• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.35-40
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• 1989

In order to study the follicular atretic mechanism in mammalian ovary, the relationship between pyknotic index (PI) of granulosa cells and the steroid concentrations in the follicular fluid of atretic follicle was investigated. Follicles were isolated from porcine ovary according to their sizes and the reproductive phases. Steroid concentrations were quantified by high-performance liquid chromatography. As PI increased, the concentration of progesterone was significantly increased(p<0.05), whereas testosterone and estradiol showed no significant changes in their concentration. As follicular size was increased, PI of follicular GC in the luteal phase was increased significantly(p<0.05) and the molar ratio of progesterone to testosterone was increased in the follicles of follicular phase. It can be concluded that progesterone accumulated in the follicular fluid as atresia of the follicles was progressed, and that PI of granulosa cells could be used as one of convenient and pratical criteria for the identification of follicular atresia.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.31-37
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• 1997

Ultrasonographic diagnosis of genital disease and early pregnancy diagnosis was performed in Korean native cattle. The size of ovarian follicle in preovulation, luteal stage and follicular cyst was 18.9, 9.2 and 27.6 mm, respectively, and the thickness of follicular wall was 2.3, 1.8 and 2.8 mm, respectively. The size of corpus luteums in formation stage, activity stage, regression stage, cystic corpora lutea and luteal cyst was 6.2, 11.3, 8.6, 26.7 and 25.9 mm, respectively. The thickness of luteal wall in cystic corpora lutea and luteal cyst was 8.4 and 4.9 mm, respectively. The size of embryo or fetus on day 25, 27, 30, 35, 40, 45 and 50 was 0.8, 0.9, 1.3, 1.5, 2.2, 2.8 and 3.8 cm, respectively. The size of amniotic vesicle on day 25, 27 and 30 was 1.2, 2.1 and 3,0 cm, respectively. The diameter of pregnant uterus on day 25 and 27 was 7.0 and 7.8 cm, respectively. It was concluded that the ultrasonographci values determined in this study can be used as references for the treatment of genital disease and early pregnancy diagnosis in Korean native cattle.

• 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.

• The Korean Journal of Zoology
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• v.33 no.2
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• pp.152-157
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• 1990

The present study, using immunohistochemical procedure, was carried out to determine the localization of immunostainable $\beta$-endorphin cells in the mouse ovarian tissues. Mature female mice were perfused with 4% neutral buffered paraformaldehyde under anesthesia and then frozen-sections were immunostained with anti $\beta$-endorphin antiserum according to ABC technique. Immunoreactive $\beta$-endorphin was found in the luteal cells of corpus lutea, but not in the thecal cells. More strong immunostaining signak were observed in large corpus luteum, in particular, the regressing luteal cells. Primary and secondary follicles did not show any immunoreactivity of $\beta$-endorphin, but granulosa cells lining the antral cavity of large antral follicles contained immunoreactive $\beta$-endorphin.

• Clinical and Experimental Reproductive Medicine
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• v.17 no.1
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• pp.29-44
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• 1990

In 105 patients with the past history of poor response to the previous controlled ovarian hyperstimulation(COH) due to poor follicular growth or premature LH surge, the effectiveness of pituitary suppression with gonadotropin-releasing hormone agonist(GnRH agonist) in IVF/GIFT program was evaluated in 112 cycles of COH using a combination regimen of Leuprolide acetate (Lupron TAP Pharmaceuticals, USA) and FSH/hMG or pure FSH from May to December, 1989 at SNUH. Starting on day 21 of the menstrual cycle(MCD #21, Day 1), Lupron (1.0mg/day, subcutaneous) was administered once a day till next MCD #3(suppression phase). After the confirmation of pituitary suppression, ovarian follicular growth was stimulated with FSH/hMG or pure FSH from MCD #3(Day + 1), and Lupron was continued with hMG or FSH until hCG administration (D 0) (stimulation phase). After suppression phase, serum E2 level decreased from 183.7${\pm}$95.1(Day 1) to 17.4${\pm}$12.3pg/ml (Day +1), and serum progesterone level from 19.17${\pm}$8.67 to 0.12${\pm}$0.05ng/ml. But there was no decresas in serum LH and FSH levels; LH from 12.74${\pm}$6.21 to 15.49${\pm}$4.93mIU/ml,FSH from 7.60${\pm}$3.84 to 8.58${\pm}$3.15 rnlU/ml. There was no occurrence of premature LH surge during COH. Eleven cycles(9.8%) were cancelled due to poor follicular growth during stimulation phase, and 3 cycles (3.0%) failed in the transvaginal oocytes fretrieval. Serum E2 level was 1366.8${\pm}$642.4 on D 0 and 1492.3${\pm}$906.9pg/ml on D+1. 7.00${\pm}$3.32 follicles(FD${\geq}$12mm) were observed on D 0, and 6.11${\pm}$4.15 oocytes were retrieved, with the oocyte retrieval rate per follicle of 95.0%. 3.59${\pm}$2.57 oocytes were fertilized and cleaved with the oocyte cleavage rate of 55.7%. In 83 IVF patients, 4.08${\pm}$2.39 embryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET 2.39 mebryos were transferred, and 16 pregnancies were obtained with the pregnancy rate per ET of 19.3%. In 6 GIFT patients, 7.83${\pm}$3.31 oocytes were retrieved and transferred with maximum number of 6, but no pregnancy was obtained. When compared with the previous 108 cycles of COH using FSH/hMG or pure FSH regimen, the cancellation rate during COH was significantly decreased, and all the parameters of the outcome of COH including the pregnancy rate were increased. These data suggest that GnRH agonist therapy for pituitary suppression is an effective adjunct to the current gonadotropin regimens for COH in IVF/GIFT and can increase the probability of oocytes retrieval and pregnancy, especially in the previous poor responders.

• Development and Reproduction
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• v.11 no.1
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• pp.13-20
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• 2007

In all female mammals, reproductive system is one of the first biological systems to show age-related decline. Female mammals in reproductive aging, though the phenomena is somewhat species-specific, start to show declining fertility and changes of numerous physiological functions. This review will present a current information on the aging of the female reproductive hormonal axis and introduce three useful rodent models for studying this field. Middle age($8{\sim}12$ months old) in female rats and mice is comparable to the stage prior to the entry of menopause in human. In this period pulsatile and surge GnRH secretion from hypothalamus gradually attenuated, then reduced pulsatile and surge LH secretion is followed consequently. This age-related defects in GnRH-LH neuroendocrine axis seem to be highly correlated with the defects in brain signals which modulate the activities of GnRH neuron. Many researchers support the idea which the age-related hypothalamic defects are the main cause of reproductive aging, but some ovarian factors such as inhibin response also could contribute to the induction of reproductive senescence. Some rodent models are quite valuable in studying the reproductive aging. The follitropin receptor knockout(FORKO) mice, both of null and haploinsufficient state, could produce depletion of oocyte/follicle with age. Dioxin/aryl hydrocarbon receptor(AhR) knockout mice also show severe ovarian defects and poor reproductive success early in their life compared to the age-matched normal mice. Further studies on the reproductive aging will be a great help to evaluate the benefits and risks of hormone replacement therapy(HRT) and to improve the safety of HRT.

• Korean Journal of Animal Reproduction
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• v.11 no.3
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• pp.223-229
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• 1987

This study was carried out to investigate effects of superovulation and time of embryo recovery on ovarian response, recovery rate and developmental stage of embryo in donor and effects of methods of synchronization, number of corpus luteum (CL), stage of embryo and time of embryo transfer on ovarian response, conception rate and number of newborn in recipients which were transferred on 2.5, 3.5 and 4.5 days after synchronization. The results obtained are as follows; 1. The ovulation point of superovulated donor on 2.5, 3.5 and 4.5 days after copulation was 23.3, 35.3 and 23.3, respectively. The number of embryos recovered from the donors on 2.5, 3.5 and 4.5 days after copulation was 23.3, 25.8 and 19.8, respectively. The ovulation point and number of embryos recovered on 3.5 days were greater than those of 2.5 and 4.5 days. Among 232 embryos recovered on 3.5 days after copulation, 84 were blastocyst and 62 were hatching blastocyst. 2. The number of CL in recipients on 2.5, 3.5 and 4.5 days after synchronization was 3.2, 2.9 and 3.8 and showed no difference among the days. 3. When the number of CL was 0, 2-3, 4-6 and more than 7 the pregnancy raet of recipients was 0, 37.5, 66.7 and 75%, respectively. The pregnancy rate of recipients increased as the number of CL increased. 4. The pregnancy rate of transferred morula, blastocyst and hatching blastocyst was 32.0, 37.2 and 24.7%, respectively. The blastocyst nhowed highest pregnancy rate. 5. When the recipients were synchronized by HCG, the number of CL, unruptured follicle, hemorrhage, pregnancy rate and number of young were 5.5, 6.4, 3.3, 72.7% and 3.3, whereas that of GnRH were 2.3, 4.4, 2.8, 25.0% and 1.2, respectively. Recipients synchronized by HCG showed better results than GnRH. 6. When the embryos were collected on 2.5 days after copulation and transferred to the synchromized recipients, the pregnancy rate and the number of young born was 62.58% and 3.1, respectively. Those of 3.5 and 4.5 days after copulation was 57.1% and 1.3, and 37.5% and 1.6. The 2.5 days showed higher pregnancy rate and number of young born than 3.5 and 4.5 days.

• Clinical and Experimental Reproductive Medicine
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• v.39 no.1
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• pp.22-27
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• 2012

Objective: To evaluate the effectiveness of GnRH antagonist multiple dose protocol applied during early and late follicular phase (MDP-EL) in comparison with standard GnRH agonist luteal long protocol (LP) in each non-obese and obese polycystic ovary syndrome (PCOS) women undergoing IVF. Methods: Two hundred eleven infertile women with PCOS were recruited and randomized to undergo either GnRH antagonist MDP-EL (antagonist group) or standard GnRH agonist luteal LP (agonist group). IVF cycle outcomes were compared between the two groups. Results: Total dose and days of recombinant human follicle stimulating hormone (rhFSH) administered were significantly fewer in the antagonist group than in the agonist group. Incidence of severe ovarian hyperstimulation syndrome was significantly lower in the antagonist group. However, IVF and pregnancy outcomes were similar in the two groups. When all subjects were divided into non-obese and obese subgroups, in non-obese PCOS subgroup, IVF and pregnancy outcomes were comparable in the antagonist and agonist groups but total dose and days of rhFSH were also significantly fewer in the antagonist group. Similar findings were also observed in obese PCOS subgroup. Conclusion: GnRH antagonist MDP-EL is at least as effective as GnRH agonist LP and may be a more patient-friendly alternative in controlled ovarian stimulation for PCOS patients undergoing IVF, independent of body mass index.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.12
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• pp.1695-1704
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• 2014

Maternal malnutrition during pregnancy may give rise to female offspring with disrupted ovary functions in adult age. Neonatal ovary development predisposes adult ovary function, yet the effect of maternal nutrition on the neonatal ovary has not been described. Therefore, here we show the impact of maternal protein restriction on the expression of folliculogenic and steroidogenic genes, their regulatory microRNAs and promoter DNA methylation in the ovary of neonatal piglets. Sows were fed either standard-protein (SP, 15% crude protein) or low-protein (LP, 7.5% crude protein) diets throughout gestation. Female piglets born to LP sows showed significantly decreased ovary weight relative to body weight (p<0.05) at birth, which was accompanied with an increased serum estradiol level (p<0.05). The LP piglets demonstrated higher ratio of bcl-2 associated X protein/B cell lymphoma/leukemia-2 mRNA (p<0.01), which was associated with up-regulated mRNA expression of bone morphogenic protein 4 (BMP4) (p<0.05) and proliferating cell nuclear antigen (PCNA) (p<0.05). The steroidogenic gene, cytochrome P450 aromatase (CYP19A1) was significantly down-regulated (p<0.05) in LP piglets. The alterations in ovarian gene expression were associated with a significant down-regulation of follicle-stimulating hormone receptor mRNA expression (p<0.05) in LP piglets. Moreover, three microRNAs, including miR-423-5p targeting both CYP19A1 and PCNA, miR-378 targeting CYP19A1 and miR-210 targeting BMP4, were significantly down-regulated (p<0.05) in the ovary of LP piglets. These results suggest that microRNAs are involved in mediating the effect of maternal protein restriction on ovarian function through regulating the expression of folliculogenic and steroidogenic genes in newborn piglets.

• Clinical and Experimental Reproductive Medicine
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• v.15 no.1
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• pp.40-52
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• 1988

Estradiol-17${\beta}$($E_2$) levels in the blood were estimated according to varying the time and amount of the administration of $Clomid^{(R)}$. $Clomid^{(R)}$ were administered on the 2nd, 3rd and 4th menstrual day corresponding to the recruitment period and on the 5th menstrual day corresponding to the selection period of the ovarian follicles, respectively. And $Clomid^{(R)}$ were administered 50 mg, 100 mg and 150 mg/day, repectively. The effects of the sequential HMG to $E_2$ levels in the blood were also estimated. The results were as following : 1. Blood $E_2$ levels according to the day and amount of administration of $Clomid^{(R)}$ were the highest in the group 3(D $2{\sim}6$, 150 mg/day, with HMG) and the lowest in the group 6(D $5{\sim]9$, 50 mg/day, without HMG). $E_2$ levels showed increasing tendency to 0 day. 2. In the cases of the administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the amount were similar among groups and showed increasing tendency daily. 3. In the cases of administration of $Clomid^{(R)}$ during the $2nd{\sim}6th$ menstrual day, $E_2$ levels according to the sequential HMG independent of the amount of $Clomid^{(R)}$ were higher in the with HMG group than without HMG groups. 4. In the case of the administration of $Clomid^{(R)}$ during the $5th{\sim}9th$ menstrual day, $E_2$ levels according to the amount were the highest in the 100 mg/day group and the lowest in the 50mg/day group. 5. In the cases of administration of $Clomid^{(R)}$ independent of the amount during the 5th${\sim}$9th menstrual day, $E_2$ levels according to the sequential HMG were higher in the with HMG group than without HMG group. 6. $E_2$ levels according to the amount independent of the day of the administration of $Clomid^{(R)}$ were the highest in the 100 mg/day group and 150 mg/day, 50 mg/day group in low sequence. 7. $E_2$ levels according to the sequential HMG independent of the day and amount of the administration of $Clomid^{(R)}$ were higher in the with HMG group than the without HMG group. 8. $E_2$ levels according to the day of the administration of $Clomid^{(R)}$ independent of the amount of $Clomid^{(R)}$ and sequential HMG were the highest in the group D 2${\sim}$6 and the lowest in the group D 5${\sim}$9. According to the above results, there were higher $E_2$ levels in the group with sequential HMG than without HMG. Therefore, the hypothesis, postulated initially by the author, was not verified that sequential HMG would not affect the $E_2$ levels which were related to the process of the selection of the ovarian follicle in the connection with 'FSH window'. Because it may be the stimulation after the selection of later predominant follicle. And the highest level of $E_2$ was estimated in the $Clomid^{(R)}$ 150 mg/day group with sequential HMG on the 2nd${\sim}$6th day, and the higher levels were estimated in the 2nd${\sim}$6th day, 3rd${\sim}$7th day and 4th${\sim}$8th day groups than the 5th${\sim}$9th day group. The lower levels were estimated in the $Clomid^{(R)}$ 50 mg/day group without HMG than 100 mg/day and 150 mg/day on the 5th${\sim}$9th day. Therefore, further study will be needed that combines analyses of the E2 levels in the blood according to the various administration of $Clomid^{(R)}$ with or without sequential HMG and determination of the numbers and size of the ovarian follicles by ultrasonogram.