• Title/Summary/Keyword: Roles in reproduction

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Expression of Vimentin Intermediate Filament for Vascular Development in Olive Flounder (Paralichthys olivaceus)

  • Yang, Hyun;Lee, Jang-Wook;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon;Lee, Jeong-Ho
    • Development and Reproduction
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    • v.18 no.2
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    • pp.107-115
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    • 2014
  • Cardiovascular system is the primary organ to develop and reach a functional state, which underscores the essential role of the vasculature in the developing embryo. The vasculature is a highly specialized organ that functions in a number of key physiological works including the carrying of oxygen and nutrients to tissues. It is closely involved in the formation of heart, and hence it is essential for survival during the hatching period. The expression of genes involved during vascular development in the olive flounder (Paralichthys olivaceus) in the days after hatching is not fully understood. Therefore, we examined the expression patterns of genes activated during the development of flounder. Microscopic observations showed that formation of blood vessels is related to the expression of the vimentin gene. Also, the temporal expression patterns of this vimentin-like gene in the developmental stages and in the normal tissues of olive flounder. The purpose of this study was to examine the expression patterns of vimentin in normal tissues of the olive flounder and during the development of the vascular system in newly hatched olive flounders and HIF-1 plays a vital role in the formation of blood vessels during development. Vimentin expression was strong at the beginning of the development of blood vessels, and was present throughout all developmental stages. Our findings have important implications with respect to the roles of vimentin and HIF-1 in the development and evolution of the first blood vessels in olive flounder. Further studies are required to elucidate the vimentin-mediated hypoxic response signal transduction and to decipher the functional role of vimentin in developmental stages.

Ophiostomatoid Fungi in Pine Wilt Disease and Oak Wilt Disease in Korea

  • Kim, Seong Hwan
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.41-41
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    • 2014
  • Pinewood nematode (PWN, Bursaphelenchus xylophilus) is a serious pathogenic worm that quickly dry pine trees to death. Recently, PWN has been devastating huge amounts of conifer trees in Korea. As a first step to explore the association and ecological roles of fungi in PWN life cycle in Korea, in this study we first isolated and indentified fungi from PWN-infested Korean pine and Japanese black pine wood sampled in Jinju, Sacheon, Pocheon, Chuncheon, Gwangju, and Hoengseong in Korea. A total of 144 fungal isolates were obtained from Japanese black pine wood and 264 fungal isolates from Korean pine wood. Their morphology and nucleotide sequences of the ITS rDNA and ♌-tubulin gene were examined for species identification. Ophiostoma ips, Botrytis anthophila, Penicillium sp., Hypocrea lixii, Trichoderma atroviride, O. galeiforme, Fusarium proliferatum were identified from Japanese black pine wood. Leptographium koreanum, L. pini-densiflorae, Ophiostoma ips, Penicillium raistrick, Trichoderma sp. were isolated from Korean pine wood. O. ips and L. koreanum were the major species on the two different PWN-infected pine tree. The cultivation of PWN on fungal mat of the identified species did some enhance PWN reproduction. The ambrosia beetle, Platypus koryoensis, is a serious pest of oak trees in Korea. In this study we investigated filamentous fungi present in the body of the beetle. Fourteen genera of filamentous fungi belonging to Ascomycota and Basidiomycota were isolated. All the obtained genera were isolated in the mitosporic state. The identified fungi were classified in 11 distinct orders including the Ascomycota (Eurotiales, Hypocreales, Microascales, Ophiostomatales, Pleosporales, and Sordiales) and Basidiomycota (Agaricales, Corticiales, Polyporales, and Russulales Xylariales). Within Ascomycota, 13 species were found. Meanwhile five species were found within Basidiomycota. The results showed the presence of diverse fungi in P. koryoensis. Among the isolated fungi, some were able to produce wood degrading enzymes. Further fungal isolation was performed with P. koryoensis infested Quercus mongolica trees sampled at Kumdan mountain in Hanam-Si, Gyeonggi province from June of 2009 to June of 2010. Penicillin spp. and Trichoderma spp. were the major species of mold fungi group. Pichia guilliermondii was the major species of mold yeast group. Raffaelea quercus-mongolicae was also isolated, but its isolation frequency was not high. Other species identified were Ambrosiella xylebori, Fusarium solani, Cryphonectria nitschke, Chaetomium globosum, and Gliocladium viride, Candida kashinagacola, C. maritima, C. vanderkliftii, Saccharomycopsis crataegensis.

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Expression Analysis of Matrix Metalloproteinases and Tissue Inhibitor of Matrix Metalloproteinases from In Vitro Maturation Oocytes Complexes in Porcine (돼지 체외성숙난자에서 MMPs와 TIMPs의 발현 분석)

  • Kim, Sang-Hwan;Kang, Hyun-Ah;Kim, Dae-Seung;Lee, Myeong-Seop;Seo, Kang-Suk;Min, Kwan-Sik;Yoon, Jong-Taek
    • Reproductive and Developmental Biology
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    • v.34 no.1
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    • pp.55-62
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    • 2010
  • Matrix metalloproteinases (MMP) play important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, oocytes development and ovulation. In an attempt to investigate the effect of MMP activation in development cumulus-oocytes complexes, we examined the localization and expression of MMP, and monitored MMP expression profile. Cumulus-oocytes complexes were collected and matured in vitro for 24 hr, 36 hr and 48 hr. A mRNA expression of MMP-2, MMP-9, TIMP-2 and TIMP-3 was detected in all culture medium regardless of CC, DC and CDCs. Activity of MMP-2 in the DC progressively was increased from 24 hr to 48 hr. But MMP-9 was not detected in all culture medium. The localization of MMP-2 was also measured by immunohistochemistry analysis. The MMP-2 and TIMP-2 was detected in cumulus cell and oocyte zone pellucida. Expression of MMP-2 protein in the COCs was progressively increased from 24 hr to 48 hr. However, MMP-9 protein was progressively decreased from 24 hr to 48 hr. And TIMP-2 protein was most highly expressed in the CDCs 36 hr. Expression of TIMP-3 protein in the CDCs was progressively increased from 24 hr to 48 hr. In conclusion, these results suggest that MMP-2 plays a role in maintaining normal maturation and development by controlling the ECM inhibitor concentration on cumulus cell and oocytes.

Leukemia inhibitory factor and its receptor: expression and regulation in the porcine endometrium throughout the estrous cycle and pregnancy

  • Yoo, Inkyu;Chae, Soogil;Han, Jisoo;Lee, Soohyung;Kim, Hyun Jong;Ka, Hakhyun
    • Asian-Australasian Journal of Animal Sciences
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    • v.32 no.2
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    • pp.192-200
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    • 2019
  • Objective: Leukemia inhibitory factor (LIF) binds to a heterodimeric receptor composed of LIF receptor (LIFR) and glycoprotein 130 (GP130) to transmit signals into the cell. LIF plays an important role in reproduction by regulating immune response, decidualization, and implantation in several species. However, the expression of LIF and LIFR in the endometrium throughout the estrous cycle and pregnancy in pigs is not fully understood. Methods: We analyzed the expression of LIF and LIFR in the endometrium on days 0 (estrus), 3, 6, 9, 12, 15, and 18 of the estrous cycle, and days 12, 15, 30, 60, 90, and 114 of pregnancy, in conceptuses on days 12 and 15, and in chorioallantoic tissues on days 30, 60, 90, and 114 of pregnancy in pigs. We also determined the effects of estrogen and progesterone on the expression of LIF and LIFR in endometrial tissues. Results: The expression of LIF increased in the endometrium during the late diestrus phase of the estrous cycle and during mid- to late- pregnancy, while the expression of LIFR increased during early pregnancy. The expression of LIF was induced by increasing doses of estrogen, whereas the expression of LIFR was induced by increasing doses of progesterone. Conclusion: These results indicate that the expression of LIF and its receptor LIFR in the endometrium is regulated in a stage-specific manner during the estrous cycle and pregnancy, suggesting that LIF and its receptor signaling system may play critical roles in regulating endometrial function in pigs.

Molecular and Cellular Analyses of NCP, a Nuclear and Centrosomal Protein in Mouse Gametes and Early Embryos

  • Oh, Hwa-Soon;Youn, Hong-Hee;Lee, Kwang-Hee;Son, Chae-Ick;Lee, Sang-Ho
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.97-97
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    • 2003
  • For many animals the centrosome consists of a pair of centrioles and surrounding pericentriolar materials (PCMs). PCMs have been known to play roles during cell division. It is known that centrioles are necessary to assemble centrosomal components. However, many types of oocytes undergo meiosis without centrioles. It is known that in nonmurine mammalian species, the sperm introduces an intact proximal centriole unlike sea urchin where two centrioles are introduced. In case of mouse sperm, the presence of centrosome is not clear In this study, a monoclonal antibody was developed to investigate centrosome during mouse germ cell and early embryo development. Results of immunostaining and Western blotting in CHO cells suggest that the monoclonal antibody recognizes a nuclear and centrosomal protein, thus called NCP. The NCP monoclonal antibody was used to screen a cDNA expression library prepared from 12.5 mouse brain to isolate NCP gene. Nucleotide size of NCP gene obtained from immunoscreening was about 5.5kb. It is determined that the NCP may be closely related with pericentriolar material -1 gene (Pcm-1) from the result of sequencing analysis. The molecular weight, 66kDa, calculated by known DNA sequence in database is consistent with that of detected from Western blotting using CHO cell lysates. Therefore, it is assumed that NCP may be alternative splicing form of Pcm-1 of which molecular weight is 228kDa. In mouse oocytes, NCP was distributed in nucleus as in CHO cells. It was shown that the NCP was localized around neck region, probably the centrosome in mouse neck region. Interestingly, dramatic change in distribution of NCP was also shown in male germ cell development. Finally, we observed the cellular distribution of NCP during early embryo development. NCP was detected in nucleus as well as centrosome foci. It is suggested that the centrioles reassembly we occurring in blastocysts and then affects the distribution of NCP.

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Relationship between Sloan-Kettering Virus Expression and Granulosa Cells of Atretic Follicles in the Rat Ovary

  • Kim, Hyun;Matsuwaki, Takashi;Yamanouchi, Keitaro;Nishihara, Masugi;Kim, Sung-Woo;Ko, Yeoung-Gyu;Yang, Boh-Suk
    • Reproductive and Developmental Biology
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    • v.35 no.3
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    • pp.341-348
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    • 2011
  • Sloan-Kettering virus gene product of a cellular protooncogene c-Ski is an unique nuclear pro-oncoprotein and belongs to the Ski/Sno proto-oncogene family. Ski plays multiple roles in a variety of cell types, it can induce both oncogenic transformation and terminal muscle differentiation when expressed at high levels. The aim of the present study was to locate Ski protein in rat ovaries in order to predict the possible involvement of Ski in follicular development and atresia. First, expression of c-Ski mRNA in the ovaries of adult female rats was confirmed by RT-PCR. Then, ovaries obtained on the day of estrus were subjected to immunohistochemical analysis for Ski and proliferating cell nuclear antigen (PCNA) in combination with terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL). Ski was expressed in granulosa cells that were positive for TUNEL, but negative for PCNA, regardless of the shape and size of follicles. Expression of Ski in TUNEL-positive granulosa cells, but not in PCNA-positive granulosa cells, was also verified in immature hypophysectomized rats having a single generation of developing and atretic follicles by treatment with equine chorionic gonadotropin (eCG). These results indicate that Ski is profoundly expressed in the granulosa cells of atretic follicles, but not in growing follicles, and suggest that Ski plays a role in apoptosis of granulosa cells during follicular atresia.

Comparative Analysis of a Putative HLH Transcription Factor Responsible for Conidiation in Aspergillus Species

  • Abdo Elgabbar, Mohammed A.;Jun, Sang-Cheol;Kim, Jong-Hwa;Jahng, Kwang-Yeop;Han, Dong Min;Han, Kap-Hoon
    • 한국균학회소식:학술대회논문집
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    • 2015.11a
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    • pp.28-28
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    • 2015
  • Asexual reproduction or conidiation in aspergilli is a primary mean to produce their progenies that is environmentally and genetically controlled tightly. Previously, intensive researches in the model fungus Aspergillus nidulans disclosed some genes playing important roles in asexual and sexual development. Among them, one gene encoding a putative helix-loop-helix (HLH) transcription factor, named ndrA, has been isolated and characterized as a downstream regulator of developmental master regulator NsdD. By using comparative genome search of A. niduans NdrA protein, its orthologues have been identified in A. fumigatus and A. flavus, respectively (AfudrnA and AfldrnA). Deletion of the ndrA genes in both Aspergillus species made them unable to produce the conidia yet abundant production of sclerotia in A. flavus. Complementation of ndrA deletion strains by intact ndrA ORFs has restored the conidiation as in the control strains. In A. fumigatus, ndrA deletion also resulted in loss of conidiation phenotype. Northern analyses showed that the ndrA genes in both Aspergillus species are highly expressed at the early stage of the conidiation. Interestingly, the ndrA genes were found to be necessary for the proper expression of brlA genes. Antifungal sensitivity test revealed that the ndrA genes might be responsible for the sensitivity or resistance to some antifungal agents. However, ndrA deletion did not greatly influence the growth in both strains. And the A. flavus ndrA gene did not affect the aflatoxin production. Taken together, ndrA genes in Aspergillus species could be an important positive regulator of conidiation under the regulation of the nsdD gene yet upstream of the brlA gene.

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MoJMJ1, Encoding a Histone Demethylase Containing JmjC Domain, Is Required for Pathogenic Development of the Rice Blast Fungus, Magnaporthe oryzae

  • Huh, Aram;Dubey, Akanksha;Kim, Seongbeom;Jeon, Junhyun;Lee, Yong-Hwan
    • The Plant Pathology Journal
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    • v.33 no.2
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    • pp.193-205
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    • 2017
  • Histone methylation plays important roles in regulating chromatin dynamics and transcription in eukaryotes. Implication of histone modifications in fungal pathogenesis is, however, beginning to emerge. Here, we report identification and functional analysis of a putative JmjC-domain-containing histone demethylase in Magnaporthe oryzae. Through bioinformatics analysis, we identified seven genes, which encode putative histone demethylases containing JmjC domain. Deletion of one gene, MoJMJ1, belonging to JARID group, resulted in defects in vegetative growth, asexual reproduction, appressorium formation as well as invasive growth in the fungus. Western blot analysis showed that global H3K4me3 level increased in the deletion mutant, compared to wild-type strain, indicating histone demethylase activity of MoJMJ1. Introduction of MoJMJ1 gene into ${\Delta}Mojmj1$ restored defects in pre-penetration developments including appressorium formation, indicating the importance of histone demethylation through MoJMJ1 during infection-specific morphogenesis. However, defects in penetration and invasive growth were not complemented. We discuss such incomplete complementation in detail here. Our work on MoJMJ1 provides insights into H3K4me3-mediated regulation of infection-specific development in the plant pathogenic fungus.

Role of the insulin-like growth factor system in gonad sexual maturation in Pacific oyster Crassostrea gigas

  • Moon, Ji-Sung;Choi, Youn Hee
    • Fisheries and Aquatic Sciences
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    • v.23 no.2
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    • pp.3.1-3.8
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    • 2020
  • Background: The IGF system plays important roles in controlling growth, development, reproduction, and aging of organisms. Methods: To estimate maturation of the Pacific oyster Crassostrea gigas, we investigated the expression of insulin-like growth factor (IGF) system components and sex-specific genes. To determine the role of the IGF system in the growth and spawning period of female and male oysters, we examined mRNA expression levels of the C. gigas insulin receptor-related receptor (CIR), IGF binding protein complex acid labile subunit (IGFBP_ALS), and molluscan insulin-related peptide (MIP), as well as those of vitellogenin (Vg) and receptor-type guanylate cyclase (Gyc76C) in gonads of C. gigas collected between April and October, when sex can be determined visually in this species. Results: We found that MIP, IGFBP_ALS, and CIR mRNA expression levels were dependent on sex and month and were greater in males than in females. CIR and Vg mRNA expression levels were very similar among females, whereas IGF system components and Gyc76C were very similarly expressed among males. The highest expression values were observed in May, when oysters are mature; CIR and Vg mRNA expression levels were highest in females, and those of MIP, IGFBP_ALS, CIR, and Gyc76C were highest in males. Interestingly, we observed a 1:1 proportion of females to males during this period. Conclusion: Our results suggest that IGF system components, as well as Vg and Gyc76C, are associated with sexual maturation in C. gigas.

Effects of BSA, PVA, Gonadotropins and Follicle Shell on In Vitro Maturation and In Vitro Fertilization of Porcine Oocytes

  • Cong, Pei-Qing;Song, Eun-Sook;Kim, Eui-Sook;Li, Zhao-Hua;Zhang, Yong-Hua;Yi, Young-Joo;Park, Chang-Sik
    • Reproductive and Developmental Biology
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    • v.31 no.2
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    • pp.61-69
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    • 2007
  • This study was designed to evaluate effects of BSA, PVA, gonadotropins and follicle shell during IVM of porcine oocytes and subsequent development to the blastocyst stage after IVF. Cumulus oocyte complexes (COCs) were cultured in TCM-199 media containing 4 mg/ml BSA and 1 mg/ml PVA during IVM for 44 hr. To compare the effect of gonadotropins on oocyte maturation, COCs were cultured with FSH+LH, FSH, LH and FSH-LH-free media during IVM. respectively. Also, different number of follicle shells (0, 2, 4 and 6) was used to examine whether the presence of follicle shell in culture medium affects oocyte maturation. The percentages of fertilization and blastocyst formation, respectively, were higher in the medium containing the PVA (49.0 and 17.9%) than those containing the BSA (40.0 and 12.2%). Significantly higher rates of Mil oocytes were in the presence of FSH+LH and FSH (88.6 and 85.1 %) compared to other treatments (64.0 and 53.4% at LH and FSH-LH-free media). Co-culture with inverted follicle shells in 2 ml maturation medium enhanced the developmental competence of porcine oocytes. In conclusion, PVA could be used as a macromolecules instead of BSA, and FSH and follicle shell played important roles in maturation of porcine oocytes.