• Clinical and Experimental Reproductive Medicine
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• v.15 no.2
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• pp.135-147
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• 1988

To compare the stimulation effect of the ratio in follicle stimulating hormone and luteinizing hormone in induction of multiple follicular growth, the serum $E_2$ level, the diameter of follicle, number of aspirated follicles and cleavage rate of in vitro fertilized preovulatory oocytes as well as the pregnancy rate were evaluated. Forty one patients with irreparable tubal disease were stimulated by hMG(n=24) or FSH/hMG(n=17) for the purpose of in vitro fertilization and embryo transfer. The following results were obtained. 1. Serum estradiol($E_2$) levels on the day of hCG administration were $921.0{\pm}353.3\;pg/ml$ in hMG group and $1272.9{\pm}1060.6\;pg/ml$ in FSH/hMG group. The serum $E_2$ value of hMG group was significantly lower than that of FSH/hMG group. 2. The diameter of leading follicle by ultrasonogram on the day of hCG administration were $16.2{\pm}2.0\;mm$ in hMG group and $16.2{\pm}2.6\;mm$ in FSH/hMG group. No significant difference of follicle diameter between two groups was demonstrated. 3. The number of follicles with diameter above 10 mm by sonogram on the day of hCG injection were $3.91{\pm}2.32$ in hMG group and $6.52{\pm}3.86$ in FSH/hMG group. There was significant difference of number of follicles between two groups, (p< 0.01). 4. The number of oocytes found per patient at aspiration were $2.59{\pm}1.00$ in hMG group and 3. $76{\pm}2.31$ in FSH/hMG group. There was significant difference of number of aspirated oocytes between two groups. (p< 0.05). 5. The detection rate of preovulatory oocyte at aspiration were 68.4%(39/57) in hMG group (n=22) and 77.6%(38/49) in FSH/hMG group (n=13). 6. The cleavage rate of preovulatory oocyte at 44 hours after insemination were 74.4%(29/39) in hMG group(n=22) and 81.6%(31/38) in FSH/hMG group (n=13). When only hMG was used, one pregnancy was established in 15 patients to whom 29 zygotes were transferred. And a full term normal female baby was delivered by elective cesarean section. In the FSH/hMG group, five pregnancies out of 9 transferred patients were confirmed by serum ${\beta}-hCG$. Two pregnancies were spontaneously aborted before the 6th week of pregnancy. One patient aborted her baby at the 18th week of pregnancy because of incompetent internal os of the cervix. Two patients delivered two full term babies by elective cesarean section. From the above findings, paralell with the increase in the ratio of exogenous follicle stimulating hormone to luteinizing hormone, an increase in oocyte recovery was observed as well as an improvements in pregnancy rate. It was concluded that FSH enrichment early in the follicular phase had a beneficial effect in the controlled ovarian hyperstimulation.

• The Journal of Korean Obstetrics and Gynecology
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• v.21 no.3
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• pp.60-74
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• 2008

Purpose: This study was designed to investigate the effects of Gwibi-Tang(GBT) on the polycystic ovary(PCO) induced by estradiol valerate(EV) in rats. Methods: PCO was induced by single intramuscular injection with EV(4mg) in female rats. Normal group(n=8) were injected with sesame oil and orally administrated distilled water for eight weeks. PCO control group(n=8) were injected with EV and orally administrated distilled water for eight weeks. GBT treated group(n=8) were injected with EV and orally administrated GBT for eight weeks. Then we measured weight of body, ovaries, adrenal glands, and uterus of rats. The histopathology changes of ovaries were also evaluated. The expression of nerve growth factor(NGF) was analyzed in the central nervous system, adrenal glands and ovaries by immunohistochemistry. And also CRF expression in median eminance of Rats were analyzed. Results: 1. The weight(g) of rats in GBT treated group($275{\pm}14$) was significantly increased(p<0.01) compared with control group($253{\pm}8$), 2. The weight(mg) of ovaries in GBT treated group($75.8{\pm}16.7$) was significantly increased(p<0.001) compared with control group($37.4{\pm}6.7$). 3. The number of mature follicles in GBT treated group($3.6{\pm}1.2$) was significantly increased(p<0.01) compared with control group($1.5{\pm}1.5$. 4. The number of atretic follicles in GBT treated group($8.0{\pm}3.1$) was significantly decreased(p<0.01) compared with control group($18.6{\pm}6.0$). 5. The number of cystic follicles in GBT treated group($0.5{\pm}0.5$) was significantly increased(p<0.01) compared with control group($2.3{\pm}1.3$). 6. The number of corpora lutea in GBT treated group($6.1{\pm}3.9$) was significantly increased(p<0.01) compared with control group($1.6{\pm}2.3$). 7. The expression of NGF-immunoreactive cells in the ovarian granulosa cells in GBT treated group was lesser observed than control group. Conclusion: From the above results, we concluded that Gwibi-Tang has inhibitory effect on the development of EV-induced polycystic ovary. And it's effect may be related with decreased NGF activities in the ovary.

• Animal cells and systems
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• v.5 no.1
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• pp.45-50
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• 2001

Rana ovarian follicles consist of oocyte, vitelline envelope, granulosa cells, and theca/epithelial layer. Using scanning electron microscopy, the surface structure of each follicular component was investigated. Changes in oocyte surface during oocyte maturation were also examined. Theca/epithelial layer was almost transparent and some blood vessels and granulosa cells were observed underneath in intact follicle. The number of granulosa cells was estimated to be 6700-7200 per oocyte. The granulosa cells partially overlapped each other and their microvilli penetrated the vitelline membrane via holes present in the vitelline envelope and seemed to be linked to oocyte microvilli. After removal of the vitelline envelope by microforcep, oocyte microvilli were observed on the surface of the devitellined oocyte. The oocyte microvilli formed partial clusters on the surface of white spot area which appears iust before germinal vesicle breakdown (GVBD), whereas they were evenly distributed in other areas. The microvilli became shorter and less dense with oocyte maturation. The lengths of oocyte microvilli in the immature and mature oocyte were 1.5 $\mu$m and 0.6 $\mu$m, respectively. The present study suggests a fundamental structural change occurring on the oocyte surface during maturation.

• Korean Journal of Animal Reproduction
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• v.14 no.1
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• pp.31-35
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• 1990

The viability and developmental potential of rat oocyte was assessed by the use of eosin B. Dye was prepared as a stock solution to contain 1.0 mg eosin B per ml of Brinster's Medium for Ovum Culture plus glucose (BMOC) -2 medium. Three dye concentrations were selected for the Oocytes from rat ovarian follicles were collected and assessed under various conditions of heat oocyte studies by dilution of the stock solution with BMOC: 0.12mM, 0.60mM and 1.20mM. treatment and preservation duration by dye exc¬lusion assay using eosin B. It is suggested that live oocyte excludes eosin B whereas dead oocyte does not.

• Korean Journal of Animal Reproduction
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• v.21 no.2
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• pp.117-122
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• 1997

The aim of this study was to investigate the effect of the follicular culture from which the oocytes originate on their subsequent in vitro maturation ability. Ovarian follicles were isolated and cultured according to size(1~2mm, 2~6mm and 6~8mm) for 42~44 h. The rates of germinal vesicle breakdown(GVBD) in each groups were 87%(65/75), 82%(80/97) and 89%(47/53), but the oocytes maturation were su, pp.essed at anaphase-I stage. In spite of the adding porcine follicular fluid and/or hormones in maturation medium, maturation ability of oocytes from follicle cultured for 21~22 h were inhibited. When oocytes from follicle cultured for 4 h at various temperature were incubated for 38~40 h, the rates of oocytes maturation from follicle cultured at 2$0^{\circ}C$(51%, 26/51) and 39$^{\circ}C$(54%, 26/48) were significant higher(P<0.05) than group cultured at 4$^{\circ}C$(33%, 19/58). On the other hand, the GVBD were stared 2 h after culture of follicle of oocytes. To summairze, oocytes maturation by follicular culture were inhibited at anaphase-I stage in porcine. When the follicle cultured for 4 h, maturation were completed to metaphase-II stage. However, rates of GVBD in oocytes from follicular culture were higher than oocytes cultured in medium.

• Journal of Pharmaceutical Investigation
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• v.23 no.3
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• pp.31-40
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• 1993

It is essential to clone the peptide transporter in order to obtain better understanding of its molecular structure, regulation, and substrate specificity. Characteristics of an endogenous peptide transporter in oocytes were studied along with expression of an exogenous proton/peptide cotransporter from rabbit intestine. And further efforts toward cloning the transporter were performed. The presence of an endogenous peptide transporter was detected in Xenopus laevis oocytes by measuring the uptake of $0.25\;{\mu}M\;(10\;{\mu}Ci/ml)\;[^3H]-glycylsarcosine$ (Gly-Sar) at pH 5.5 with or without inhibitors. Uptake of Gly-Sar in oocytes was significantly inhibited by 25 mM Ala-Ala, Gly-Gly, and Gly-Sar (p<0.05), but not by 2.5 mM of Glu-Glu, Ala-Ala, Gly-Gly, Gly-Sar and 25 mM glycine and sarcosine. This result suggests that a selective transporter is involved in the endogenous uptake of dipeptides. Collagenase treatment of oocytes used to strip oocytes from ovarian follicles did not affect the Gly-Sar uptake. Changing pH from 5.5 to 7.5 did not affect the Gly-Sar uptake significantly, suggesting no dependence of the endogenous transporter on a transmembrane proton gradient. An exogenous $H^+/peptide$ cotransporter was expressed after microinjection of polyadenylated messenger ribonucleic acid $[poly\;(A)^+-mRNA]$ obtained from rabbit small intestine. The Gly-Sar uptake in mRNA-injected oocytes was 9 times higher than that in water-injected oocytes. Thus, frog oocytes can be utilized for expression cloning of the genes encoding intestinal $H^+/peptide$ cotransporters. Using the technique size fractionation of mRNA was sucessfully obtained.

• Korean Journal of Veterinary Research
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• v.48 no.3
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• pp.323-326
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• 2008

A two-year-old mixed breed sow was requested to the Veterinary Pathology Laboratory of Cheju National University with a clinical signs of severe abdominal pain and sudden death. Grossly, there was severe hemorrhage in abdominal cavity. Most of internal parenchymas and subcutaneous muscle showed severe pale discoloration. Both ovaries were enlarged with oval to round protruding multilobular masses and dark red in color. And they were firm and contained multiple small cysts in their cut surface. Histopathologically, numerous neoplastic granulosa cells had spherical-to-oval, hyperchromatic nuclei and scant eosinophilic cytoplasms were distributed with follicular pattern in ovarian masses. And the typical Call-Exner bodies, distinctive microcavityies, were observed in the center of small neoplastic follicles. Based on the gross and histopathologic findings, this case was diagnosed as granulosa cell tumor. In our best knowledge, this is believed to be the first report of granulosa cell tumor in a sow in Korea.

• Korean Journal of Veterinary Pathology
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• v.1 no.1
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• pp.72-76
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• 1997

Histological investigation of the number of follicles following gonadotropin treatments for superovulation was carried out in mature Sprague-Dwaley(SD) rats. Routinely serial sections of paraffin-embedded ovaries were stained with hematoxylin-eosin and evaluated with light microscope. During the study unusual cases of microscopic alterations were observed in the medulla of ovaries in two rats. Case one: An ovum and its follicular fluid outflowed in medulla of ovary. The follicular fluid was densly proteinuous. Corona raiata consisted of 2-6 layers thick cells in the periphery of the ovum. While the cortical side of the follicular wall was intact with normal granulosa cell layer the meullary side of it was ruptured. Case two: A large cyst was present in medulla of ovary hilus. The cyst occupied the entire medulla displacing the ovarian archetecture and enclosed by connective tissue and smooth muscle wall.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.47 no.6
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• pp.882-887
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• 2014

We studied oocyte steroidogenesis in blacktip grouper Epinephelus fasciatus ovarian follicles during vitellogenesis. Vitellogenic oocytes with average diameters of 0.45, 0.48 and 0.50 mm were incubated in vitro in the presence of $[^3H]17{\alpha}$-hydroxyprogesterone as a precursor. The steroid metabolites were analyzed using thin layer chromatography (TLC), high performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC/MS). The major metabolites in the vitellogenic oocytes were androstenedione ($A_4$), testosterone (T), estradiol-$17{\beta}$ ($E_2$), and estrone ($E_1$). The metabolites of androgen ($A_4$ and T) were higher in the 0.50-mm oocytes than in the 0.45- and 0.48-mm oocytes, while the estrogen metabolites (E2 and E1) were lower in the 0.50-mm oocytes. These results suggest that 0.50-mm oocytes are fully vitellogenic following initiation of the maturation process.

• 대한생식의학회:학술대회논문집
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• 2006.06a
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• pp.6-11
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• 2006

Our previous study demonstrates a rapid activation of atypical PKC${\zeta}$ by the ovulatory dose of LH/hCG. The present study was therefore designed to identify PKC${\zeta}$ regulated-genes in rat ovarian preovulatory granulosa cells. Preovulatory granulosa cells cultured in the presence of myristoylated PKC${\zeta}$ pseudosubstrate peptide were subjected to identify differentially expressed genes by using anneling control primer RT-PCR. As a result, among sixteen genes identified, six genes (testin, glypican-4, retrovirus SC1, connective growth factor, aminolevulinic acid synthase1 and serum- inducible kinase) were rapidly stimulated by hCG. Northern blot analysis demonstrated that all these genes were rapidly stimulated by hCG and declined thereafter. In situ hybridization analysis revealed the expression of these genes in granulosa cells of preovulatory follicles. The present study demonstrates time- and cell-specific expression of PKC${\zeta}$-regulated genes, and may imply that these genes play a specific role(s) during LH-induced ovulation.