• Title/Summary/Keyword: Roles in reproduction

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Zebrafish as a Tool for Function Genomics (제브라피쉬를 이용한 새로운 유전자의 발굴 및 기능분석)

  • Kim Hyun Taek;Kim Cheol Hee
    • Development and Reproduction
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    • v.7 no.2
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    • pp.69-80
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    • 2003
  • The zebrafish(Danio rerio) is a pre-eminent vertebrate model system for clarification of the roles of specific genes and signaling pathways in development. We show examples of positional cloning in two developmental mutants in zebrafish. headless: The severe head defects in headless(hdl) mutants are due to a mutation in T-cell factor-3(Tcf3). Loss of Tcf3 function in the hdl mutant reveals that Hdl represses Wnt target genes. The results provide genetic evidence that a component of the Wnt signaling pathway is essential in vertebrate head formation and patterning. mind bomb: Reduced lateral inhibition in mind bomb(mib) mutants permits too many neural precursors to differentiate as neurons. Positional cloning of mib revealed that it is a gene in the Notch pathway that encodes a ubiquitin E3 ligase. Mib interacts with the intracellular domain of Delta to promote its internalization. The results suggest a model for Notch activation where the Delta-Notch interaction is followed by endocytosis of Delta and transendocytosis of the Notch extracellular domain by the signaling cell.

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Four Members of Heat Shock Protein 70 Family in Korean Rose Bitterling (Rhodeus uyekii)

  • Kim, Jung Hyun;Dong, Chun Mae;Kim, Julan;An, Cheul Min;Baek, Hae Ja;Kong, Hee Jeong
    • Development and Reproduction
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    • v.19 no.3
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    • pp.135-144
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    • 2015
  • Heat shock protein (HSP) 70, the highly conserved stress protein families, plays important roles in protecting cells against heat and other stresses in most animal species. In the present study, we identified and characterized four Hsp70 (RuHSP4, RuHSC70, RuHSP12A, RuGRP78) family proteins based on the expressed sequence tag (EST) analysis of the Korean rose bitterling R. uyekii cDNA library. The deduced RuHSP70 family has high amino acid identities of 72-99% with those of other species. Phylogenetic analysis revealed that RuHsp70 family clustered with fish groups (HSP4, HSC70, HSP12A, GRP78) proteins. Quantitative RT-PCR analysis showed the specific expression patterns of RuHsp70 family members in the early developmental stages and several tissues in Korean rose bitterling. The expression of 4 groups of Hsp70 family was detected in all tested tissue. Particularly, Hsp70 family of Korean rose bitterling is highly expressed in hepatopancreas and sexual gonad (testis and ovary). The expression of Hsp70 family was differentially regulated in accordance with early development stage of Rhodeus uyekii.

Expressional Modulation of Connexin Isoforms in the Initial Segment of Male Rat treated with Estradiol Benzoate or Flutamide

  • Lee, Ki-Ho
    • Development and Reproduction
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    • v.18 no.4
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    • pp.293-300
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    • 2014
  • Direct cell-cell communication through connexin (Cx) complexes is a way to achieve functional accordance of cells within a tissue or an organ. The initial segment (IS), a part of the epididymis, plays important roles in sperm maturation. Steroid hormones influence on expression of a number of genes in the IS of adult animals. However, developmental effect of sex hormones on the gene expression in the IS has not been examined. In this study, estradiol benzoate (EB, an estrogen agonist) or flutamide (Flu, an androgen antagonist) was exogenously administrated at 1 week of postnatal age, and expressional changes of Cx genes in the IS were determined at 4 months of age by a quantitative real-time PCR analysis. Treatment of EB at $0.015{\mu}g/kg$ body weight (BW) increased expression of Cx30.3, 31.1, and 43 genes. However, treatment of 1.5 mg EB/kg BW resulted in expressional decreases of Cx31, 32, and 45 genes and caused increases of Cx30.3 and 43 gene expression. Significant decreases of Cx31, 31.1, 32, 37, and 45 gene expression were detected with a treatment of $500{\mu}g\;Flu/kg$ BW, while expression of Cx43 gene was significantly increased with a treatment of $500{\mu}g\;Flu/kg$ BW. A treatment of $50{\mu}g\;Flu/kg$ BW led to significant increases of Cx30.3, 32, 37, 40, and 43 gene expression. These findings imply that exogenous exposure of steroidal hormones during the early developmental period would result in aberrant expression of Cx genes in the adult IS.

Identification and Characterization of a Putative Basic Helix-Loop-Helix (bHLH) Transcription Factor Interacting with Calcineurin in C. elegans

  • Lee, Soo-Ung;Song, Hyun-Ok;Lee, Wonhae;Singaravelu, Gunasekaran;Yu, Jae-Ran;Park, Woo-Yoon
    • Molecules and Cells
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    • v.28 no.5
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    • pp.455-461
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    • 2009
  • Calcineurin is a $Ca^{2+}$/Calmodulin activated Ser/Thr phosphatase that is well conserved from yeast to human. It is composed of catalytic subunit A (CnA) and regulatory subunit B (CnB). C. elegans homolog of CnA and CnB has been annotated to tax-6 and cnb-1, respectively and in vivo function of both genes has been intensively studied. In C. elegans, calcineurin play roles in various signaling pathways such as fertility, movement, body size regulation and serotonin-mediated egg laying. In order to understand additional signaling pathway(s) in which calcineurin functions, we screened for binding proteins of TAX-6 and found a novel binding protein, HLH-11. The HLH-11, a member of basic helix-loop-helix (bHLH) proteins, is a putative counterpart of human AP4 transcription factor. Previously bHLH transcription factors have been implicated to regulate many developmental processes such as cell proliferation and differentiation, sex determination and myogenesis. However, the in vivo function of hlh-11 is largely unknown. Here, we show that hlh-11 is expressed in pharynx, intestine, nerve cords, anal depressor and vuvla muscles where calcineurin is also expressed. Mutant analyses reveal that hlh-11 may have role(s) in regulating body size and reproduction. More interestingly, genetic epistasis suggests that hlh-11 may function to regulate serotoninmediated egg laying at the downstream of tax-6.

Development and Characterization of Hyperglycosylated Recombinant Human Erythropoietin (HGEPO)

  • JarGal, Naidansuren;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • v.33 no.2
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    • pp.77-83
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    • 2009
  • Erythropoietin (EPO), a glycoprotein hormone produced from primarily cells of the peritubular capillary endothelium of the kidney, is responsible for the regulation of red blood cell production. We have been investigating the roles of glycosylation site added in the biosynthesis and function of recombinant protein. We constructed three EPO mutants ($\Delta$69, $\Delta$105 and $\Delta$69,105), containing an additional oligosaccharide chains. EPOWT and EPO$\Delta$69 were effectively expressed in transient and stably transfected CHO-K1 cell lines. But, it wasn't detected any protein in the culture medium of EPO$\Delta$105 and EPO$\Delta$69,105 mutants. The growth and differentiation of EPO-dependent human leukemic cell line (F36E) were used to measure the cytokine dependency and in vitro bioactivity of rec-hEPO. MTT assay values were increased by survival of F36E cells at 24h. To analysis biological activity in vivo, two groups of ICR-mice (7 weeks old) were injected subcutaneously with 10 IU per mice of rec-hEPO molecules on days 0 and 2. Red blood cell and hematocrit values were measured on 6 days after the first injection. The hematocrit values were remarkably increased in all treatment groups. The pharmacokinetic analysis was also affected in the mice injected with rec-hEPO molecules 2.5 IU by tail intravenous. Protein samples were detected by Western blotting. An EPO$\Delta$69 protein migrated as a broad band with an average apparent molecular and detected slightly high band. Enzymatic N-deglycosylation resulted in narrow band and was the same molecular size. The biological activity of EPO$\Delta$69 was enhanced to compare with wt-hEPO. The half-life was longer than wt-hEPO. The results suggest that hyperglycosyalted recombinant human erythropoietin (EPO$\Delta$69) may have important biological and therapeutic good points.

Functioning of the Geoecosystem for the West Side of Admiralty Bay (King George Island, Antarctica): Outline of Research at Arctowski Station

  • Rakusa-Suszczewski, Stanislaw
    • Ocean and Polar Research
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    • v.25 no.4
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    • pp.653-662
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    • 2003
  • Changes in the area of geo-ecosystem $(62^{\circ}09'S,\;58^{\circ}31'w)$ reflect climatic changes in the South Shetland Islands. Air temperature and deglaciation will increase. The ice-free space area at the SSSS 8- (ASPA 121) site has enlarged threefold during the last 21 years, thus creating conditions for inhabitation and succession. Wind, water and snow play important roles in transportation of geochemical components. They distribute nutrients, mineral substances, seeds, fragments of plants and animals, etc. Plant and animal colonization is patchy and it happens at random in an 'island' - like manner. The colonization pattern is dependant, to a high degree on physical factors. The newly uncovered ice-free areas are at first inhabited by a vascular plant known as the Deschampsia antarctica. The border of the land-oasis with Admiralty Bay is the place where the processes related to animal feeding at the sea and reproduction on the land take place. Bird colonies and pinniped lairs form centers of fertilization surrounded by high chemical gradients dependent on the direction of the flow of nutrients $(e.g.\;NH_4)$. During the last 25 years, the numbers of penguins in this region have decreased, and thus the amount of materials excreted on land has diminished. The numbers of fur seals change in multi-annual cycles, and their migration into this area is related to the E1 $Ni\~{n}o$ phenomenon. The numbers of elephant seals in the area did not change. Organic matter deposited by the sea onto the shore are a source of nutrients and deficient chemical elements on land. Mineral matter is washed out into the waters of Admiralty Bay. These processes change seasonally, and multi annually. Negative effects on the environment at Arctowski Station induced by man are slight, but noticeable nevertheless. Physical processes have the largest influence on the living conditions and distribution of plants and animals, and as a consequence, on the functioning of the geo-ecosystem in the coastal-shore zone of the Maritime Antarctic.

Prostaglandins Involving in Blastocyst Development through Calcium Mediated Signaling Pathway

  • Lee, Ja-Myong;Kwon, Hyuck-Chan;Lee, Seung-Jea;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.15 no.1
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    • pp.17-24
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    • 2011
  • Lipid metabolites involved in cellular regulation as signaling mediators. Prostaglandins (PGs), metabolites of lipid are involved to pregnancy at the time of implantation but the functional roles of PGs on embryo development are still controversy and largely unknown. In previous report, the levels of $PGE_2$ and $PGF_{2a}$ at embryos of morula stage and blastocyst stage were explored (Cheon et al., 1998). In this study, the previous suggestion was confirmed and the possible downstream mediator of prostaglandin $E_2$ and prostaglandin $F_{2a}$ on the expansion and hatching of mouse embryo was examined. As expected, developmental rate of the blastocyst to expanded stage was a concentration-response curve that showed the highest expansion rate at 10 ${\mu}M$ $PGE_2$, but at 100 ${\mu}M$ $PGE_2$, the rate was decreased. In contrast to the $PGE_2$, $PGF_{2a}$ stimulated expansion without toxicity at highest concentration. Cotreatment of PGs with indomethacin overcame the inhibitory effects of indomethacin in expansion. Exogenous PGs also improved the development of expanded embryos to the hatching stage. Besides, PGs receptors' transcripts detected at blastocyst. $PGE_2$ was caused of calcium fluctuation in the blastocyst but $PGF_{2a}$ did not. The changes of intracellular calcium concentration were different between indomethacin pretreated embryos and non-treated embryos. Based on these results it is suggested that PGs work as paracrine and/or autocrine factors through calcium and the others which were not identified in this study.

Differential Expression Analysis of Candidate Genes Related with Growth according to Dietary Supplementation of Curcuma longa in Chickens

  • Park, Sun-Ae;Kim, Lee-Kyung;Park, Chang-Min;Kim, Seung-Chang;Lee, Seung-Hwan;Lee, Ji-Woong;Choi, Bong-Hwan
    • Reproductive and Developmental Biology
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    • v.37 no.4
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    • pp.199-203
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    • 2013
  • This experiment was conducted to investigate the genetic effects of candidate genes on the growth of spleen and liver tissues using dietary Curcuma longa (C. longa) supplementation. Expression analyses of candidate genes regarding animal growth was performed in order to determine the factors affecting the growth related to immune components of Curucumin, Turmerone, and Zingiberene as the bile secretion Paratolyl methyl carbinol (PTMC). The animals were divided into four groups of five chicks supplied with experimental diets of C. longa at 0.25, 0.5 and 1% and controls. The 19 growth-related genes were known to cell maturation, differentiation significant expression patterns in this analysis. Expression of growth response-related genes in chicks supplemented with 1% of C. longa showed better growth performance than chicks with 0.25 and 0.5% in spleen (p<0.05). The IGF1, MSTN, POU1F1, ADCYAP1 gene were known to central roles in mediating gonadotropin function, regulating steroidogenesis and promoting oocyte growth and maturation. Sex steroids, androgen and estrogen can affect sex differentiation and also can affect muscle development. On the other hand, GHSR and FABP3 gene showed significant expression patterns in this analysis. The results would be used as basic information for the variation of growth-related genes expression on the cell growth, sex cell growth, and sex hormones according to dietary supplementation with C. longa in chickens.

Association of Endocrine Factors (Insulin-Like Growth Factor-II and Binding Protein-3) with Litter Size in Pigs

  • Yun, J.S.;Kang, W.J.;Seo, D.S.;Park, S.S.;Hong, K.C.;Lee, C.Y.;Ko, Y.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.3
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    • pp.307-315
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    • 2001
  • Litter size has been one of the important economic traits in porcine reproduction. The insulin-like growth factor (IGF) system has been shown to mediate actions of the steroid hormone or to synergize with other endocrine factors so that it consequently plays roles in reproductive processes, including ovulation, implantation, maintenance of pregnancy, and fetal development. However, the effect of the serum IGF system on porcine litter size has not been deeply studied. Therefore, this study was conducted to relate serum IFG-II concentration and IGF binding protein-3 (IGFBP-3) expression with porcine litter size. Moreover, the possible association of those with estrogen receptor (ER) as a candidate gene for litter size was investigated. Swine were separated into two groups showing high and low litter sizes, and sera were collected from sows in the estrous cycle to postnatal growth of their female progeny. Serum IFG-II concentration was measured by radioimmunoassay and IGFBP-3 expression was detected by Western ligand blotting. During the estrous cycle, IGFBP-3 expression in both groups decreased moderately from metestrus to estrus, but IFG-II concentration showed a reverse pattern. Also, IFG-II concentration and IGFBP-3 expression decreased gradually as pregnancy proceeded. Unlike IGFBP-3, IFG-II decreased moderately as newborn pigs grew. Significant differences in serum IFG-II amount between the two groups were detected at 60 (p<0.01), 75, 90, and 105 d (p<0.05) of pregnancy and at 60 (p<0.01), 45, and 105 d (p<0.05) of postnatal growth. Furthermore, based on ER genotypes, a high litter size group with genotypes AB and BB showed lower IFG-II concentration than a low litter size group with a genotype AA during pregnancy. Taken together, the results indicate that the serum IFG-II and IGFBP-3 are correlated with the litter size in pigs.

Simulation Model of Dual-Species Biofilm Growth in Hydrodynamic Flow (유체 흐름 안에서 두 종의 생물막 성장 시뮬레이션 모델)

  • Jeon, Won-Ju;Lee, Sang-Hee
    • Journal of the Korea Society for Simulation
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    • v.20 no.1
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    • pp.97-105
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    • 2011
  • In rivers and streams, biofilms are thin layers of greenish-brown slime attached to rocks, plants, and other surfaces. Biofilms play key roles in primary production and cycling of nutrients, water quality remediation, suspended sediment removal, and energy flow to higher trophic levels. In the present study, we developed a two-dimensional cellular automata model to simulate mixed biofilms of toxin-sensitive and toxin-producing species in hydrodynamic flow. The flow was generated by a stochastic process for uniform flow and by using the Navier-Stokes equation for non-uniform flow. Minimized local rules governing reproduction and mortality of the species were executed in the self-organizing processes to elucidate interactions between toxin-producing and toxin-sensitive species in competition over nutrients. We briefly discuss the morphology of the simulated biofilm under different flow conditions.