• Title/Summary/Keyword: mouse testis

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Demethylation of CpG islands in the 5' upstream regions mediates the expression of the human testis-specific gene MAGEB16 and its mouse homolog Mageb16

  • Liu, Yunqiang;Wang, Meiling;Jiang, Siyuan;Lu, Yongjie;Tao, Dachang;Yang, Yuan;Ma, Yongxin;Zhang, Sizhong
    • BMB Reports
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    • v.47 no.2
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    • pp.86-91
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    • 2014
  • Tissue-specific gene expression is regulated by epigenetic modification involving trans-acting factors. Here, we identified that the human MAGEB16 gene and its mouse homolog, Mageb16, are only expressed in the testis. To investigate the mechanism governing their expression, the promoter methylation status of these genes was examined in different samples. Two CpG islands (CGIs) in the 5' upstream region of MAGEB16 were highly demethylated in human testes, whereas they were methylated in cells without MAGEB16 expression. Similarly, the CGI in Mageb16 was hypomethylated in mouse testes but hypermethylated in other tissues and cells without Mageb16 expression. Additionally, the expression of these genes could be activated by treatment with the demethylation agent 5'-aza-2'-deoxycytidine (5'-aza-CdR). Luciferase assays revealed that both gene promoter activities were inhibited by methylation of the CGI regions. Therefore, we propose that the testis-specific expression of MAGEB16 and Mageb16 is regulated by the methylation status of their promoter regions.

Effects of Gintonin-enriched fraction on the gene expression of six lysophosphatidic receptor subtypes

  • Lee, Rami;Lee, Byung-Hwan;Choi, Sun-Hye;Cho, Yeon-Jin;Cho, Han-Sung;Kim, Hyoung-Chun;Rhim, Hyewhon;Cho, Ik-Hyun;Rhee, Man Hee;Nah, Seung-Yeol
    • Journal of Ginseng Research
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    • v.45 no.5
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    • pp.583-590
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    • 2021
  • Background: Gintonin, isolated from ginseng, acts as a ginseng-derived lysophosphatidic acid (LPA) receptor ligand and elicits the [Ca2+]i transient through six LPA receptor subtypes (LPARSs). However, the long-term effects of gintonin-enriched fraction (GEF) on the gene expression of six LPARSs remain unknown. We examined changes in the gene expression of six LPA receptors in the mouse whole brain, heart, lungs, liver, kidneys, spleen, small intestine, colon, and testis after long-term oral GEF administration. Methods: C57BL/6 mice were divided into two groups: control vehicle and GEF (100 mg/kg, p.o.). After 21-day saline or GEF treatment, total RNA was extracted from nine mouse organs. Quantitative-real-time PCR (qRT-PCR) and western blot were performed to quantify changes in the gene and protein expression of the six LPARSs, respectively. Results: qRT-PCR analysis before GEF treatment revealed that the LPA6 RS was predominant in all organs except the small intestine. The LPA2 RS was most abundant in the small intestine. Long-term GEF administration differentially regulated the six LPARSs. Upon GEF treatment, the LPA6 RS significantly increased in the liver, small intestine, colon, and testis but decreased in the whole brain, heart, lungs, and kidneys. Western blot analysis of the LPA6 RS confirmed the differential effects of GEF on LPA6 receptor protein levels in the whole brain, liver, small intestine, and testis. Conclusion: The LPA6 receptor was predominantly expressed in all nine organs examined; long-term oral GEF administration differentially regulated LPA3, LPA4, and LPA6 receptors in the whole brain, heart, lungs, liver, kidneys, small intestine, and testis.

Assessment of Reproductive Health Risk of Polychlorinated Biphenyls by Monitoring the Expression of Claudius and Transepithelial Electrical Resistance in Mouse Sertoli Cells

  • Gye, Myung-Chan;Seiichiroh Ohsako;Lee, Ho-Joon
    • Journal of Microbiology and Biotechnology
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    • v.13 no.4
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    • pp.495-500
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    • 2003
  • Tight junctions (TJ) between adjacent Sertoli cells in testis are important for the formation of the blood testis barrier (BTB). In an effort to verify the reproductive health risk of endocrine-active chemicals (EACs), changes in the transepithelial electrical resistance (TER) and the expression of TJ genes were examined by co-planar polychlorinated biphenyl (PCB) treatment in cultured mouse Sertoli cells. Although the increase in TER of Sertoli cells was accelerated by 10 nM co-planar PCB, it was downregulated by 100 nM co-planar PCB. The expression of claudin-1 was downregulated by co-planar PCB in a concentration-dependent manner. On the contrary, the expression of claudin-1 was increased in the Sertoli cells by 10 nM co-planar PCB treatment. These results suggest that the structure and function of TJ may be targeted by co-planar PCB in Sertoli cells. Assessment of the structure and function of TJ in Sertoli cells might be useful for screening the reproductive health risk of EACs.

Centrobin/Nip2 Expression In Vivo Suggests Its Involvement in Cell Proliferation

  • Lee, Jungmin;Kim, Sunmi;Jeong, Yeontae;Rhee, Kunsoo
    • Molecules and Cells
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    • v.28 no.1
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    • pp.31-36
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    • 2009
  • Centrobin/Nip2 was initially identified as a centrosome protein that is critical for centrosome duplication and spindle assembly. In the present study, we determined the expression and subcellular localization of centrobin in selected mouse tissues. Immunoblot analysis revealed that the centrobin-specific band of 100 kDa was detected in all tissues tested but most abundantly in the thymus, spleen and testis. In the testis, centrobin was localized at the centrosomes of spermatocytes and early round spermatids, but no specific signal was detected in late round spermatids and elongated spermatids. Our results also revealed that the centrosome duplication occurs at interphase of the second meiotic division of the mouse male germ cells. The centrobin protein was more abundant in the mitotically active ovarian follicular cells and thymic cortex cells than in non-proliferating corpus luteal cells and thymic medullary cells. The expression pattern of centrobin suggests that the biological functions of centrobin are related to cell proliferation. Consistent with the proposal, we observed reduction of the centrobin levels when NIH3T3 became quiescent in the serum-starved culture conditions. However, a residual amount of centrobin was also detected at the centrosomes of the resting cells, suggesting its role for maintaining integrity of the centrosome, especially of the daughter centriole in the cells.

Toxicity of the Puffer fish, Takifugu xanthopterus (Kkachibok) and Takifugu stictonotus (Kkachilbok) from Coastal Area of Korea (한국 연안산 까치복(Takifugu xanthopterus)과 까칠복(Takifugu stictonotus)의 독성)

  • Kim, Ji-Hoe;Mok, Jong-Soo;Son, Kwang-Tae;Kim, Ju-Gyeong;Jo, Mi-Ra;Kim, Poong-Ho;Lee, Tae-Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.40 no.5
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    • pp.276-281
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    • 2007
  • The toxicity of two species of puffer fish, Takifugu xanthopterus and T. stictonotus, collected from coastal regions of Korea, was determined using a mouse bioassay. The highest toxin scores in the muscle, skin, fins, and testis in both species were below 50 mouse units (MU) per gram, and for each organ of both species the proportion of toxic specimens containing ${\geq}10MU/g$ was less than about 10%. In T. xanthopterus, the highest toxin levels in the liver, gallbladder, and ovary exceeded 1,000 MU/g (1,275-1,910), while less than 200 MU/g (12-136) was detected in the same organs of T. stictonotus. Therefore, the toxicities of muscle, skin, and testis in both species of puffer fish were within acceptable levels for human consumption.

Studies on Utilization and Lipid Composition of Nonedible-Tissues from Fugu xanthopterus (복어의 비가식부의 이용과 지질 조성에 관한 연구)

  • 이민경
    • The Korean Journal of Food And Nutrition
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    • v.10 no.2
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    • pp.213-218
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    • 1997
  • Total lipid were extracted from the eggs, livers, intestines, muscles, and testis of Fugu xanthopterus. The fatty acid composition of total lipid was analyzed by gas liquid chromatography. Proximate percentage of total lipid from the samples was shown to be ; 81.72% in the livers, 14.53% in the intestines, 12.63% in the eggs, 0.85% in the muscles, and 1.74% in the testis. Toxins(tetrodotoxins) were completely removed from the liver of Fugu xanthopterus with 1% acetic acid/methanol solution before total lipid was extracted with CHCI3:methanol solution(2:1, v/v). The toxicity left in the total lipid was checked using mice. The content of DHA in each tissues of Fugu xanthopterus was the most abundant in muscle(32.4%), followed by liver(23.0%), intestine(26.7%) and testis(15.4%). The level of total polyenoic acids comprising DHA was more abundant in muscle(57.4%) than in the liver(50.9%).

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Subacute Toxicity Test of Guh Sung Y.L.S.-95 (Guh Sung Y.L.S.-95의 아급성 독성시험)

  • 김판기;왕성호;김대용
    • Journal of Food Hygiene and Safety
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    • v.12 no.3
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    • pp.234-239
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    • 1997
  • Guh Sung Y.L.S.-95 is one of the polyacidic solution of which main component is acetic acid. We investigated the subchronic toxicity of the Guh Sung Y.L.S.-95 using SPF ICR mouse for 4 weeks. The Guh Sung Y.L.S.-95 was administered by gastric intubation, 1.0, 2.5, 5.0 g/kg body weight. The results are as follows: 1. There are no adverse effects on the clinical obserbation and body weight changes. Also, there are some significant changes in organ weight, but it was meaningless because of the absence of dose-response relationships. 2. In the hematological patterns of administered mouse, there are no significant changes between the treated groups. Also, there are no serological enzymatic changes in the treated mouse. In the 1.0 g/kg treated group, ASP activity was increased significnatly compared with control group. But, this level of activity was fall under the normal physiological range of control mouse. 3. Histopathological findings of the brain, liver, heart, spleen, kidneys, stomach, lung, testis, ovary, uterus and thymus were not observed in the treated mouse. From the above results, the Guh Sung Y.L.S.-95 has no toxicity upto the 5.0 g/kg/day of oral dose for 4 weeks.

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Effects of Postnatal Exposure to Octylphenol on the Transcriptions of Steroidogenic Enzymes in Mouse Testis

  • Kim, Suel-Kee;Lee, Ho-Joon;An, Su-Yeon;Lee, Chang Joo;Yoon, Yong-Dal
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.550-558
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    • 2004
  • The effects of postnatal exposure to octylphenol(OP) on the expressions of the steroidogenic enzymes and testosterone production were evaluated. Postnatal male mice (15-day-old) were injected with 2 or 20mg $kg^{-l}$ body weight (BW) of OP for 5 days and sacrificed on postnatal day 21. Testosterone concentration was measured by radioimmunoassay and the expressions of the testicular genes were determined by RT-PCR analyses. Significant reductions in the mean body and testis weight were observed in the OP treated animals. No marked alteration in the histological structure of the testis were observed, however, slight reduction in the seminiferous tubule diameter and the number of Leydig cells and several pyknotic cells could be identified in the 20 mg $kg^{-l}$ BW of the OP treated animals. Serum testosterone concentration was dramatically reduced and the mRNA expressions of the steroidogenic acute regulatory protein (StAR), cholesterol side-chain cleavage enzyme (P450scc) and $17\beta$-hydroxylase/Cl7-20 lyase $(P450_{17\alpha})$ were decreased. No significant changes of the gene expressions of the steroidogenic factor-l (SF-I) and estrogen and androgen receptor after the OP treatment showed that the decreased expressions of the steroidogenic enzymes in the present study did not correlate with these genes. Altogether, the present study demonstrates that postnatal treatment of OP inhibits steroidogenesis by decreasing the transcriptional expressions of the StAR and steroidogenic enzymes. The alteration in steroidogenesis may adversely affect the normal development of the testis and sper- matogenesis.

Hatching of mouse balstocysts on somatic cell culture

  • Nah, Hee-Young;Gye, Myung-Chan
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.43-44
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    • 1998
  • Effect of somatic cell coculture on hatching of mouse blastocyst was examined. Mid-blastocysts were cocultured with granulosa cell primary culture or Sertoli cell line ($TM_{4}$) derived from mouse testis for 48 hr. Blastocysts cultured in medium (10% FBS) started to hatch more faster than cocultured embryos during 12 hr of coculture. After then blastocysts cocultured with somatic cell hatched faster than control. Degeneration of embryos was also greately reduced by coculture. This result suggested the potentiation of hatching as well as embryonic viability by coculture with somatic cell and Sertoli cell line can be used for embryo coculture.

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EID-1 Interacts with Orphan Nuclear Receptor SF-1 and Represses Its Transactivation

  • Park, Yun-Yong;Park, Ki Cheol;Shong, Minho;Lee, Soon-Jung;Lee, Young-Ho;Choi, Hueng-Sik
    • Molecules and Cells
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    • v.24 no.3
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    • pp.372-377
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    • 2007
  • The orphan nuclear receptor, SF-1, plays a pivotal role in the development and differentiation of the endocrine and reproductive systems, and also regulates the transcription of a host of genes, including those encoding several steroidogenic enzymes and gonadotropins. We found that a previously unidentified repressor, EID-1, is an SF-1-interacting protein that inhibits the transactivation of SF-1. A transient transfection assay revealed that EID-1 inhibits SF-1, but not LRH-1, $ERR{\gamma}$, or mCAR. Using the yeast two hybrid and GST pull-down assays, we determined that EID-1 interacted strongly with SF-1. In addition, it colocalized with SF-1 in mammalian cells and interacted specifically with the AF-2 domain of SF-1, competing with SRC-1 to inhibit SF-1 transactivation. EID-1 is expressed in the mouse testis, and its expression decreases during testis development. The results of the present study suggest that EID-1 can act as a repressor, regulating the function of SF-1.