondrial genome (mitogenome) of Vespa velutina nigrithorax and V. ducalis (Hymenoptera: Vespidae). The genomes were 16,475-bp and 15,779-bp long and contained typical sets of genes. The V. velutina and V. ducalis A+T-rich region was 132-bp long and 166-bp long and was the shortest of all sequenced Vespoidea genomes. Start and stop codons in several Vespa species—including V. velutina and V. ducalis—were diversified, despite these species belonging to the same genus. In comparison with the ancestral mitogenomes, Vespa mitogenomes showed substantial gene rearrangement; however, we detected no gene rearrangement among Vespa species. We conducted phylogenetic reconstruction based on concatenated sequences of 13 PCGs and two rRNAs in available species of Vespoidea—22 species in six subfamilies in two families (Vespidae and Formicidae). The Bayesian inference and maximum likelihood (ML) methods revealed that each family formed strong monophyletic groups.

α-solanine is toxic to human health by disturbing digestive and central nervous systems. However, little information has been focused on investigated with respect to α-solanine influence in mammal oocyte maturation and quality. In this study, we investigated the effects of α-solanine on oocyte maturation, quality and possible molecular mechanisms in a pig model. Porcine Cumulus-oocyte complexes (COCs) were treated with increasing concentration (0, 1, 10, 20, 50 μM) of α-solanine subjected to further in vitro maturation culture. The result showed that α-solanine significantly inhibited cumulus cells expansion and increased oocyte death rates when the concentration of α-solanine more than 10 μM. After cell cycle and cytoskeleton analysis, the results showed that α-solanine (10 μM) disturbed meiotic resumption, increased abnormal spindle formation and cortical granules (CGs) distribution rates when compared with the untreated group. α-solanine (10 μM) triggered autophagy by increasing the expression of autophagy-related genes (LC3, ATG7, LAMP2) and accumulation of LC3-specific puncta (an autophagy maker). TUNEL staining assay showed that α-solanine significantly increased apoptosis in porcine oocytes confirmed by up-regulated the levels of BAX and CAPS3 genes. Further study revealed that exposure α-solanine (10 μM) to porcine oocytes induced ROS generation, reduced mitochondrial membrane potential. In addition, our results suggested that α-solanine (10 μM) significantly increased the levels of H3K36me3 and H3K27me3 in porcine oocytes. Taken together, these data indicated that α-solanine toxic impaired oocyte maturation and quality by inhibited cumulus cells expansion, increased abnormal spindle and CGs distribution rates, triggered autophagy/apoptosis occur, accumulated ROS, decreased mitochondrial membrane potential, and changed epigenetic modifications.

In pig, more than half of the recovered cumulus cell-oocyte complexes (COCs) have one or two layers of cumulus cells and are considered morphologically poor. If we could take full advantage of these poor quality COCs, we could potentially improve the efficiency of in vitro embryo production. During in vitro maturation, although some maturation factors are transmitted bidirectionally between the oocyte and cumulus cells of the same COC, transmission also occurs between different COCs. We hypothesized that morphologically poor COCs fail to undergo complete oocyte maturation due to their insufficient secretion of maturation factors. Here, we investigated whether co-culture with morphologically good COCs (having three or more layers of cumulus cells) could improve the maturation and utilization rates of morphologically poor COCs. Our results revealed that the oocyte maturation rate, glutathione level, embryo development capacity, blastocyst quality, and cumulus cell gene expression levels of BCL-2 and PCNA were similar in the co-culture and good quality-groups, and that these levels were all significantly higher than those in the poor quality-group. Our results strongly suggest that the co-culture strategy greatly improved the utilization rate of morphologically poor COCs without reducing their capacity for maturation and subsequent development.

Early pregnancy results in th production of various signal molecules such as steroids, prostaglandins, and many protein factors. The proteins especially produced by the placenta have been used to detect pregnancy for many years in other species. More recently, pregnancy-specific protein B, which is a placental glycoprotein can be measured by RIA or proteomic methods in serum of pregnant cow. And 2D Fluorescence difference gel electrophoresis (DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. For this reason, we are analyzed serum of bovine. The purpose of this study was to apply DIGE technique for identification of bovine pregnancy-specific proteins using bovine pregnant and non-pregnant serum samples. Serums of 2 pregnant Holstein dairy cattle at day 21 after AI and those of 2 non-pregnant were used in this study. The molecular weight and charge matched cyanine dyes enable pre-electrophoretic labeling of non-pregnancy and pregnant serum proteins which are then mixed and labeled with Cy2 were used as an internal standard. Two pools of proteins are labeled with Cy3 and Cy5 fluorescent dyes, respectively. Labeled proteins with Cy2, Cy3 and Cy5 mixed together and separated in same gel and then were detected by fluorescence image analyzer. The 2D DIGE analysis using fluorescence CyDye flour showed higher sensitivity and better reproducible results than conventional 2D gel electrophoresis. Approximately 1,500 protein spots were detected by 2D DIGE. The differentially expressed proteins were identified by MALDI-TOF Mass spectrometer. Total 16 protein spots differentially expressed in the pregnant serum were detected, among which 7 spots were up-regulated proteins identified as conglutinin precursor, modified bovine fibrinogen, IgG1 etc, and 6 spots were down-regulated proteins identified as hemoglobin, complement component 3, bovine fibrinogen, IgG2a etc. These results indicated that DIGE system could be advantageous for the analysis of serum proteomics diversified by physiological conditions.