• Title/Summary/Keyword: Reproductive system

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Development of Artificial Insemination Techniques with a Minimum Numbers of Insemination Spermatozoa using Laparoscopy

  • Lee, J. H.;Park, S. J.;I. S. Ryu;G. Y. Chung;Park, S. H.;D. Y. Ji;Kim, C. K.;S. H. Baek
    • Proceedings of the KSAR Conference
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    • 2004.06a
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    • pp.250-250
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    • 2004
  • This study was conducted to investigate the effect of the number of spermatozoa and insemination section(field) of reproductive organs at artificial insemination using laparoscopy(Fiber optic laparoscopic system, Good-Gene Co., Korea) in deer(Elk) and cattle. Twenty six elk does and fifteen cows were inserted CIDR into virginia during 12∼14 days for synchronization of estrus. (omitted)

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Generation of Transgenic Chickens Regulating hEPO Gene Expression (hEPO 유전자의 발현이 조절되는 형질전환 닭의 생산)

  • Koo, Bon-Chul;Kwon, Mo-Sun;Kim, Te-Oan
    • Reproductive and Developmental Biology
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    • v.34 no.3
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    • pp.193-199
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    • 2010
  • We report here the production of transgenic chickens that can regulate human erythropoietin (hEPO) gene expression. The glycoprotein hormone hEPO is an essential for viability and growth of the erythrocytic progenitors. Retrovirus vector system used in this study has two features including tetracycline-controllable promoter and woodchuck hepatitis virus posttranscriptional regulator element (WPRE). The former is for to reduce the possibility of physiological disturbance due to constitutional and unregulated expression of hEPO gene in the transgenic chicken. The latter is for maximum expression of the foreign gene when we turn-on the gene expression. A replication-defective Moloney murine leukemia virus (MoMLV)-based vectors packaged with vesicular stomatitis virus G glycoprotein (VSV-G) was injected beneath the blastoderm of non-incubated chicken embryos (stage X). Out of 325 injected eggs, 28 chicks hatched after 21 days of incubation and 16 hatched chicks were found to express the hEPO gene delivered by the vector. The biological activity of the recombinant hEPO in transgenic chicken serum was comparable to its commercially available counterpart. The recombinant hEPO in transgenic chicken serum had N- and O-linked carbohydrate simillar to that produced from in vitro cultured cells transformed with hEPO gene.

In Vitro Expression of the Recombinant hFSH Gene using Retrovirus Vector System (In Vitro에서 Retrovirus Vector System을 이용한 재조합 hFSH 유전자의 발현)

  • Min, Gyeong-Heon;Kwon, Mo-Sun;Kim, Teoan;Koo, Bon-Chul
    • Reproductive and Developmental Biology
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    • v.35 no.1
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    • pp.115-121
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    • 2011
  • hFSH is a glycoprotein secreted from anterior pituitary and consists of ${\alpha}$ and ${\beta}$ subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: ( i ) a DNA fragment containing ${\alpha}$ and ${\beta}$ genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ii) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.

Ovulated female salamander (Hynobius leechii) respond to water currents

  • Kim, Seok-Bum;Lee, Jung-Hyun;Ra, Nam-Yong;Park, Dae-Sik
    • Journal of Ecology and Environment
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    • v.33 no.3
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    • pp.217-222
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    • 2010
  • To determine whether or not the female Korean salamander, Hynobius leechii, responds to water currents and, if so, whether those responses depend on their reproductive conditions, we evaluated the responses of ovulated and oviposited females to 1-Hz water currents generated by a model salamander with and without the placement of a transparent water current blocker between the model and the test females. The ovulated females responded to water currents by turning their heads toward, approaching, and/or making physical contact with the model. When the water current blocker was in place, the number of salamanders that approached the model was reduced significantly. The approaching and touching responses of ovulated females were greater than those of oviposited females, whereas the other measurements evidenced no differences. None of the responses of the oviposited females to water currents was affected by the presence of the blocker. Our results indicate that female H. leechii responds to water currents via a mechanosensory system.

Endothelial Cells Isolated from the Bovine Corpus Luteum Synthesize Prostaglandin $F_{2{\alpha}}$ Receptor

  • Gwon, Sun-Yeong;Rhee, Ki-Jong;Lee, Seunghyung
    • Biomedical Science Letters
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    • v.19 no.3
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    • pp.261-265
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    • 2013
  • The corpus luteum is a transient endocrine gland essential for regulation of the ovarian cycle as well as for establishing and maintaining pregnancy. Prostaglandin $F_{2{\alpha}}$ (PGF) initiates functional and structural regression of the corpus luteum and therefore is an important regulator of the estrous cycle. It is a matter of debate whether the endothelial cells of the bovine corpus luteum express PGFR, the cognate receptor for PGF. Therefore, the aim of this study was to assess the expression of PGFR in bovine endothelial cells. Endothelial cells were isolated from the bovine corpus luteum of the mid-luteal stage using magnetic beads and cultured in vitro. We demonstrate that this isolation procedure generates a pure culture of endothelial cells as confirmed by synthesis of Factor VIII and lack of expression of $3{\beta}$-hydroxysteroid dehydrogenase. By RT-PCR, Western blot and immunofluorescence analyses, we further show that the cultured endothelial cells produced PGFR. This model system can be utilized to provide an experimental system to investigate the role of PGF on endothelial cells during the reproductive cycle.

Characterization and Expression Profile of CMTM3/CKLFSF3

  • Zhong, Ji;Wang, Yu;Qiu, Xiaoyan;Mo, Xiaoning;Liu, Yanan;Li, Ting;Song, Quansheng;Ma, Dalong;Han, Wenling
    • BMB Reports
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    • v.39 no.5
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    • pp.537-545
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    • 2006
  • CMTM/CKLFSF is a novel family of proteins linking chemokines and TM4SF. In humans, these proteins are encoded by nine genes, CKLF and CMTM1-8/CKLFSF1-8. Here we report the characteristics and expression profile of CMTM3/CKLFSF3. Human CMTM3/CKLFSF3 has a high sequence identity among various species and similar characteristics as its mouse and rat homologues. Established by results both of RT-PCR and Quantitative Real-time PCR, the gene is highly transcribed in testis, leukocytes and spleen. For further verification, we generated a polyclonal antibody against human CMTM3/CKLFSF3 and found that the protein is highly expressed in the testis and some cells of PBMCs. Therefore, CMTM3/CKLFSF3 is an evolutionarily conserved gene that may have important roles in the male reproductive system and immune system. Further studies are necessary to validate its functions in the two systems.

Analysis of C. elegans VIG-1 Expression

  • Shin, Kyoung-Hwa;Choi, Boram;Park, Yang-Seo;Cho, Nam Jeong
    • Molecules and Cells
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    • v.26 no.6
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    • pp.554-557
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    • 2008
  • Double-stranded RNA (dsRNA) induces gene silencing in a sequence-specific manner by a process known as RNA interference (RNAi). The RNA-induced silencing complex (RISC) is a multi-subunit ribonucleoprotein complex that plays a key role in RNAi. VIG (Vasa intronic gene) has been identified as a component of Drosophila RISC; however, the role VIG plays in regulating RNAi is poorly understood. Here, we examined the spatial and temporal expression patterns of VIG-1, the C. elegans ortholog of Drosophila VIG, using a vig-1::gfp fusion construct. This construct contains the 908-bp region immediately upstream of vig-1 gene translation initiation site. Analysis by confocal microscopy demonstrated GFP-VIG-1 expression in a number of tissues including the pharynx, body wall muscle, hypodermis, intestine, reproductive system, and nervous system at the larval and adult stages. Furthermore, western blot analysis showed that VIG-1 is present in each developmental stage examined. To investigate regulatory sequences for vig-1 gene expression, we generated constructs containing deletions in the upstream region. It was determined that the GFP expression pattern of a deletion construct (${\Delta}-908$ to -597) was generally similar to that of the non-deletion construct. In contrast, removal of a larger segment (${\Delta}-908$ to -191) resulted in the loss of GFP expression in most cell types. Collectively, these results indicate that the 406-bp upstream region (-596 to -191) contains essential regulatory sequences required for VIG-1 expression.

Mathematical Modelling for the Transmission Dynamics of HIV infection and AIDS (HIV감염과 AIDS의 전파특성에 관한 수학적 모델화)

  • Chung, Hyeng-Hwan;Joo, Seok-Min;Chung, Mun-Gyu;Lee, Kwang-Woo
    • The Transactions of the Korean Institute of Electrical Engineers A
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    • v.48 no.2
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    • pp.89-95
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    • 1999
  • This study investigates the population model of the spread of HIV/AIDS which the infection is generated by an infectious individual in a population of susceptible. A mathematical model is presented for the transmission dynamics of HIV infection within the communities of homosexual males. The pattern on the epidemic character of HIV, the causative agent of AIDS, was analysed by the mathematical model of AIDS system which is derived according to the ecological relationship between five epidemilogic states of individuals. The computer simulation was performed using real data and the following conclusions are drawn on the basis of the simulations. 1. The model structure and the algorithm described n the thesis is good. 2. In proportion to increase Ro, the population of AIDS patient increases and the time of its widespread reaches earlier. 3. The AIDS patients will be maximum between 7 and 21 years after an attack of AIDS and widespread between 10 and 20 years. 4. Considering the properties of the incubation periods, the maximum number of infected person is increased, and the attack rate is decreased.

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Expression of Maturation-Related Genes and Leptin during Sexual Maturation in the Female Goldfish: Effects of Exogenous Kisspeptin

  • Kim, Na Na;Choi, Young Jae;Oh, Sung-Yong;Choi, Cheol Young
    • Journal of Marine Life Science
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    • v.1 no.1
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    • pp.41-49
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    • 2016
  • Kisspeptin (Kiss) and its cognate receptor, kisspeptin receptor (KissR; G protein coupled receptor 54, GPR54), have recently been recognized as potent regulators of reproduction in teleosts. Additionally, leptin plays an important role in energy homeostasis and reproductive function in teleosts. The purpose of this study was to examine differences in the concentration of the hormones of the Kiss/KissR system and leptin and the expression of their underlying genes, all of which are involved in the sexual maturation of female goldfish, Carassius auratus, following treatment with Kiss. The expression levels of KissR increased after the Kiss injection. Furthermore, the peptide hormone leptin also increased after the injection (in vivo and in vitro). Additionally, the expression of GnRH and GTHs (GTHα, FSHβ, and LHβ) increased in the brain and pituitary (in vitro and in vitro). These results support the hypothesis that Kiss plays important roles in the direct regulation of the hypothalamus-pituitary-gonad axis and leptin in goldfish. Therefore, we suggest that Kiss system gene expression is correlated with energy balance and reproduction.

Expression of the Recombinant Porcine GH Gene In Vitro Using Tetracycline Inducible Expression System (In Vitro에서의 Tetracycline Inducible Expression System에 의한 재조합 돼지 성장호르몬 유전자의 발현)

  • Kwon Mo Sun;Koo Bon Chul;Kim Teoan
    • Reproductive and Developmental Biology
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    • v.29 no.1
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    • pp.49-55
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    • 2005
  • We cloned cDNA of the PGH(porcine growth hormone) gene and constructed retrovirus vector designed to express PGH gene under the regulation of CMV (cytomegalovirus) promoter. To maximize the expression, WPRE(woodchuck hepatitis virus posttranscriptional regulatory element) sequence was placed at the downstream of the PGH gene. After infection with recombinant viruses, approximately 1×10/sup 6/ PFF(porcine fetal fibroblast) cells released PGH protein into the media as much as 1,400 ng. In a subsequent experiment, a modifications of the retrovirus vector was made to express the PGH gene in a teracycline-inducible manner. In PFF cells carrying these viral vector sequences, addition of doxycycline to the media resulted in 2∼6 fold increase in PGH synthesis. In the modified retrovirus vectors, the WPRE sequence also played a role in boosting the effect of the tetracycline induction. This result indicates that our tetracycline-inducible expression system might be a promising candidate in alleviating the complicate physiological problems caused by constitutive expression of the exogenous genes in the transgenic animals.