Comparison of long noncoding RNA between muscles and adipose tissues in <i>Hanwoo</i> beef cattle

Choi, Jae-Young,Shin, Donghyun,Lee, Hyun-Jeong,Oh, Jae-Don ZOOLOGICAL SOCIETY OF KOREA 2019 ANIMAL CELLS AND SYSTEMS Vol.23 No.1

<P><B>ABSTRACT</B></P><P>Long noncoding RNAs (lncRNAs) regulate the expression of mRNA and can affect various biological processes and phenotypes. Currently, studies of lncRNAs in cattle are under way, but their exact function for several tissues has not yet been established. <I>Hanwoo</I> cattle (<I>Bos taurus coreanae</I>) have inhabited the Korean peninsula for about 6000 years and are one of the representative domesticated animals in Korea. As a result of intensive breeding, the meat of <I>Hanwoo</I> cattle is high in marbling content and is preferred by Koreans and other East Asian people. In this study, the expression of lncRNAs was identified in 36 samples from skeletal muscle and three adipose tissues (intramuscular, subcutaneous, and omental) of nine <I>Hanwoo</I> individuals. We identified 76 tissue-specific lncRNAs for each of the four tissues using the differences in expression levels. Through QTL information, we could identify 12 lncRNAs associated with shear force and six lncRNAs associated with body weight, which are two important traits in the <I>Hanwoo</I> population breeding strategy. By the physical position comparison of lncRNA and Bovine transcripts information, we could identify 11 lncRNAs that were in bovine transcripts, and four of the 11 genes related to transcripts of lncRNAs were biologically associated with muscle function. We believe this <I>Hanwoo</I> lncRNAs study will help reveal the lncRNA role in the physiological mechanisms of these four tissues.</P>

GM1 Induced the inflammatory response related to the Raf-1/MEK1/2/ERK1/2 pathway in co-culture of pig mesenchymal stem cells with RAW264.7

Kwak, Dong Hoon,Seo, You Na,Lee, Ju Hyoung,Park, Soon Ju,Cho, Young Ho,Kim, Ji-Su,Kim, Sun-Uk,Choo, Young-Kug ZOOLOGICAL SOCIETY OF KOREA 2018 ANIMAL CELLS AND SYSTEMS Vol.22 No.3

<P><B>ABSTRACT</B></P><P>Pig-human xenotransplantation can trigger cell-mediated immune responses. We explored the role of gangliosides in inflammation related to immune rejection in xenotransplantation. Co-culture of xenogeneic cells (pig-MSCs and RAW264.7) was used to emulate xenotransplantation conditions. MTT assay results indicated that cell viability was significantly decreased in pADMSCs co-cultured with RAW264.7 cells. GM1 and GM3 were highly expressed in pADMSCs co-cultured with RAW264.7 cells. pADMSCs co-cultured with RAW264.7 cells strongly expressed pro-inflammatory proteins such as COX-2, iNOS, p50, p65, pIκBα, and TNF-α. GM1-knockdown pADMSCs co-cultured with RAW 264.7 cells did not show significantly altered cell viability, but pro-inflammatory proteins were markedly inhibited. Co-culture of pADMSCs with RAW264.7 cells induced significant phosphorylation (p) of JNK1/2 and pERK1/2. However, pERK1/2 and pJNK1/2 were decreased and MEK1/2 and Raf1 were suppressed in GM1-knockdown pADMSCs co-cultured with RAW264.7 cells. Thus, the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways were significantly upregulated in response to increases of GM1 in co-cultured xenogeneic cells. However, the inflammatory response was suppressed in co-culture of GM1-knockdown pADMSCs with RAW264.7 cells via down-regulation of the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways. Therefore, the ganglioside GM1 appears to play a major role in the inflammatory response in xenotransplantation via the Raf-1/MEK1/2/ERK1/2 and JNK1/2 pathways.</P>

Effect of increased <i>p</i> CO ₂ in seawater on survival rate of different developmental stages of the harpacticoid copepod <i>Tigriopus japonicus</i>

Oh, Je Hyeok,Kim, Dongsung,Kim, Tae Won,Kang, Teawook,Yu, Ok Hwan,Lee, Wonchoel ZOOLOGICAL SOCIETY OF KOREA 2017 ANIMAL CELLS AND SYSTEMS Vol.21 No.3

<P><B>ABSTRACT</B></P><P>The rapid increase in carbon dioxide levels in seawater is causing ocean acidification and is expected to have significant effects on marine life. To explore the ability of the harpacticoid copepod <I>Tigriopus japonicus</I> to adapt to an increased concentration of dissolved carbon dioxide (CO<SUB>2</SUB>) in seawater, we compared the survival rates of adult and nauplius stages at 400, 1000, and 1550 ppm <I>p</I>CO<SUB>2</SUB> over a 14-day period. The survival rate of <I>T. japonicus</I> dramatically decreased over time with increase in <I>p</I>CO<SUB>2</SUB> concentration. At 1550 ppm, the survival rate showed a decrease of more than 20% at the end of the experimental period over that at 400 ppm. Furthermore, the survival rate decreased by a greater amount at all concentrations in nauplii than in adults, with a greater effect in wild-collected specimens than in culture-derived individuals. The results suggest that future ocean acidification may negatively influence the sustainability of <I>T. japonicus</I> and thus may eventually influence benthic ecosystems.</P>

Protective effects of ginsenoside Rg2 and astaxanthin mixture against UVB-induced DNA damage

Chung, Yu Heon,Jeong, Seul A.,Choi, Hyun Seok,Ro, Seungil,Lee, Jung Sup,Park, Jong Kun ZOOLOGICAL SOCIETY OF KOREA 2018 ANIMAL CELLS AND SYSTEMS Vol.22 No.6

<P><B>ABSTRACT</B></P><P>Ultraviolet B (UVB) radiation induces skin damage, skin matrix degradation, and wrinkle formation through photochemical reaction and oxidative stress. Therefore, protecting the skin from UVB can prevent skin aging. In this study, we investigated the effects of a mixture (RA) of Rg2, a ginsenoside, and astaxanthin, an antioxidant, on the responses of HaCaT cells exposed to UVB (700 J/m<SUP>2</SUP>). The cells were incubated for 24 h after UVB exposure and cell viability was determined by MTT assay. UVB decreased cell viability by 60% compared to that of untreated control cells, whereas RA increased cell viability in a concentration-dependent manner, and this increase was significantly higher than that in the single treatment groups. Further, UVB increased the levels of DNA lesions such as cyclobutane pyrimidine dimer (CPD) and 8-hydroxyguanine (8-OHdG). Conversely, RA decreased both CPD and 8-OHdG levels in a concentration-dependent manner. UVB exposure also increased phosphorylation of ataxia-telangiectasia mutated (ATM) protein kinase and p53 and subsequently increased the levels of GADD45α, p21, and matrix metalloproteinases (MMPs)-3, -9, and -13. Additionally, UVB exposure decreased the level of COL1A1. However, RA treatment decreased the levels of p-ATM, p-p53, GADD45α, p21, MMP-3, -9, and -13 and increased the level of COL1A1 in a concentration-dependent manner. These results suggest that RA reduces UVB-induced cytotoxicity and genotoxicity through up-regulation of DNA repair via the combined effects of Rg2 and astaxanthin.</P>

Identification of molecular markers distinguishing adult neural stem cells in the subventricular and subcallosal zones

Kim, Joo Yeon,Shaker, Mohammed R.,Lee, Ju-Hyun,Lee, Boram,Kim, Hyun,Sun, Woong ZOOLOGICAL SOCIETY OF KOREA 2017 ANIMAL CELLS AND SYSTEMS Vol.21 No.3

<P><B>ABSTRACT</B></P><P>Neural stem cells (NSCs) in the adult subventricular zone (SVZ) are regionally specified and have distinct molecular gene expression signatures. Recently, we identified the subcallosal zone (SCZ) as a novel brain region where adult NSCs maintain and spontaneously produce neuroblasts. In an attempt to isolate genes specifically expressed in the SCZ or SVZ, microarray analyses of their differentially expressing transcripts were done. The comparison between neurospheres generated from SVZ and SCZ revealed differential expression >1.5-fold in two groups in only 83 genes, representing <0.03% of the genes examined, suggesting that these two populations are largely similar. The differential expression patterns SCZ and SVZ genes were confirmed by RT-PCR and Western blots. The selective expressions of two genes (<I>CRBP1</I>, <I>HMGA1</I>) in SVZ-NSCs were further confirmed by immunohistochemistry. These molecular markers could be useful for further molecular and cellular characterization of NSCs.</P>

Fine structural reconstruction on the testicular cyst of the furrow orb weaver, Larinioides cornutus by 3D volume rendering

Kim, Hoon,Seo, Jae-Hwi,Kim, Kyo-Jin,Chung, Kyung-Hoon,Moon, Myung-Jin ZOOLOGICAL SOCIETY OF KOREA 2016 ANIMAL CELLS AND SYSTEMS Vol.20 No.5

Morphological study on spermatids and spermatozoa have long been performed regarding various changes of cell organelles during spermiogenesis as a potential phylogenetic inference. Based on the fact that the number of germ cells per cyst increases according to a geometric series, knowing the exact number of germ cells in a certain stage may lead to the total number of sperms produced per cyst. In spiders, however, the entire process takes place in a cyst represented by a spermatogonium, producing sperms in spherical shape. It is very difficult to count the exact number of germ cells produced per cyst through a 2D image analysis. Therefore, we applied a 3D image of testicular cyst of an orb-weaving spider to visualize the exact number of germ cells produced from a cyst. In this study, 2D images obtained from serially sectioned micrographs were scanned precisely and reconstructed using a 3D-rendering technique. Finally, this research reveals that the exact number of spermatozoa produced each cyst in Larinioides cornutus appeared to be 128 (<TEX>$2^7$</TEX>), which indicates that a single spermatogonium undergoes five mitotic divisions and two maturing divisions (meiosis) to produce final spermatozoa.

Optimization of episomal reprogramming for generation of human induced pluripotent stem cells from fibroblasts

Bang, Jin Seok,Choi, Na Young,Lee, Minseong,Ko, Kisung,Lee, Hye Jeong,Park, Yo Seph,Jeong, Dahee,Chung, Hyung-Min,Ko, Kinarm ZOOLOGICAL SOCIETY OF KOREA 2018 ANIMAL CELLS AND SYSTEMS Vol.22 No.2

<P><B>ABSTRACT</B></P><P>Generation of induced pluripotent stem cells (iPSCs) by defined factors (<I>OCT4</I>, <I>SOX2</I>, <I>C-MYC</I>, and <I>KLF4</I>) from various human primary cells has been reported. Human fibroblasts have been widely used as a cellular source in reprogramming studies over recent decades. The original method of iPSC generation uses retro- or lentivirus vectors that require integration of viral DNA into the target cells. The integration of exogenous genes encoding transcription factors (<I>OCT4</I>, <I>SOX2</I>, <I>C-MYC</I>, and <I>KLF4</I>) can be detected in iPSCs, raising concern about the risk of mutagenesis and tumor formation. Therefore, stem cell therapy would ideally require generation of integration-free iPSCs using non-integration gene delivery system such as Sendai virus, recombinant proteins, synthetic mRNA, and episomal vectors. Several groups have reported that episomal vectors are capable of reprogramming human fibroblasts into iPSCs. Although vector concentration and cell density are important in the episomal vector reprogramming method, optimization of this method for human fibroblasts has not been reported. In this study, we determined optimal conditions for generating integration-free iPSCs from human fibroblasts through the use of different concentrations of episomal vectors (<I>OCT4</I>/<I>p53</I>, <I>SOX2</I>/<I>KLF4</I>, <I>L-MYC</I>/<I>LIN28A</I>) and different plating cell density. We found that optimized vector concentration and cell density accelerate reprogramming and improve iPSC generation. Our study provides a detailed stepwise protocol for improved generation of integration-free iPSCs from human fibroblasts by transfection with episomal vectors.</P>

Potential use of transgenic domestic pigs expressing recombinant human erythropoietin in diabetes translation research

Baek, Sun-Young,Chung, Hak-Jae,Kim, Kyung-Woon,Cho, Kyu-Ho,Choi, Inchul,Lee, Hoon-Taek ZOOLOGICAL SOCIETY OF KOREA 2019 ANIMAL CELLS AND SYSTEMS Vol.23 No.1

<P><B>ABSTRACT</B></P><P>Recently, diabetes mellitus (DM) has shown rapid global increases with about five million deaths annually. Animal models are imperative to understand disease mechanisms and develop diagnostic, preventive, and therapeutic interventions in translational research. Rodent and mini-pig models have been established and widely used for DM research. However, domestic pig models are limited in spite of advantages such as pharmacokinetic and physiopathological availability. This study examines the potential use of domestic pigs expressing recombinant human erythropoietin (rhEPO) as disease and therapeutic response models for DM. We previously generated transgenic pigs (<I>n</I> = 16, EPO Tg) in which rhEPO was expressed and circulated in all organs. Thirty-two pigs, including 16 controls, were fed high fat (HF) diets for 42 weeks. Subsequently, blood samples for chemical and metabolic analysis were collected after fasting for 24 h and glucose loading for oral glucose tolerance tests (OGTTs). We found increased activation of the PI3 K/Akt signaling pathway under hypoxic conditions after rhEPO treatment, and HF diet-inducible-obesity in the EPO Tg and control pigs. OGTTs showed lower fasting glucose levels in the EPO Tg pigs than in controls before and after the HF diet, suggesting that rhEPO may affect glucose concentrations. Insulin and C-peptide concentrations responded slowly to glucose administration and returned to initial levels after 2 h. The blood test results suggest that EPO might affect metabolic and chemical components such as glucose, high-density lipoprotein, glucagon, triglyceride, and free fatty acid. Our findings support the use of rhEPO transgenic domestic pigs as model animals for translational DM research.</P>

Molecular Evolution of ACTIN RELATED PROTEIN6, a Component of SWR1 Complex in Arabidopsis

Park, C.,Nam, M.S.,Choi, K.,Lee, I. ZOOLOGICAL SOCIETY OF KOREA 2008 Animal cells and systems Vol.12 No.suppl

Filamin B Serves as a Molecular Scaffold for Type I Interferon-induced JNK Signaling Pathway

Lee, J.Y.,Jeon, Y.J.,Yu, K.R.,Chung, C.H. ZOOLOGICAL SOCIETY OF KOREA 2008 Animal cells and systems Vol.12 No.suppl