• Title/Summary/Keyword: 웅성생식세포

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Effect of ${\beta}-Mercaptoethanol$ Supplement during In Vitro Maturation on IVM, IVF and Glutathione Level in Bovine Oocytes (소 미성숙 난포란의 체외성숙시 ${\beta}-Mercaptoethanol$의 첨가가 체외성숙, 체외수정 및 Glutathione 수준에 미치는 영향)

  • Oh, Shin-Ae;Kim, Chang-Keun;Chung, Yung-Chai;Pang, Myung-Geol
    • Development and Reproduction
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    • v.10 no.4
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    • pp.239-245
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    • 2006
  • Experiments were conducted to determine the effects of beta-mercaptoethanol(${\beta}-ME$) supplements to the maturation medium on in vitro fertilization(IVF) and intracellular glutathione(GSH) concentration. Bovine cumulus-intact oocytes were matured in TCM-199 medium containing FBS, hormonal supplements, and ${\beta}-ME$(0, 25 and $50\;{\mu}M$) for 12h and 24 h. After culture, cumulus-free matured oocytes were co-incubated with frozen-thawed spermatozoa for 24h. Maturation rate increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant differences among treatment groups. Also, increases(p<0.05) in intracellular GSH concentration before and after fertilization were observed in $50\;{\mu}M\;{\beta}-ME$ supplements to the maturation medium. Male pronuclear formations after IVF was increased(p<0.05) in ${\beta}-ME$ treatment group, but no significant difference among treatment groups. In conclusion, supplementing ${\beta}-ME$ into the maturation medium increased maturation rates, fertilization rates, and intracellular GSH concentrations.

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Aberrant Microtubule Assembly and Chromatin Configuration of Homan Oocytes Which Failed to Complete Fertilization Following In Vitro Fertilization and Intracytoplasmic Sperm Injection (일반적 수정과 세포질내 정자주입법에 의해 수정에 실패한 인간난자의 미세소관과 염색체의 형태이상)

  • Chung, H. M.;Kim, N. H.;Kim, J. W.;J. M. Lim;Park, C.;J. J. Ko;K. Y. Cha;Kim, J. M.;K. S. Chung
    • Korean Journal of Animal Reproduction
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    • v.24 no.2
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    • pp.143-154
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    • 2000
  • Most eggs initiated the fertilization processes but arrested at specific stages. The stages included failure of the oocyte to exit from the meiotic metaphase-II with or without sperm penetration, failure of appropriate sperm aster formation, inability to form proper male and female pronuclei, failure of suitable pronuclear apposition, and failure to form proper number of either male or female pronuclei. Various images of defective microtubule organization and chromatin configuration during IVF and ICSI procedures were observed. We discussed the data with previous research results during normal fertilization in humans and other mammals. In conclusion, various aberrant patterns in microtubule assembly and chromatin configuration, which were assessed in the present study, could be used as criteria to improve assisted reproductive technology in clinics. However, further cellular and molecular characterization is needed to clarify these aberrant patterns of cytoskeletal assembly.

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DDT Reduced Testosterone and Aromatase Activity Via ER Receptor in Leydig Cell (DDT의 Aromatase 증가에 의한 Testosterone 감소효과)

  • Lee, Kyung-Jin;Wui, Seong-Uk;Jin Heo;Kim, Sun-Hee;Jeong, Ji-Yeon;Lee, Jong-Bin
    • Environmental Analysis Health and Toxicology
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    • v.18 no.2
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    • pp.95-100
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    • 2003
  • Dichlorodiphenyltrichloroethane (DDT), is a widespread environmental pollutant. In this study, we investigated the effect of DDT on testosterone production through aromatase and investigated its molecular mechanism in testicular leydig cell, R2C. We investigated that the effects of DDT on testosterone production and its effects on aromatase activity in R2C cell by radio immunoassay (RIA). As the results, the potent leyding cell activator LH increased testosterone production compared to the control. DDT exposure significantly decreased testosterone production in R2C cell and DDT alone affected T reduction in a dose-dependent manner in R2C cell slightly. In addition, DDT was found to increase aromatase activity in R2C cell in a dose dependent manner. In order to assess whether the suppressive effects of DDT on LH-inducible testosterone production might be influenced by the ER, ICI 182.780, a pure antiestrogen, was used, and it was found that these inhibitory effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the suppressive effects of DDT. Furthermore, the inducible effects of DDT on aromatase might be influenced by the ER, ICI 182.780 was used, and it was found that these enhancing effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the inducible effects of DDT. Our results indicated that DDT inhibition of LH-inducible testosterone production in R2C is mediated through aromatase. However, the precise mechanisms by which DDT enhance in leyding cell remains unknown. The current study suggests the possibility that DDT might act as a modulator aromatase gene transcription.

Effects of Di-(2-ethylhexyl) phthalate (DEHP) on Ultrastructure of Rat Seminal Vesicle (Di-(2-ethylhexyl)phthalate (DEHP)가 흰쥐 저정낭의 미세구조에 미치는 영향)

  • Kil, Young-Chun;Kim, Wan-Jong;Shin, Kil-Sang
    • Applied Microscopy
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    • v.30 no.1
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    • pp.73-80
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    • 2000
  • Di-(2-ethylhexyl)phthalate (DEHP) has been known as one of endocrine disruptors. The present study was carried out to investigate the alterations of fine structure in rat seminal vesicle after oral intubation of DEHP in dosages of 1g/kg/day, 2g/kg/day or 3g/kg/day respectively in 0.5 ml of corn oil for If days. In rats treated with DEHP for 15 days, seminal vesicle exhibited extensive histological alterations compared to those observed in control groups. The size of the seminal vesicle and the mucosal folds decreased, but the lamina propria was considerably thickened. The ultrastructural changes of epithelial cells in seminal vesicle of rat treated with DEHP were characterized by the high nuclear/cytoplasmic ratio and the increased beterochromatin within irregular nuclear envelope. And also, the rough endoplasmic reticulum, Golgi complex and secretory vesicles were poorly developed. In conclusion, DEHP caused the ultrastructural and functional alterations of seminal vesicle in rats dose-dependently. It is suggested that these detrimental effects of DEHP on seminal vesicle are derived from the decrease level of testosterone.

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Effects of 3,5,3'-triiodo-L-thyronine ($T_3$) on Sex Steroid Levels and Gonadal Development in Black Porgy, Acanthopagrus schlegeli (감성돔, Acanthopagrus schlegeli의 성 스테로이드 및 생식소 발달에 미치는 3,5,3'-triiodo-L-thyronine ($T_3$)의 영향)

  • Min, Byung-Hwa;Noh, Gyoung-Ane;Jeong, Min-Hwan;Chang, Young-Jin
    • Development and Reproduction
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    • v.9 no.1
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    • pp.15-22
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    • 2005
  • The objective of the present study was to investigate changes of sex steroid(testosterone: T and $estradiol-17{\beta}:\;E_2$), cortisol levels and gonadal development following $T_3$ treatment to protandrous black porgy, Acanthopagrus schlegeli. Exogenous $T_3$ was found to significantly stimulate the increase of T levels in plasma of black porgy after 60 days of treatment. However no effects of $T_3$ on $E_2$ levels and oocyte size were found. $T_3$ treatment resulted in stimulated spermatogenesis and testicular development in gonad and prolonged spermiation. Also, the levels of cortisol were significantly increased in the fish treated with $T_3$ as compared to control fish at 60 days. The results showed that exogenous $T_3$ had direct effect on the release of T and cortisol, thus $T_3$ seems to play, either directly or indirectly, an important role in the testis development of functional male black porgy.

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Effect of Phyto-Extract Fermented Mixture (MP119) on the Sexual Functions and on the Toxicities of Cadmium (식물추출복합발효물(MP119)이 성기능에 미치는 영향 및 카드뮴 독성에 대한 효과)

  • Jang, Young-Sun;Jeong, Jong-Moon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.38 no.12
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    • pp.1724-1731
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    • 2009
  • This study was aimed to investigate the effect of phyto-extract fermented mixture (MP119) on the male sexual functions. The MP119 was evaluated for anti-impotency and anti-hypertensive effects via ACE (angiotensin converting enzyme) or PDE (phosphodiesterase) inhibition assay. $IC_{50}$ values of MP119 against ACE and PDE were 241.3${\pm}$35.5 ppm and 372.2${\pm}$33.8 ppm, respectively. To investigate the effect of testosterone expression by MP119, we performed cell media test using mouse Leydig-derived TM3 cells. Production of testosterone in TM3 cell was increased by MP119. Also, NO (nitric oxide) production of HUVEC (human umbilical vein endothelial cell) was increased when MP119 was added to the cultures. Forty male ICR mice were divided into 4 groups. MP119 was orally intubated for 7 days to group 1 and 3, and same volume of vehicle to group 2 and 4 as controls. After that, group 3 and 4 were intraperitoneally injected cadmium chloride at a single dose of 2 mg/kg. On the 8th experimental day, weights of testis, epididymis and seminal vesicle, number of sperm, concentrations of serum testosterone and cGMP were determined. The number of sperm, the concentrations of testosterone and cGMP were significantly increased in two experimental groups (group 1, 3). These results suggest that MP119 enhanced the sexual function of male mice, and could protect the sexual organs from the cadmium chloride as one of the endocrine disrupters.

Gonadal Development and the Effects of $17^{\alpha}$-methyltestosterone on Sex Inversion of the Red Spothed Grouper, Epinephelus akaara (붉바리, Epinephelus akaara의 생식소 발달과 $17^{\alpha}$-methyltestosterone 처리 효과)

  • Hwang, Sung-il;Lee, Young-Don;Song, Choon-Bok;Rho, Sum
    • Journal of Aquaculture
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    • v.11 no.2
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    • pp.173-182
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    • 1998
  • The study has been conducted to understand gonadal development and the effects of $17^{\alpha}$-methyltestosterone on sex inversion of the red spotted grouper, Epinephelus akaara. Fish were collected from Deukyand bay in the southern coast of Korea in August, 1996 and then they had been cultivated at the indoor tank until August, 1997. Gonad somatic index (GSI) in the females of both treated and control group began to increase from February when water temperature was rainse again, and reached the maximum value in August, whereas it had decreased from September adn thereafter maintained relatively low value until January. Unlike females, GSI in the male or intersex of treated groups decreased after June. Hepatosomatic index (HSI) of the control group tended to show the relatively low around Autumn, whereas it showed relatively highr value in April and June when the ovary was in the growing stage. Although the treated groups showed relatively higher value of the HSI than the control, hte paterns in monthly variation of HSI were similar to the control. Sexual change of the female grouper to the male was attempted by acceleration with oral administration of $17^{\alpha}$-methyltestosterone at the dose of 0.2 and 0.5mg/kg fish for 120days. Transitional hermaphroditic gonads were observed from the various size of groupers ranging 21.0 to 36.1 cm in total length, while the functional males could be induced from th individuals of 28.8 to 33.5cm in total length. This result indicated that larger groupers than 30cm in total length should be used for sex inversion to maleness with $17^{\alpha}$-methyltestosterone.

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Applications of PCR and PRINS for the Sexing in Bovine Preimplantation Embryos (착상전 소 초기배의 성판정을 위한 PCR과 PRINS의 적용)

  • Seo, Seung-Woon;Lee, Hong-Jun;Kim, Ki-Dong;Park, Sung-Soo;Lee, Sang-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.23 no.3
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    • pp.341-349
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    • 1996
  • 초기배의 성판정은 대상가축의 성을 선발하는 수단으로써 뿐만아니라 인간의 유전적 질병의 조기진단법으로서 매우 가치가 크다. 체외수정 소 초기배의 성을 결정하기 위해 PCR과 PRINS를 이용하였으며 성판정에 이용된 8 세포${\sim)$배반포기 초기배는 체외수정후 난관상피세포와의 공배양에 의해 생산되었다. 초기배 의 DNA는 $200{\mu}g/ml$ proteinase K가 함유된 PCR lysis buffer에 하나의 초기배를 부유한 후 $50^{\circ}C$에서 1시간동안 처리하여 준비하였다. 중기 염색체 spreads는 초기배를 nocodazole로 처리한 후 air-drying 방법을 이용하여 준비하였다. 가능한 false positive signals을 배제하기 위해 소특이 및 Y 염색체 특이 primers를 이용하여 PCR을 수행한 결과, 웅성 초기배에서는 두 개의 증폭산물 (소특이 및 Y 염색체 특이)이 합성된 반면 자성 초기배에서는 하나의 증폭산물만 합성되었다. 한편 중기염색체상의 Y 염색체를 동정하기 위해 FISH와 PRINS를 수행한 결과, FISH에서보다 PRINS에서 더 강한 Y 염색체 특이 형광 signals이 탐지되었다. 이러한 결과는 PCR에 의한 체외생산 소 초기배의 신속정확하고 효율적인 성판정이 가능함을 보여주었다. 또한 PRINS를 통해 PCR 에 이용된 Y 특이 probe의 신뢰성이 염색체 수준에서 확인되었다.

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Immunohistochemical study on the antigenicity of each organ structure of Clonorchis sinensis (간흡충 충체의 부위별 항원성에 대한 면역조직화학적 연구)

  • Jin Kim;Jong-Yil Chai;Weon-Gyu Kho;Kyu-Hyuk Cho;Soon-Hyung Lee
    • Parasites, Hosts and Diseases
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    • v.29 no.1
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    • pp.21-30
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    • 1991
  • An immunohistochemical study was performed to demonstrate comparative antigenicity of each body structure of the liver fluke, Clonorchis sinensis, such as the digestive tract, reproductive organs, excretory system, tegument and suckers. Indirect immunoperoxidase technique was applied, rising formalin-fixed and paraffin-embedded sections of C. sinensis as the antigen. Pooled cat sera obtained 10 weeks after an experimental infection with C. sinensis and peroxidase-conjugated goat anti-cat IgG were used as the primary and secondary antibodies, respectively. The intensity of immunohistochemical stain was very sensitive upon the titers of the primary and secondary antibodies, and their optimum dilutions were found to be 1:1,000∼1:2,000 and 1:1,000, respectively. The intestinal epithelial cells, intestinal content and excretory bladder showed strong positive coloring reactions even at lower titer (1 : 2,000) of the primary antibody, whereas the uterine wall and eggs, vitelline glands, and male reproductive organs showed only weak positive reactions despite an increase in the antibody titer (1:1,000). On the other hand, the suckers, tegument, subtegumental cells and other parenchyme portions did not reveal any positive immunoperoxidase reaction at the same antibody titers. From the above results, it is highly suggested that the most potent antigenicity of C. sinensis occur from their excretory-secretory substances originated from the digestive and excretory organs.

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Immunohistochemieal study on the antigenicity of body compartments of Payugonimus westermani (폐흡충 충체 부위별 항원성에 대한 면역 조직화학적 연구)

  • Lee, Sun-Hyeong;Seong, Suk-Hwan;Chae, Jong-Il
    • Parasites, Hosts and Diseases
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    • v.27 no.2
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    • pp.109-118
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    • 1989
  • Production of circulating specific antibodies to the lung fluke (Paragenimus westermani) by its host is well known and used in various kinds of immunodiagnostic methods, However, it has not been well documented which compartments (or structures) of the lung fluke are most responsible for the production of specific antibodies. The present immunohistochemical study was undertaken to demonstrate the antigenicity of each body compartment of p. westermani such as suckers, tegument, spines, vitelline glands, intestine, reproductive organs(male and female), and eggs. Indiret immunoperoxidase(IP) stain technique was applied, using formalin-fked, paraffin- embedded lung tissues of P westermani-infected cats sectioned in 4 Um thickness as the antigen and cat antisera (11~20 weeks of infection) as the primary antibody. Peroxidase-conjugated goat anti-cat IgG was used as the secondary antibody and diaminobensidine(DAB) as the coloring agent. Strong yellow or yellowish brown staining was regarded positive. The primary and secondary antibody dilutions were made at 1 : 500~1 : 2, 000 and 1 : 200~1 : 500 respectively, and IP stain was repeated 10 times for each dilution. A consistent result obtained was that the intestinal epithelial border, intestinal content, vitelline glands, and eggs scattered around the worm capsule showed strong positive staining, while uterine eggs and some parenchymal portions showed weak positive reaction. On the other hand, the suckers, tegument, spines, subtegumental cells, cytoplasm of intestinal epithelial cells, male reproductive organs, and ovary revealed negative staining. The body compartments showing higher antigenicity were, in the decreasing order, the intestinal epithelial border, intestinal content, eggs in the worm capsule, vitelline glands, uterine eggs, and parenchymatous portions. The intestinal epithelial border and luminal contents revealed positive staining even at a few concentration of 1 : 4, 000 primary antibody(secondary ab., 1 : 200) whereas the parenchymatous portion showed positive reaction only at higher concentrations than 1'500 (secondary ab., 1 : 200). The results suggest that the specific antibody responses of the host to p. westermani occur most strongly upon the excretes from the intestinal epithelium of the worm and e99s Produced around the worm capsule,

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