• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.30-30
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• 2001

The culture of preantral follicles has important biotechnological implications through its potential to produce the large quantity of oocytes for embryo production, transgenesis research, conservation of rare breed, and a potential source of ovarian genetic material. The present study was conducted to establish the optimal conditions of in vitro culture for intact bovine preantral follicles; and to examine the developmental ability of oocytes derived from the in vitro-grown preantral follicles; and to investigate the effects of various concentrations of FSH and LH on these processes. Bovine preantral follicles (150 $\pm$ 1.2${\mu}{\textrm}{m}$), surrounded by theca cell, were isolated enzymetically and mechanically from ovarian cortical slides in Leibovitz L-15 medium containing 1 mg/$m\ell$ collagens and 0.2 mg/$m\ell$ DNase I and cultured for 25 days in the presence of different concentrations of bovine FSH and LH in $\alpha$MEM medium with insulin, transferrin, and selenite. The survival was tested by frypan Blue and Hematoxylin. The survival and growth rates of follicles were higher in FSH treatment groups than these in control (P<0.001), but there were no significant differences between the LH treatment groups and the control. In 25 days, the survival and growth rates of follicles in FSH and LH treatment group (50%, 300$\pm$1.0${\mu}{\textrm}{m}$) were higher than in FSH treatment group (40%, 244$\pm$0.5${\mu}{\textrm}{m}$) and the control group (25%, 160$\pm$ 1.0${\mu}{\textrm}{m}$). Fifty-five percent of healthy antral follicles were obtained, and 60% of the oocytes complete meiotic maturation to the metaphase II stage. Twenty-two percent of the mature oocytes underwent cleavage, and 9% developed to the blastocyst stage. In this study, in vitro-grown oocytes (111 $\pm$ $1.5mutextrm{m}$), under our culture conditions, were not equivalent in size to the in vivo-grown oocytes (130$\pm$1.3${\mu}{\textrm}{m}$). Therefore, these results suggest that bovine preantral follicles with intact theca cell can grow to the antral stage in 25days, and that oocytes from those follicles can acquire the meiotic competence and normally undergo fertilization and development to the blastocyst stage. However, the developmental capacity of in vitro-grown oocytes is presumably not comparable to those of the in vivo counterparts.

• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.27-39
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• 1984

The purpose of this experiment was to investigate the effects of experimental cryptorchidism on the levels of testicular FSH, LH receptor. One hundred and thirty two male rats weighing 100${\pm}$10g, 45 days old, were divided into three groups of control, bilateral cryptorchidism and unilateral cryptorchidism which was subdivided into two groups of scrotal testis and abdominal testis. Thirty to forty two heads of rats were allotted to each group. Five to eight rats from each group were randomely sacrificed at 1, 2, 3, 4 and 5 weeks after surgical induction of cryptorchidism. Testis was removed for assay of FSH, LH receptors. The results obtained were as follows. 1. The levels of FSH receptors per 10mg testicular tissue in abdominal testis of unilateral cryptorchid group and bilateral cryptorchid group tended to decrease gradually toward 5th week and these levels at 2nd, 3rd, 4th and 5th week were significantly lower than those levels of control group and scrotal testis of unilateral cryptorchid group. On the other hand; the ranges of affinities of FSH receptors of abdominal testis in unilateral and bilateral cryptorchid group were 2.2 - 6.67 ${\times}10^{11}M^{-1}$ and 3.6 - 4.8 ${\times}10^{11}M^{-1}$ respectively, and these values were slightly higher than those of control group (1.43 - 3.98 ${\times}10^{11}M^{-1}$) or those of scrotal testis in unilateral cryptorchid group (1.41 - 3.25 ${\times}10^{11}M^{-1}$). Especially at I and 4 weeks after treatment the affinities of abdominal testis in unilateral and bilateral cryptorchid group were significantly higher than those of other group (P <0.01). 2. The levels of LH receptors in 10mg testicular tissue tended to increase with elapse of time in control and in scrotal testis of unilateral cryptorchid group. But the levels of LH receptors of abdominal testis in unilateral and bilateral cryptorchid group tended to decrease after treatment and were significantly lower than control group at 1 (P <0.01),2,4 and 5 weeks after treatment (P < 0.05). The affinities of LH receptors were not significantly different among treatments except those of bilateral cryptorchid group and abdominal testis of unilateral cryptorchid group being significantly higher than that of the control at 4 weeks after treatment.

• Clinical and Experimental Reproductive Medicine
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• v.47 no.3
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• pp.207-212
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• 2020

Objective: Glutamate ionotropic receptor AMPA type subunit 1 (GRIA1) is a subunit of a ligand-gated ion channel that regulates the secretion of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by controlling the release of gonadotropin-releasing hormone. Few studies have investigated the association between the GRIA1 gene and human infertility. This study evaluated the association of the GRIA1 rs548294 C > T and rs2195450 G > A polymorphisms with the ovarian response to human menopausal gonadotropin (HMG) in Iranian women. Methods: One hundred women with histories of at least 1 year of infertility were included. On the second day of menstruation, patients were injected with HMG; on the third day, blood samples were collected. After hormonal analysis, the GRIA1 rs548294 C > T and rs2195450 G > A genotypes of samples were identified via the restriction fragment length polymorphism method, and on day 9, the number of follicles was assessed via ultrasound. Results: For the GRIA1 rs548294 C > T and rs2195450 G > A single nucleotide polymorphisms, the subjects with CT and GG genotypes, respectively, displayed the highest mean FSH level, LH level, and number of follicles on day 9 of the menstrual cycle (p< 0.05). Significant positive correlations were observed between LH and FSH (p< 0.01), LH and follicle count (p< 0.01), FSH and age (p< 0.05), follicle count and age (p= 0.048), and FSH and follicle count (p< 0.01). Conclusion: This study showed a significant relationship between GRIA1 polymorphisms and ovarian response to the induction of ovulation. Therefore, determining patients' GRIA1 genotype may be useful for improving treatment and prescribing suitable doses of ovulation-stimulating drugs.

• Fisheries and Aquatic Sciences
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• v.25 no.1
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• pp.12-19
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• 2022

In the present study, we investigated the effects of recombinant eel gonadotropins (rec-GTHs) on maturation induction in immature ovarian culture in vitro and sex steroid hormones in vivo in the Japanese eel Anguilla japonica. To study the in vitro effects of rec-GTHs on estradiol-17β (E2) production in immature ovarian tissues, ovarian tissues were incubated with different doses of rec-follicle-stimulating hormone (rec-FSH) or rec-luteinizing hormone (rec-LH). The results revealed that the E2 levels in the rec-FSH (0.1, 0.5, or 1 ㎍/mL)- and rec-LH (0.1 or 0.5 ㎍/mL)-treated groups were significantly higher than those in the female eels from the control group. Furthermore, to investigate the in vivo effects of rec-GTHs on the gonadosomatic index (GSI) and plasma sex steroid hormone levels, the eels were injected intraperitoneally with eel's ringer (control), salmon pituitary extract (SPE; for female eels), human chorionic gonadotropin (hCG; for male eels), rec-FSH, rec-LH, and rec-FSH + rec-LH once a week. The results revealed that except for the SPE and the hCG groups, none of the groups exhibited a significant difference in GSI values. However, in vivo plasma E2 levels increased at the end of 4 weeks after rec-FSH treatment in female eels. Based on these results, it is suggested that rec-GTHs may have a positive effect on sexual maturation in female eels; however, further studies on complementary rec-protein production systems and additional glycosylation of rec-hormones are needed to elucidate hormone bioactivity in vivo and in vitro.

• Korean Journal of Veterinary Research
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• v.36 no.4
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• pp.763-771
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• 1996

Ultrastructures of hypophyseal gonadotropes were studied in the hypophysis of the Korean native goat by electron microscopic immunohistochemistry with antisera to follicle stimulating hormone(FSH) and luteinizing hormone(LH). 1. Gonadotropes in the adenohypophysis were observed in the pars distalis and the pars tuberalis, but not in the pars intermedia. 2. FSH cells were round or oval in shape, and contained two types of round secretory granules, i.e., small and large granules. Small secretory granules were electron-dense and 180-275nm in diameter. Large secretory granules were less electron-dense than small granules and 485-555nm in diameter. 3. LH cells were round or oval in shape and also contained two types of secretory granules, the size of which were smaller than that of FSH cells. Small secretory granules were electron-dense and 160-230nm in size. The size of large secretory granules were 315-415nm with moderate electron densities.

• Journal of Embryo Transfer
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• v.33 no.4
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• pp.271-280
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• 2018

Here, we evaluated the mode of programmed cell death during porcine oocyte maturation by comparing the two major pathways associated with programmed cell death, apoptosis (type I), and autophagy (type II). We investigated the expression and localization of major genes involved in autophagy and apoptosis at mRNA and protein levels. Furthermore, the effect of hormonal stimulation on autophagy and apoptosis was analyzed. We found that the activity of autophagy-associated genes was increased in the cumulus-oocyte complexes (COCs) following follicle-stimulating hormone (FSH) treatment, while the addition of luteinizing hormone (LH) reversed this effect. The expression of proteins associated with autophagy was the highest in FSH-treated COCs. On the other hand, caspase-3 protein level was maximum in COCs cultured with LH. The treatment with rapamycin resulted in the effect similar to that observed with FSH treatment and increased autophagy activity. Thus, hormonal stimulation of pig oocytes resulted in distinct patterns of maturation. The high-quality oocytes majorly relied on the type II pathway (autophagy), while the type I pathway (apoptosis) was more prominent among poor-quality oocytes. Further investigation of this distinction may allow the development of techniques to produce high-quality oocytes in porcine in vitro fertilization.

• Reproductive and Developmental Biology
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• v.29 no.1
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• pp.25-30
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• 2005

This study was carried out to investigate the effects of semen characteristics, frozen-thawed sperm viability and serum FSH, LH, estradiol-17β and testosterone concentrations between breeds and among seasons in boars. In all seasons, Yorkshire boars produced higher semen volume compared with Duroc boars, whereas sperm concentration did not differ significantly between Duroc and Yorkshire boars. Semen volume in spring was higher compared with summer, autumn and winter in both Duroc and Yorkshire boars, but sperm concentration did not differ significantly among seasons. Sperm motility and normal acrosome rate of frozen-thawed sperm produced in spring were higher than those in summer, autumn and winter in both Duroc and Yorkshire boars. Sperm motility of frozen-thawed sperm in Yorkshire boars was higher than that in Duroc boars regardless of seasons. However, normal acrosome rate did not differ significantly between Duroc and Yorkshire boars. Serum FSH concentration in Yorkshire boars was lower than that in Duroc boars in all seasons. However, there were no significant differences on serum FSH concentration of Duroc and Yorkshire boars among seasons. Serum LH and estradiol-17β concentrations did not differ significantly between Duroc and Yorkshire boars. Also, there were no significant differences in serum LH and estradiol-17β concentrations of Duroc and Yorkshire boars among seasons. Serum testosterone concentration in Yorkshire boars was higher than that in Duroc boars in all seasons. In both breeds, serum testosterone concentrations were higher in spring than in summer, autumn and winter. In conclusion, when serum FSH concentrations were low, semen volumes were high, and when serum testosterone concentrations were high, sperm motility and normal acrosome rate of frozen-thawed sperm were high.

• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.7
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• pp.972-979
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• 2008

It has been recently shown that expression of the LH receptor (LHR) in cumulus cells is related with FSH-induced meiotic resumption of mouse cumulus enclosed oocytes (CEOs). However, to date, it is still unclear whether LHR expression in cumulus cells plays a key role during FSH-induced oocyte maturation. The purpose of this study was to characterize the functional role of LHRs in cumulus cells. CEOs were isolated from eCG-primed preovulatory follicles and cultured in hypoxanthine (HX) arrested medium. LHR protein expression in cumulus cells was time-dependent increasing during the process of FSH-induced oocyte maturation. While the sense oligodeoxynucleotide (ODN) had no effect, antisense ODN inhibited FSH-induced LHR expression and meiotic resumption. Moreover, this antisense ODN against LHR could inhibit FSH-induced mitogen-activated protein kinase (MAPK) phosphorylation. This study suggested that LHR expression in cumulus cells is involved in FSH-induced oocyte meiotic resumption, which process is possibly regulated by MAPK cascade.

• 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.