• 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.

• Development and Reproduction
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• v.17 no.3
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• pp.269-274
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• 2013

This study was performed to evaluate the effects of fibroblast co-culture on in vitro maturation (IVM) of prepubertal mouse preantral follicles. The intact preantral follicles were obtained from the ovaries of 12-14 day old mice and these were cultured individually in ${\alpha}$-minimal essential medium (${\alpha}$-MEM) supplemented with 5% fetal bovine serum (FBS), $100mIU/m{\ell}$ recombinant follicle stimulating hormone (rFSH), 1% insulin-transferrin-selenium, $100{\mu}g/ml$ penicillin and $50{\mu}g/m{\ell}$ streptomycin as base medium for 12 days. A total of 200 follicles were cultured in base medium co-cultured with mouse embryonic fibroblast (MEF) (MEF group) (n=100) or only base medium as control group (n=100). Survival rate of follicles on day 12 of culture were significantly higher in the MEF group of 90.0%, compared with 77.0% of the control group (p=0.021). Follicle diameters on day 6 and 8 of the culture period were significantly larger in the MEF group than those in the control group (p=0.021, p=0.007, respectively). Estradiol levels in culture media on day 4, 6, 8, 10 and 12 of the culture period were significantly higher in the MEF group (p=0.043, p=0.021, p=0.006, p<0.001 and p=0.008, retrospectively). Our data suggest that MEF cell co-culture on IVM of mouse preantral follicle increases survival rate and promotes follicular growth and steroid production.

• Korean Journal of Animal Reproduction
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• v.5 no.1
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• pp.43-63
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• 1981

This study aimed to determine the effects of adrenalectomy on the sexual gland, the thyroid gland and the serum components. A total of 192 Wister strain albino rats were evenly divided into 4 sexually equal groups for the comparisons. Each groups was divided into a control and a treatment group by sexuality. Each tissue of the sexual gland and the thyroid gland was microscopically examined, and at the same time body weight and serum components were also examined on the 1st, 7th, 14th, 28th, 42nd, 56th, 70th and 84th day respectively following adrenalectomy. The results obtained were as follows: 1. The body weight was sufficiently retarded in decreasingly after the adrenalectomy as compared to that of control. 2. The histological change of testis started atrophy of spermatogonia and degeneration of spermatocyte. There was a, pp.ared no cell division activity. Spermatozoa in the seminiferous tuble was not noticed but degeneration of interstitial cell was started and spermatozoa in the epididymal duct disa, pp.ared. 3. Degeneration of oocyte and follicular cell was noticed as the histological change of ovary. There was not a, pp.arance of primary follicle and corpus Iuteum but interstitial cell was proliferated. 4. For the effect of adrenalectomy on the histological change of the thyroid gland, the number of small follicle decreased and that of large follicle increased as the time passed following the adrenalectomy. And the follicular epithelial cell became squamous as typical condition of functional decreased. 5. It was shown that in the total contents of serum protein no difference with control occurred with in the 70th day for male and female adrenalectomized, respectively. But the differences in protein contents were significanly decreased on the 70th day. 6. No difference occurred in the total serum lipids on the 7th day, but they decreased significantly on the 42nd day, and (P<0.01) on the 56th, and 70th day. A tendency to decrease was noted as time elapsed following adrenalectomy. 7. Increase and decrease were intercrossed in the serum cholesterol contents between the control and th treatment groups on the 28th day. But the difference significantly decreased on the 42nd, 56th, and 70th day after adrelalectomized. The contents showed tendency to decrease as time elapsed following adrenalectomy. 8. The blood glucose contents rapidly decreased with time. The difference were significant on the 28th and 42nd day, and highly significant on the 56th and 70th day for male adrenalectomized. 9. In case of solium, there was no noticeable sexual distinction. There was slight in creasing or decreasing for the control group. But the treatment group tended continuously to decrease for male and female. 10. It was shown that potassium contents tended to increase on the 28th day for male and female, but the differences were small. 11. As for chlorine, it tended to decrease rapidly on 7th day for male and female adrenalectomized, and the tendency continued.

• The Korean Journal of Zoology
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• v.39 no.3
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• pp.257-265
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• 1996

Previously, we demonstrated that estradiol (E$_2$) was produced by medium sized follicles of Rana nigromaculata and Rana dybowskii in vitro. Present experiments were carried out to determine when the growing follicles have obtained the ability to produce E$_2$. Follicles in different growth stages were isolated and cultured for 6 hours in the presence or absence of frog pituitary homogenates (FPH, 0.1 pituitary/2m1) or various steroid precursors (200 ng/2m1). levels of progesterone (P$_4$), 17 -hydroxyprogesterone (17$\alpha$-OHP), androstenedione (AD), testosterone Cr) or E$_2$ in the medium were measured by RIA. The smallest follicles failed to produce steroids, whereas the smaller follicles produced considerable amounts of steroids (211 pg/follicle), and the medium sized follicles produced a large amounts of steroids (1653 pg/follicle) in response to FPH. Addition of pregnenolone (P5) resulted in a marked increase in P$_4$ but not in other steroids by the smallest follicles whereas the treatment resulted in a marked increase in P$_4$, 17$\alpha$-OHP, AD, T and E$_2$by the smaller and medium follicles. When the amounts of steroids are calculated on the basis of unit surface area (pg/mm$_2$), the ability of the smallest follicles to produce P$_4$ from P5 was similar to those of smaller and medium sized follicles. However, the smallest follicles failed to metabolize P$_4$ to other steroids whereas the smaller and medium follicles did. Taken together, the data suggest that the smallest follicles do not response to FPH in terms of steroid production but they have capacity to convert P5 to P$_4$ and that the smaller follicles have potential to produce E$_2$ although much less efficient than medium sized follicles.

• Fisheries and Aquatic Sciences
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• v.1 no.1
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• pp.82-93
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• 1998

Monthly changes of the gonad follicle index (GFI), reproductive cycle, egg-diameter composition, first sexual maturity of the Pacific oyster, Crassostrea gigas, were studied based on the samples which have been collected from the intertidal zone of Poryong west coast of Korea, from January to December, 1996. C. gigas, is dioecious, while a few individuals are alternatively hermaphroditic. Monthly variation of gonad follicle index (GFI) used for determination of spawning period, coincided with the reproductive cycle. GFI increased from April when seawater temperatures gradually increased and reached the maximum in May. And then, GFI sharply decreased from June to September due to spawning. Reproductive cycle of this species can be divided into five successive stages: in females, early active stage (March to April), late active stage (April to May), ripe stage (May to August), partially spawned stage (June to September) and spent/inactive stage (September to February); in males, early active stage (February to March), late active stage (April to May), ripe stage (May to September), partially spawned stage (June to September) and spent/ inactive stage (September to February). The diameter of fully mature eggs are approximately 50um. Spawning occurred from June to September, and two spawning peaks were observed in June and August when the seawater temperature was above $20^{\circ}C$. Percentages of the first sexual maturity of males of 20.1-25.0 mm in shell height were over $50\%$, while those of females of 25.1-30.0 mm in shell height were over $50\%$. All the males of > 30.1 mm and all the females of ^gt; 35.1 mm completed their first sexual maturity. The results suggest that C. gigas has a protandry phenomenon. Sex ratios of 919 oysters observed were 453 females $(49.29\%)$, 429 males $(46.68\%)$, 16 hermaphrodites $(1.74\%)$, and 21 indeterminate individuals $(2.29\%)$. In age class I, sex ratio of males were $64.00\%$, thus, a higher percentage than that of females. It was noted that $64.00\%$ of the young males (age class I) were more functional than females in age class I, but 2-3 year-old oysters showed higher percentage of females. Percentages of hemaphrodites in 2-3 year classes were relatively higher than those in other year classes. Histological pattern of hermaphrodites can be divided into two types: Type I (hermaphrodite having a number of newly formed developing oocytes on the oogenic tissues within a degenerating spermatogenic follicle after discharge of numerous spermatozoa) and Type II (hermaphrodite having two separate follicles in the same gonad).

• Clinical and Experimental Reproductive Medicine
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• v.42 no.2
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• pp.62-66
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• 2015

Objective: To evaluate the effect of a gonadotropin-releasing hormone (GnRH) antagonist protocol using corifollitropin alfa in women undergoing assisted reproduction. Methods: Six hundred and eighty-six in vitro fertilization-embryo transfer (IVF)/ intracytoplasmic sperm injection (ICSI) cycles were analyzed. In 113 cycles, folliculogenesis was induced with corifollitropin alfa and recombinant follicle stimulating hormone (rFSH), and premature luteinizing hormone (LH) surges were prevented with a GnRH antagonist. In the control group (573 cycles), premature LH surges were prevented with GnRH agonist injection from the midluteal phase of the preceding cycle, and ovarian stimulation was started with rFSH. The treatment duration, quality of oocytes and embryos, number of embryo transfer (ET) cancelled cycles, risk of ovarian hyperstimulation syndrome (OHSS), and the chemical pregnancy rate were evaluated in the two ovarian stimulation protocols. Results: There were no significant differences in age and infertility factors between treatment groups. The treatment duration was shorter in the corifollitropin alfa group than in the control group. Although not statistically significant, the mean numbers of matured (86.8% vs. 85.1%) and fertilized oocytes (84.2% vs. 83.1%), good embryos (62.4% vs. 60.3%), and chemical pregnancy rates (47.2% vs. 46.8%) were slightly higher in the corifollitropin alfa group than in the control group. In contrast, rates of ET cancelled cycles and the OHSS risk were slightly lower in the corifollitropin alfa group (6.2% and 2.7%) than in the control group (8.2% and 3.5%), although these differences were also not statistically significant. Conclusion: Although no significant differences were observed, the use of corifollitropin alfa seems to offer some advantages to patients because of its short treatment duration, safety, lower ET cancellation rate and reduced risk of OHSS.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.98-98
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• 2002

Recruitment of primordial follicles(PMF) is crucial for female fertility. however, factors and mechanisms that regulate this process is poorly understood. The present study was conducted to obtain an inclusive view of the gene expression and to identify novel factors and their pathways of regulating PMF arrest and/or growth initiation. Ovaries from one-day neonatal(consists of oocyte and PMF) and five-day old(consists of PMF and primary follicles, PRIF) mice were collected, either total RNA or mRNA was isolated, and suppression subtractive hybridization(SSH) was used to isolate and clone genes that differentially expressed in day 1 and day 5 ovaries. Confirmation that some of these genes are differentially expressed in PMF and/or in PRIF was accomplished by using laser captured microdissection(LCM), RT-PCR. in situ hybridization(ISH) and/or immunohistochemistry(IHC). In toto, 357 clones were sequenced and analyzed by BLAST and RIKEN program. Sequences of 330 clones significantly matched database entries while 27 clones were novel. Forty-two and 47 different genes were identified as differentially expressed in day 1 and day 5 ovaries, respectively, while 7 genes were expressed in both stages of ovaries. Day 5-subtracted library included several genes known as markers far growing follicles, such as ZP2, MATER, and fetuin. Among the genes with assigned functions, 23.8% was associated with cell cycle/apoptosis regulation, 7.1% with cellular structure, 11.9% with metabolism, 26.2% with signal transduction, and 31.0% with gene/protein expression in day 1; while 10.6%, 17.0%, 23.5%, 25.5%, and 23.4% in day 5, respectively. Genes such as GDF-8, Lats2, Septin2, and Weel were the highly expressed genes in PMF, while HSP84, Laminin2, MATER, MTi7, PTP, and Wrn were highly expressed genes in PRIF. We have successfully discovered list of genes expressed in day 1 and day 5 ovaries and confirmed that some of them are differentially expressed in PMF and/or PRIF. Gene expression profile from the present study would provide insight for the future study on the mechanism(s) involved in primordial-primary follicular transition. This work was Supported by Korean Health 21 RND Project, Ministry of Health and Welfare, Korea (01-PJ10-PG6-01GN13-0002).

• Korean Journal of Medicinal Crop Science
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• v.3 no.1
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• pp.25-29
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• 1995

In order to obtain the basic information on cross breeding of peony, the growth and floral characteristics were investigasted for local collected strains in main cultvation area in Korea. Local peony strains were classified by early emergence lines and late emergence lines according to the emergence date. Flower color of single-petaling flower was varied with red, pink and white, on the other hand, red+red, pink+white, pink+red and pink+pink in double-petaling flower. Number of pistil of the most local peony strains was 3-4/flower, number of stamen was varied from 0 to more than 200 ea/flower, classifing by 4 groups according to the diameter of flower. Most of the cross combinations showed the cross comptibility, and number of follicle was 3-5 ea/flower, average number of seed was 15.6 ea/follicle, seed size was grouped : by small grains with less than 10g, medium grains with 10-15g and large grains with more than 15g of 100 seed weight.

• KSBB Journal
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• v.22 no.2
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• pp.109-113
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• 2007

Suspension culture of recombinant Chinese hamster ovary (CHO) cells producing follicle-stimulating hormone was performed to investigate the effect of glycine betaine on cell growth and FSH production at low culture temperature. At 28$^{\circ}C$, cell growth was suppressed, but cell viability remained high for a longer culture period. When the culture temperature was lowered from 37$^{\circ}C$ to 28$^{\circ}C$, more than 14-fold increase in the maximum FSH titer was achieved. In batch culture at 28$^{\circ}C$, the use of 15 mM glycine betaine (GB) to culture medium resulted in the enhancement of maximum cell density and FSH titer by 11% and 17%, respectively, compared to the culture without GB. In pseudo-perfusion culture at 28$^{\circ}C$ with the exchange of fresh medium containing 15 mM GB, a final FSH of $2,058{\mu}g$ which is approximately 1.4-fold higher as compared to the culture without GB was obtained. This enhanced FSH production with 15 mM GB was not just because of enhanced specific FSH productivity (qFSH), but mainly because of the extended culture longevity. Taken together, this result demonstrates that the application of GB at low culture temperature is feasible to enhance the production of recombinant proteins in rCHO cells.

• Clinical and Experimental Reproductive Medicine
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• v.37 no.3
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• pp.253-259
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• 2010

Objective: To evaluate if there is any correlation between the growth rate of dominant follicles and clinical characteristics or outcome variables in women undergoing controlled ovarian hyperstimulation (COH). Methods: This study was performed in 313 in vitro fertilization (IVF) cycles. Follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E2) were measured on day 3 of menstrual cycle, and serial ultrasonographic measurement of the diameter of growing follicles was performed. The growth rates of dominant follicles calculated by diameter difference divided by days were correlated with clinical characteristics and outcome variables. Results: There was no significant difference in the growth rate of the dominant follicles between gonadotropin releasing hormone (GnRH) agonist and antagonist cycles. No significant correlation was found between the growth rates and evaluated factors such as age, body mass index, LH, FSH, $E_2$, retrieved oocytes and fertilization rate. Conclusion: The Growth rate of dominant follicles seems to show an independent feature of basal characteristics and ovarian response.