• 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.

• Journal of Embryo Transfer
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• v.22 no.1
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• pp.69-74
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• 2007

This study was performed to determine the estrous cycles by macroscopic observation of the ovarian changes using the laparoscopy and to make use of these results for embryo transfer in Korean black goat (Copra hircus aegagrus). Laparoscopic examinations of the ovaries were performed from 2 days after $CIDR^(R)$ removal to 22 days after ovulation. The serial morphological changes of follicles and corpus luteum (CL) were observed. CL was classified corpus hemorrhagicum(CH), corpus luteum (CL) and corpus albicans (CA) by its maturation and regression. On the day before ovulation (Day 0), Graafian follicles (GF) were found on one or both ovaries. On the day (Day 1) and $2^{nd}$day (Day 2) of ovulation, and ovulation depression (OD) and an early stage corpus hemorrhagicum $(CH_1)$ were observed at the site of GF, respectively. On Days 3 to 4, more developed and enlarged corpus hemorrhagicum $(CH_2\;and\;CH_3)$ arised from the ovulation of the GF with well vascularization. On Days 5 to 6, it was identified that mature corpus luteum $(CL_3)$ was grown on the ovary, and fully developed CL with adjacent follicles were occupied most part of the ovary on Days 17 and 18. Then the size of CL was diminished, and completely luteal regression $(CL_1\;or\;CA)$ with new large follicle was identified on Days 20 and 22. From these results, the 4 stages of the estrous cycle in Korean black goats were 1) estrus (Day 0) for 1 day, 2) metestrus $(Day\;1{\sim}4)$ for 4 days (stage of CH development), 3) diestrus $(Day\;5{\sim}16/17)$ for 12 or 13 days (luteal stage), and 4) proestrus $(Day\;17/18{\sim}20/22)$ for 4 or 5 days (stage of luteal regression and follicular growing). Laparoscopy for observation of ovarian changes was invasive than laparotomy. Additionally, it had advantages of reduced adhesion and quick operation time. It was considered that laparoscopic examination of ovarian changes will be useful for embryo transfer in the Korean black goats.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.6
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• pp.509-519
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• 1987

Annual reproductive cycle of the Damselfish, Chromis notatus collected monthly at the four coastal areas around Chejudo, Korea are studied on the bases of histological observations of gonadal tissue and various quantitative variables including gonadosomatic index (GSI), fatness, egg diameter composition and the first maturity. The ovary consisted of a pair of saccular structure with many ovarian sacs. Oogonia proliferated along the germinal epithelium of the ovarian sac. Young oocytes with basophilic cytoplasm showed several nucleoli along the nuclear membrane. When the oocytes reached about $450{\mu}m$ in diameter, nucleus migrate toward the animal pole, nuclear membrane disappeared and most of cytoplasm were filled with yolk materials and oil drops. After ovulation, residual follicle and growing oocytes remaining in the ovarian sacs degenerated. But early young oocytes without follicle layer were not degenerated, and growing continuously till the next year. The testis consisted of a pair of lobular structures in the right and left were united in the posterior seminal vesicle. Cortex of testis was composed of many sperm ducts connected with lobuli. GSI began to increase from March, starting season of longer day length and higher water temperature, and reached the maximum value between June and August. It began to decrease from September with the lowest value appearing between October and February without any evident variation. The annual reproductive cycle could be devided into five successive stage : growing(April to Many), mature(May to August), ripe and spent (June to August) and recovery and resting stage(September to March). The spawning peak occurred from June to August. According to the frequency distribution of egg diameter, Chromis notatus was a polycyclic species to spawn twice or more in a spawning season. Fatness, correlated with gonadal phases, was remarkably decreased by spawning. Percentage of the first maturity . in femate and male fish ranging from 7.0 to 7.9 cm were $50\%$ and from 9.0 to 9.9 cm in total length $100\%$.

• Clinical and Experimental Reproductive Medicine
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• v.31 no.2
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• pp.133-139
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• 2004

Objective: To investigate the association of FSH receptor (FSHR) polymorphism at position 680 with outcomes of controlled ovarian hyper-stimulation for IVF-ET in Korean women. Design: Genetic polymorphism analysis. Materials and Methods: The FSHR polymorphism was analyzed by PCR-RFLP in 172 ovulatory women below the age of 40 year. Patients with polycystic ovary syndrome, endometriosis, or previous history of ovarian surgery were excluded. Results: Genotype distribution was 41.9% for the Asn/Asn, 47.7% for the Asn/Ser, and 10.5% for the Ser/Ser FSHR genotype group. There was no difference in age of subjects and infertility diagnosis between genotype groups. When the patients were grouped according to their FSHR genotype, the basal levels of FSH (day 3) were significantly different among the three groups ($6.0{\pm}0.3\;IU/L$ (mean $\pm$ SEM), $5.8{\pm}0.3\;IU/L$, and $8.6{\pm}1.2\;IU/L$ for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.002). The Ser/Ser group showed a higher total doses of gonadotropins required to achieve ovulation induction, and a lower serum estradiol levels at the time of hCG administration compared with other two groups, but the differences were of no statistical significance. The numbers of oocytes retrieved were significantly different among the three groups ($8.6{\pm}0.8$, $9.9{\pm}0.6$, and $6.3{\pm}0.9$, for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively, p=0.049). Clinical pregnancy rates were 42.4%, 25.9%, and 29.4% for the Asn/Asn, Asn/Ser, and Ser/Ser groups, respectively. Conclusion: Homozygous Ser/Ser genotype of FSHR polymorphism at position 680 was associated with decreased ovarian response to gonadotropin stimulation for IVF-ET.

• Korean Journal of Veterinary Research
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• v.37 no.1
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• pp.71-78
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• 1997

This study was designed to investigate the number of the growing and mature follicles following gonadotrophin treatments for superovulation in mature rats. Eighteen mature rats (Sprague-Duwely, initially 190~230gm) were randomly alloted into 3 groups. One group was control group, another FSH-treated group was injected intramuscularly with 0.5 units of follicular stimulating hormone (FSH) / rat, and third PMS and HCG-treated group was intramuscularly injected with 20~25IU of pregnant mare serum (PMS) / rat and then at the 48 hrs later, with 20~25IU of human chorionic gonadotrophin (HCG) / rat. The uteri and ovaries of rats were collected and then were observed grossly and serial sections of paraffin embedding ovaries were stained with H-E. Number of ovarian follicles by following 3 grades of large, middle and small follicles from secondary and tertiary follicles were investigated by LM photography of preparations. Small follicles were classified as secondary follicles of preantral follicles with more than 2 layers of granulosa cells surrounding the oocyte and middle follicles were classified as secondary follicles with early signs of antral cavity or with more than one small cavity on either side of the oocytes and large follicles were classified as tertiary follicles with a single medium sized antral cavity or large well-formed antral cavity. In gross findings, the uteri were slightly swelling in FSH-treated group and markedly swelling or filled with fluid in the uterine lumen in PMS and HCG-treated group. In histological findings, the shape and size of the follicles were diverse in middle and large follicles of FSH-treated group and PMS and HCG-treated group, and proportion of atretic follicles was increased in FSH-treated group and PMS and HCG-treated group than those in control group. The uteri of FSH-treated group and PMS and HCG-treated group were hypertropied or filled with fluid in the lumens and walls of uteri. The wall tissue layers were flattened and their blood and lymph vessels were dilated. The mean number of follicle per ovary in control group were appeared to be $17.1{\pm}5.6$($14.0%{\pm}4.6%$), $37.8{\pm}9.1$($30.9{\pm}7.4%$) and $67.6{\pm}30.1$($55.2{\pm}24.6%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $122.5{\pm}40.0$. The mean number of follicle per ovary in FSH-treated group were appeared to be $22.8{\pm}7.0$($17.4%{\pm}5.3%$), $43.4{\pm}6.6$($33.2{\pm}5.1%$) and $64.5{\pm}13.0$($49.3{\pm}9.9%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $130.7{\pm}16.6$. The mean number of follicle per ovary in PMS and HCG-treated group were appeared to be $29.7{\pm}11.0$($16.3%{\pm}6.0%$), $61.9{\pm}17.2$($33.9{\pm}9.4%$) and $91.1{\pm}28.2$($49.9{\pm}15.4%$) respectively at large, middle and small follicles and total number of these 3 grade follicles were appeared to be $182.6{\pm}32.7$. The above findings reveal that large follicles were increased 29.8% in FSH-treated group and 73.7% in PMS and HCG-treated group than those in control group and in histologic findings, proportion of atretic follicles were more increased in ovaries with more number of more developing follicles.

• Korean Journal of Animal Reproduction
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• v.19 no.4
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• pp.307-322
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• 1996

The corpus luteum (CL) is formed by the action of a surge of luteinizing hormone (LH) on the pre-ovulatory follicle. Luteal cells derived from granulosa and theca interna cells continue to secrete progesterone for about two weeks. LH in domestic animals is essential for the normal secretion of progesterone at all stages of the luteal phase. For this process in the rodents, 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) is indispensable. 20$\alpha$-HSD is an enzyme to be a biologically inactive steroid. This enzyme plays a critical role in the regulation of the rat luteal function and reported to be present in steroid-producing tissues such as the testis and adrenal gland. We have purified 20$\alpha$-HSD and found two distinct 20$\alpha$-HSD molecules (HSD-1 and HSD-2). Their molecular weights are both estimated to be 33kd.The amino acid compositions of HSD-1 and HSD-2 are mostly similar, but there is a slight difference in the content of lysine. We demonstrated that 1) CL of previous generations contribute more to whole ovarian 20$\alpha$-HSD activity, 2) newly formed corpora lutea contain only 20$\alpha$-HSD-1 activity, and 3) old CL express activities of each HSD isozyme as shown in the luteal tissue of cycling rats on the day of diestrus where only degenerating old CL exist. The increase in 20$\alpha$-HSD activity identified seems to be related to the increase in the numbers of 20$\alpha$-HSD-positive cells. Interestingly, 20$\alpha$-HSD-1 activities were strongly found in the follicle fluids and theca interna cells by immunohistochemical study. Thus, the activity of 20$\alpha$-HSD may be related to a survival mechanism of those luteal cells and follicles remaining in the ovaries. Luteal cells arise from two sources. The small luteal cells are all of theca cell origin, while the large luteal cells are mainly of granulosa cell origin. CL of Korean Native Cattle, as those of other animal species, contains two morphologycally and functionally distinct luteal cell populations, such as small and large luteal cells as well as nonluteal cells. In all reproductive states except in the late luteal phase, the bovine CL also contained more small luteal cells than large luteal cells. Luteal tissue secretes a variety of growth factors (proteins) and the pattern of secretion changes during all stages of the luteal phase. These growth factors could be important in regulating the function of the bovine corpus luteum and may act in a potential endocrine autocrine and paracrine mechanisms. Therefore, further work has to be done to elucidate the role of growth factors in the ovary, especially in the corpus luterum. Interest should be focussed on interaction of these growth factors in the regulation of luteal cell and the localization of cytokine synthesis in differnet luteal cells.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.69-79
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• 1989

Steroid hormone profiles during luteal phase of clomiphene citrate(CC)/human menopausal gonadotropin(hMG)/human chorionic gonadotropin(hCG)-stimulated in vitro fertilization (IVF) cycles and of follicle-stimulating hormone(FSH)/hMG/hCG-stimulated IVF cycles were compared. In seventy three cycles stimulated with CC/hMG/hCG regimen, follicles were aspirated during exploratory laparotomy and yielded 7 pregnancies, and in 83 cycles stimulated with FSH/hMG/hCG regimen, follicles were aspirated by laparoscope and made 13 pregnancies. Serum estradiol($E_2$) and progesterone($P_4$) levels were determined on days 2, 5, 7, and 9 after follicle aspiration. The FSH/hMG/hCG regimen was more effective than the CC/hMG/hCG regimen in folliculogenesis, ie, ovarian stimulation, follicular phase $E_2$ peak levels, oocyte maturation, and the number of retrieved oocytes. There was no significant difference between luteal serum $P_4/E_2$ ratio of the two regimens, suggesting that secretory endometrial build-up ability for implantation may not differ each other. Several significant correlations were observed between follicular phase seum $E_2$ peak levels and luteal phase serum $E_2$ and $P_4$ levels in the FSH/hMG/hCG-stimulated cycles but any correlation was not significant in the CC/hMG/hCG-stimulated cycles, suggesting that somewhat more follicles may eventually fall in atresia even after attaining dominant stage in the CC/hMG/hCG-stimulated cycles than the FSH/hMG/hCG-stimulated cycles.

• The Korean Journal of Zoology
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• v.33 no.2
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• pp.175-182
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• 1990

Progesterone production and oocyte maturation in ovarian follicles of Rana nigromaculata and Rana rugosa were investigated. Addition of frog pituitary homogenate (FPH) to the in utiro cultured follicles of R. nigromaculata stimulated a marked increase in the accumulation and secretion of progesterone (P$_4$) by the follicles and induced their oocyte maturation (germinal vesicle breakdown, GVBD) in a dose dependent manner. The FPH (0.1 pituitary equivalent/2 ml)-inducted P4 peak appeared in 3-6 hours and followed by the oocyte GVBD in 9-12 hours after the hormone stimulation. lncreae of intrafollicular cAMP levels with forskolin (an adenylatecyclase stimulator) and 3-isobutyl-1-methylxanthine (IBMX, a phosphodiesterase inhibitor) mimic the FPH action in the stimulation of P$_4$ production but not in the induction of oocyte maturation. The in uitro cultured follicies of R. rugosa behaved very differently from other amphibian follicles. Addition of FPH-(0. 1 pit. equivl2 ml) to the culture medium neither stimulated P$_4$ production by the follicles nor induced the oocyte GVBD. However, treatment of the follicles with forskolin and IBMX drastically stimulated both the intrafollicular accumulation (800 pg/follicle) and secretion (1700 pg/follicle) of P$_4$ by the follicles during culture period. Thus, the data suggest that the follicles are ready to respond to cAMP increase but not to the FPH stimulation in terms of P$_4$ production.

• Development and Reproduction
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• v.12 no.3
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• pp.261-266
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• 2008

In the present study, we investigated in vivo effects of Manchurian trout recombinant gonadotrophin hormone (mt-rGTH) on the induction of maturation in female eel, Anguilla japonica. The brood stock, female eel (450$\pm$50 g) were weekly injected intramuscularly with different doses of 0.1, 1, 10 ${\mu}g\m{\ell}$/fish (mt-rFSH or mt-rLH) for 10 week. The effects of r-mtGTH were analyzed by gonadosomatic index (GSI), ovarian follicle diameter and sex steroid levels. All groups did not exhibit significant differences in the GSI values. Whereas plasma testosterone (T) and estradiol-17$\beta$ (E2) levels did not change significantly in control group, plasma levels of T and E2 by injection of the r-mtFSH or r-mtLH were increased at 2 or 4 week after injection. In addition, injection of the mt-rFSH (1, 10 ${\mu}g/m{\ell}$/fish) or mt-rLH (0.1, 1, 10 ${\mu}g/m{\ell}$/fish) significantly increased follicle diameters comparing to the control group. These results demonstrate that the recombinant hormone may affect early ovary development and maturation in female eel. Taken together, these results suggest that the recombinant Manchurian trout FSH and LH are effective for reproductive activities in female eel.