• Proceedings of the Zoological Society Korea Conference
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• 2001.10a
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• pp.177.3-177
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• 2001

No Abstract, See Full Text

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.02a
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• pp.18-18
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• 2003

No Abstract, See Full Text

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.47-55
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• 2003

Objectives: Bok, Bcl-2-related ovarian killer, is a proapoptotic Bcl-2 family protein identified in the ovary based on its dimerization with the antiapoptotic protein Mcl-1. The present study examined the hormonal regulation and localization of Bok messenger RNA levels in the mouse ovary during the follicle development. Methods: The animals were implanted subcutaneously with Silastic brand capsules containing the synthetic estrogen, DES at $21{\sim}23$ days of age. Ovaries were collected $1{\sim}3$ days after implantation for RNA analysis and in situ hybridization. Some mice were removed capsule for $1{\sim}2$ days to induce ovarian follicle apoptosis. Ovaries were also collected from 26 day-old immature mice at various times after treatment with 10 IU PMSG. Some mice received a single intraperitoneal injection of 10 IU hCG to induce ovulation, and ovaries were obtained at different time intervals for Northern blot and in situ hybridization analysis, respectively. Results: Treatment of immature mice with diethylstilbestrol (DES) for $24{\sim}48$ h increased ovarian Bok mRNA levels. Bok mRNA was remained the same levels in mice removed DES for $24{\sim}48$ h to induce apoptosis. High signals of Bok mRNA after DES treatment were detected in granulosa cells of early antral follicles. Treatment of immature mice with PMSG for 12 h increased markedly ovarian Bok mRNA expression which was detected mainly in preantral and atretic follicles. Interestingly, low levels of Bok mRNA were also expressed in granulosa cells of preovulatory follicles. Treatment of PMSGprimed mice with hCG stimulated strongly ovarian Bok mRNA expression at $6{\sim}9$ h. At that time, Bok mRNA was expressed in granulosa cells of atretic and small growing follicles. Conclusion: These results demonstrate that Bok is one of proapoptotic Bcl-2 members expressed in early growing and atretic follicles during the ovarian follicular development. Gonadotropins induce a transient increase of Bok gene expression in granulosa cells of preantral and preovulatory follicles indicating some role in the ovulatory process.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.4
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• pp.253-263
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• 2006

Objective: Identification of the regulatory mechanism for arrest and initiation of primordial follicular growth is crucial for female fertility. Previously, we found 15 expressed sequence tags (ESTs) that were specifically abundant in the day-S-subtracted cDNA library and that the B357 clone was novel. The present study was conducted to obtain the whole sequence of the novel gene including B357 and to characterize its mRNA and protein expression in mouse ovary and testis. Methods: The extended sequence of the 2,965-bp cDNA fragment for the clone B357 was named ${\underline{5}}-{\underline{d}}ay-{\underline{o}}vary-{\underline{s}}pecific\;gene-{\underline{1}}$ (5DOS1) and submitted to GenBank (accession number ${\underline{AY751521}}$). Expression of 5DOS1 was characterized in both female and male gonads at various developmental stages by Northern blotting, real-time RT-PCR, in situ hybridization, Western blotting, and immunohistochemistry. Results: The 5DOS1 transcript was highly expressed in the adult testis, brain, and muscle as compared to the other tissues. In the ovary, the 5DOS1 transcript was detected in all oocytes from primordial to antral follicles, and highly expressed at day 5 after birth and decreased thereafter. In contrast, expression of 5DOS1 showed a gradual increase during testicular development and its expression was limited to various stages of male germ cells except spermatogonia. Conclusions: This is the first report on the expression and characterization of the 5DOS1 gene in the mouse gonads. Further functional analysis of the 5DOS1 protein will be required to predict its role in gametogenesis.

• Development and Reproduction
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• v.4 no.1
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• pp.13-17
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• 2000

Follicle stimulating hormone (FSH) stimulates follicle growth, and inhibits the follicle atresia in the immature rodent ovaries. The present study was carried out to know the histological changes of ovarian follicles after FSH treatment in the prepubertal mice. Ten i.u. of recombinant FSH was i.p. injected on 3 weeks old mice. After the treatment, at 1, 2 and 3 days, left ovaries were collected for the histological study. The atretic ratio of preantral follicles increased with time after FSH treatment. However, in the case of antral follicles, there was no significant change in the ratio. The degenerating follicles contained apoptotic granulosa cells, macrophage, and polymorphonuclear leukocytes in the follicular cavity. The present results suggest that follicular degeneration caused by FSH hyperstimulation could be mediated by apoptosis as well as the acute inflammation.

• Development and Reproduction
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• v.14 no.4
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• pp.287-295
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• 2010

It was recently reported that nesfatin-1/NUCB2, which is secreted from the brain, controls appetite and energy metabolism. The purpose of this research was to confirm whether or not the protein and its binding site should have been expressed in the mouse reproductive organs and to know the possible effects of nesfatin-1 on the reproductive function. Using the ICR female mouse ovary and uterus, the expression of NUCB2 mRNA was confirmed with the conventional PCR and the relative amount of NUCB2 mRNA in the tissues was analyzed with real-time PCR. Immunohistochemical staining was performed using the nesfatin-1 antibody to investigate the nesfatin-1 protein expression and the biotin conjugated nesfatin-1 to confirm the binding site for nesfatin-1 in the ovary. Furthermore, in order to examine if the expression of NUCB2 mRNA in the ovary and uterus is affected by gonadotropin, its mRNA expression was analyzed after PMSG administration into mice. As a result, the expression level of NUCB2 mRNA in the ovary and the uterus was as much as the expression level in hypothalamus. As a result of the immunohistochemical staining, nesfatin-1 proteins were localized at the theca cells, the interstitial cells, and some of the luteal cells. However, the granulosa cells in the follicles did not stain. Interestingly, the oocytes in the some follicles were stained with nesfatin-1. On the other hand, nesfatin-1 protein binding sites were displayed at the theca cells and the interstitial cells near the tunica albuginea. After PMSG administration the expression level of NUCB2 mRNA was increased in the ovary and the uterus. These results demonstrate that for the first time the nesfatin-1 and its binding site were expressed in the ovary and NUCB2 mRNA expression was controlled by gonadotropin, suggesting an important role in the reproductive organs as a local regulator. Therefore, further study is needed to elucidate the functions of nesfatin-1 on the reproductive organs.

• Development and Reproduction
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• v.7 no.2
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• pp.119-126
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• 2003

In the previous study, we obtained list of differentially expressed genes between postnatal day 1 and day 5 mouse ovaries using suppression subtractive hybridization(SSH) and found that MT Oansposon-like element, clone MTi7(MTi7) was one of the highly expressed genes in the day 5 mouse ovary(Park et at., 2002). Results of in situ hybridization and RNA interference revealed that the expression of MTi7 is oocyte-specific in the ovary and may be involved in the regulation of oocyte maturation(Park et at., 2003). At present, MTi7 sequence has been known only in the mouse. Therefore, the present study was accomplished 1) to identify MTi7 sequence in the other mammalian species, such as bovine, porcine, rat, and human, and 2) to evaluate the flanking sequence of the mouse MTi7 since it has transposon characteristics. Using ovarian cDNAs derived from low different species, we cloned and identified new MTi7 sequence showing a high degree of sequence homology with the mouse MTi7(87∼98%). By using inverse PCR, we found that the mouse MTi7 may intercalated the beta-carotene 15, 15'-monooxygenase(Bcdo) gene and/or serine protease inhibitor, Kunitz Dpe I(Spint 1) gene. By finding the MTi7 sequences in the other mammalian species and the flanking gene of the MTi7 in mouse, it is expected to reveal the role(s) of MTi7 in the oogenesis as well as folliculogenesis in the near future.

• 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.

• Clinical and Experimental Reproductive Medicine
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• v.33 no.1
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• pp.35-35
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• 2006

Objective: The purposes of the present study were to investigate the effect of ${\gamma}$-radiation on the expression of inhibin-${\alpha}$ proteins and genes for inhibin ${\alpha}$, ${\beta}A$, and ${\beta}B$ in the ovary. Methods: Immature mice were whole-body ${\gamma}$-irradiated with 25% of a lethal dose. At time 0, 3, 6, 12, and 24 hours after the irradiation, the ovaries were collected and used for immunohistochemistry for inhibin-${\alpha}$, and RT_PCR for inhibin-${\alpha}$, ${\beta}A$, and ${\beta}B$. Results: The expression of the immunoreactive inhibins-${\alpha}$ was maintained at 12 hours post-irradiation and reduced thereafter. The expression of inhibin-${\alpha}$ mRNA was significantly increased with the time after the irradiation. However there were no significant changes in the expression of ${\beta}A$ and ${\beta}B$ mRNAs. Conclusion: It might be thought that inhibin acts as one of the regulatory factors in the ${\gamma}$-radiation-induced follicular atresia in mice

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.75-75
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• 2003

Transition of the resting primordial follicle to the growing primary follicle is a critical process for female reproduction, but its mechanism is poorly understood. The present study was conducted to investigate gene expression profile at the primordial-primary follicle transition process. We isolated total RNA of female mouse ovary at day1 (contains only primordial follicles) and day5 (contains primordial and primary follicles) and synthesized cDNA using annealing control primers (ACP; Seegene, Inc., Seoul, Korea). ACP provides annealing specificity and sensitivity to the template and allows to identify only authentic differentially expressed genes (DEGs). We used total 80 ACPs for PCR, observed PCR products on 2% agarose gel, cloned 42 DEGs using TOPO TA cloning vector, sequenced, and analyzed by BLAST search. Sequences of 34 clones significantly matched database entries while 4 clones were novel and 4 clones were EST. Two of 34 genes were specifically expressed only in day 5 ovaries (Sui1-rs1, Apg3p/Aut1p-like), and rest of 32 genes were expressed in both stages but were differential in amount. Differential expression was confirmed using semiquantitative RT-PCR, and there was no false positive. Anx11 and Pepp2-pending were highly expressed genes in day1-, while BPOZ, Ches1, Kcmf1, NHE3, Nid2, Ninj1, SENP3 and Survivin were highly expressed genes in day5-ovary. List of genes would provide insight for further study of mechanism regulating primordial-primary follicle transition.