• Clinical and Experimental Reproductive Medicine
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• v.24 no.1
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• pp.135-141
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• 1997

The early studies demonstrated that the relative amount of FSH was important for stimulating normal ovarian activity and demonstrated the existence of a threshold level for FSH, above which follicular growth was activated. It was found that only a modest increase in circulating FSH level above the threshold (between 10 and 30%) was required to stimulate folliculogenesis. In addition, FSH is primary responsible for initiating estradiol production through the activation of the aromatase enzyme system in granulosa cells, follicular secretion and growth. LH on the other hand, plays a supportive role in ovarian steroidogenesis, stimulating the ovarian thecal cells to produce androgen, the precursor for estradiol synthesis. But there is now an increasing number of reports in the literature demonstrating an adverse effect of LH on fertility and miscarriage in infertile and fertile women. So HP-FSH is the drug of a highly purified FSH preparation which has a higher specific activity and far fewer impurities than FSH. This study was performed to evaluate the efficacy and safety of HP-FSH administered (SC; subcutaneous) versus FSH(IM; intramuscular) for ovulation induction. 20 candidates patients for ovulation induction were participated. All patients underwent pituitary desensitizing with a long gonadotropin-releasing hormone (GnRH) agonist protocol and ovulation induction was started with HP-FSH SC (10 patients; group I) or FSH IM (10 patients; group II). After ovulation, outcome of ovulation induction and local reaction of injection site were compared. There were no difference of outcome of ovulation in two groups except pregnancy rate/embryo transfer. Group I had a higher pregnancy rate/ embryo transfer than Group II (44.4% Vs 28.6%). Pain, redness, tenderness, bruising and itching when the injection received on the first 5 days of treated (50 SC and 50 IM injections) were assessed. There were no significant difference (P>0.05) in the incidence of tenderness, bruising and itching between the IM and SC injection. But IM injection (FSH) had a tendency of higher above incidence. The number of reports of pain, redness were significantly increased in IM injection group (P<0.05). These results indicate that SC administration of HP-FSH has been shown to be as effect for superovulation as traditional gonadotropins, with an improved safety profile due to the removal of extaneous proteins.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.100-108
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• 1986

Eik-Nes (1966) reported that the mechanism of spermatogenesis is controlled by FSH and LH and maintaned normally in scrotum terperautre which is 3-5$^{\circ}C$ lower than body termperature. But Ojeda and Ramirez (1972) have described that the abdominal testis was shrinked severely and lost its normal function in congenital cryptorchidism or surgically induced cryptorchidism. Ramirez and Sawyer (1974) reported that the compensatory hypertorphy occured in the remaining testis of unilateral castration and the scrotal testis of unilateral cryptorchidism. Cunninham et al. (1978) reported that the serum FSH levle increased after unilateral castration. Frankel and Wright (1982) reported that the serum LH level was unchanged greatly after unilateral castration. Gomes and Jain (1976) reported that the serum testosterone level increased temporarily but not varied after unilateral castration. On the other hand, Kormano et al. (1964) reported that the serum FSH level in unilateral cryptorchidism rat was unchanged in contrast with the control and Risbirdger et al. (1981) reported that the serum LH level was unchanged till 2 weeks after operation and after then increased to 77%. Kim (1984) reported that the serum testosterone level was somewhat lower than that fo control group but there was't significant different. There were many different reports on hormone levels among different investigators when the immarue rats were castrated unilaterally or induced cryptorchidism unilaterally. Liang and Liang (1970) and Cunningham et al. (1978) described that there were no true compenastory hypertrophy in the remaining testis of unilateral castration and scrotal testis of unilateral testis of unilateral cryptorchidism in rat but they grew faster than that of control. Kormano et al.(1964), Damber et al.(1976), Cunningham et al.(1978) and Karpe et al.(1981) reported that the testis weight, germinal epithelia height and seminiferous tubules diameter developed continuously and similarily in the control, the remaining testis of unilateral castration and scrotal testis of unilateral cryptorchidism increased, however, in the abdominal testis of the unilateral cryptorchidism, they were much smaller than those of other groups. In observation of the histological changes in the seminiferous epithelium of control, remaining tesis of unilateral castration and scrotal testis of unilateral cryptorchidism differentiated and developed fully(Cunningham et al., 1978). However, the abdominal testis of unilateral crytorchidism degenerated severely and only the germ cells in early stage and Sertoli cells were found in the seminiferous tubules. (Damber et al., 1976, Gomes and Jain, 1976 and Karpe et al., 1981). By electron microscopic observation, Nagano (1963) and Leason and Leeson (1970) found that the abdominal testis of unilateral cryptorchidism was thicked in boundary tissue, increased lipid droplet in the Sertoli cell, disarranged axial filament complex and increased lipid inclusions in the Sertoli cell.

• Fisheries and Aquatic Sciences
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• v.14 no.4
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• pp.379-388
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• 2011

The mechanism by which gonadotropin-releasing hormone (GnRH) and dopamine (DA) control gonadotropin (GTH) release was studied in male and female rainbow trout using cultured pituitary cells obtained at different reproductive stages. The mechanisms of follicle-stimulating hormone (FSH) release by GnRH and DA could not be determined yet. However, basal and salmon-type GnRH (sGnRH)- or chicken-II-type GnRH (cGnRH-II)- induced luteinizing hormone (LH) release increased with gonadal maturation in both sexes. LH release activity was higher after sGnRH stimulation than cGnRH-II stimulation at maturing stages in both sexes. The GnRH antagonist ([Ac-3, 4-dehydro-$Pro^1$, D-p-F-$Phe^2$, D-$Trp^{3,6}$] GnRH) suppressed LH release by sGnRH stimulation in a dose-dependent manner, although the effect was weak in maturing fish. The role of DA as a GTH-release inhibitory factor differs during the reproductive cycle: the inhibition of sGnRH-stimulated LH release by DA was stronger in immature fish than in maturing, ovulating, or spermiated fish. DA did not completely inhibit sGnRH-stimulated LH release, and DA alone did not alter basal LH release. Relatively high doses ($10^{-6}$ or $10^{-5}M$) of domperidone (DOM, a DA D2 antagonist) increased LH release, which did not change with reproductive stage in either sex. The potency of DOM to enhance sGnRH-stimulated LH release was higher in maturing and ovulated fish than in immature fish. These data suggest that LH release from the pituitary gland is controlled by dual neuroendocrine mechanisms by GnRH and DA in rainbow trout, as has been reported in other teleosts. The mechanism of control of FSH release, however, remains unknown.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.47-53
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• 2010

Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.

• Reproductive and Developmental Biology
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• v.34 no.1
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• pp.33-40
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• 2010

Equine chorionic gonadotropin (eCG) is a heavily glycosylated glycoprotein composed of non-covalently linked $\alpha$- and $\beta$-subunits. To study the function and signal transduction of tethered recombinant-eCG (rec-eCG), a single chain eCG molecule was constructed, and the rec-eCG protein was prepared. In this study, we constructed 5 mutants (${\Delta}1$, ${\Delta}2$, ${\Delta}3$, ${\Delta}4$, and ${\Delta}5$) of rec-eCG using data about known glycoprotein hormones to analyze the role of specific follicle stimulating homone (FSH)-like activity. Three amino acids of certain specific sites were replaced with alanine. The expression vectors were transfected into CHO cells and subjected to G418 selection for 2~3 weeks. The media were collected and the quantity of secreted tethered rec-eCGs was quantified by ELISA. The LH- and FSH-like activities were assayed in terms of cAMP production by rat LH/CG and rat FSH receptors. Then, the metabolic clearance rate analyzed by the injection of rec-eCG (5 IU) into the tail vein was analyzed. The mutant eCGs (${\Delta}l$, ${\Delta}4$, and ${\Delta}5$) were transcripted, but not translated into proteins. Rec-eCG A2 was secreted in much lower amounts than the wild type. Only the rec-eCG ${\Delta}3$ ($\beta$-subunit: $Gln^{94}-Ile^{95}-Lys^{96}{\rightarrow}Ala^{94}-Ala^{95}-Ala^{96}$) was efficiently secreted. Although activity is low, its LH-like activity was similar to that of tethered $eCG{\beta\alpha}$. However, the FSH-like activity of rec-$eCG{\beta\alpha\Delta}3$ was completely flat. The result of the analysis of the metabolic clearance rate shoed the persistence of the mutant in the blood until 4 hours after the injection. After then, it almost disappeared at 8 hours. Taken together, these data suggest that 94~96 amino acid sequences in eCG $\beta$-subunit appear to be of utmost importance for signal transduction of the FSH receptor.

• Clinical and Experimental Reproductive Medicine
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• v.35 no.3
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• pp.223-229
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• 2008

Objective: This pilot study was performed to investigate the effect of metformin on insulin resistance, hormone levels, and lipid profiles in non-obese patients with polycystic ovary syndrome. Methods: This study included 16 non-obese patients with polycystic ovary syndrome diagnosed at our hospital from June 2006 to September 2007. Blood samples were collected before and 6 months after metformin treatment for analysis of fasting serum glucose levels, fasting serum insulin levels, a glycemic response to 75 g oral glucose tolerance test (OGTT), and hormonal blood profile including FSH, LH, estradiol, testosterone, free testosterone, serum lipid profiles. Insulin resistance was estimated by calculating fasting glucose/insulin ratio (FGIR), 2 hr glucose/insulin ratio after 75 g glucose load. And we investigated insulin resistance and pancreatic beta cell function by calculating HOMA beta cell function and HOMA IR. Results: After the treatment of metformin, there was significant increase in 2 hr glucose/insulin ratio after 75 g glucose load (p=0.04) and decrease in HOMA IR (p=0.000). But serum lipid profiles did not change significantly. Also the metformin treatment induced a significant reduction in serum free testosterone and LH levels, and LH/FSH ratio (p=0.001, p=0.000, p=0.034). Conclusion: This pilot study showed that metformin might be effective in improving insulin sensitivity, ameliorating hyperandrogenemia in non-obese patients with polycystic ovary syndrome. Further investigations with larger number of patients and long-term observations are necessary to determine the role of metformin.

• Clinical and Experimental Reproductive Medicine
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• v.36 no.4
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• pp.265-274
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• 2009

Objective: MicroRNAs (miR) are known to repress target genes at post-transcriptional level and play important roles in development and maturation of cell. However, the expression profiles of miR during ovarian follicle maturation have not been fully elucidated. Here, we designed this study to investigate the expression profiles of miR in oocytes and granulose cells (G-cells) after in vitro culture according to gonadotropins and adding hCG. Methods: Ovaries from 12-day-old mice (C57BL6) were removed and preantral follicles were isolated and cultured in $20\;{\mu}L$-drop of culture media with supplementation of either rFSH, rLH, or rFSH+rLH. After their full maturation, follicles were incubated with rhCG and rEGF. RNA was isolated from oocytes and G-cells, and real-time PCR were performed with primers of miR known to be expressed in the mouse ovary (mmu-miR-16, -miR-27a, -miR-126, -miR-721). Results: FSH+LH group showed the highest ovulation and MII rates among gonadotropin groups. The profiles of miRs in oocytes and G-cells differed according to gonadotropin groups and adding hCG. The profiles of miRs showed divergent changes between oocytes and G-cells. Conclusion: miR expression profiles are altered by gonadotropins and supplementation of hCG during in vitro maturation of murine follicles. Target gene study must be necessary to validate these findings.

• Clinical and Experimental Reproductive Medicine
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• v.48 no.3
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• pp.229-235
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• 2021

Objective: In this study, the effects of Lactobacillus acidophilus on testosterone (TES), follicle-stimulating hormone (FSH), luteinizing hormone (LH), androgen-binding protein (ABP), factor-associated apoptosis (FAS), and total cholesterol (TC), as well as histopathological changes, were investigated in male rats fed a high-cholesterol diet. Methods: The study included three groups. The control (C) group was fed standard-diet for 8 weeks. The hypercholesterolemia (HC) group was fed a 2% cholesterol-diet for 8 weeks. The therapeutic group (HCL) was fed a 2% cholesterol-diet for 8 weeks and administered L. acidophilus for the last 4 weeks. FSH, TES, and FAS levels in testicular tissue were determined using an enzyme-linked immunosorbent assay (ELISA), while another sample was examined histopathologically. LH and ABP levels were determined using ELISA, and serum TC levels were assessed via an autoanalyzer. Results: In the HC group, the TC levels were significantly higher and the LH levels were lower (p<0.05) than in the C group. The ABP levels were lower (p>0.05). In the HCL group, the LH and ABP levels were higher (p>0.05) and the TC level significantly lower (p<0.05) than in the HC group. The TES and FSH levels were lower, and the FAS levels were higher, in the HC than in the C group (p<0.05). In the HCL group, levels of all three resembled control levels. Histologically, in the testicular tissue of the HC group, the cells in the tubular wall exhibited atrophy, vacuolization, and reduced wall structure integrity. However, in the HCL group, these deteriorations were largely reversed. Conclusion: Supplementary dietary administration of an L. acidophilus to hypercholesterolemic male rats positively impacted testicular tissue and male fertility hormone levels.

• Reproductive and Developmental Biology
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• v.33 no.4
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• pp.229-236
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• 2009

The glycoprotein hormone family represents a class of heterodimers, that includes the placental hormone equine chorionic gonadotropin (eCG) and the anterior pituitary hormones- follitropin (FSH), lutropin (LH), and thyrotropin (TSH). The 4 hormones are heterodimers, with a common $\alpha$-subunit and unique $\beta$-subunits. eCG is the most heavily glycosylated of the known pituitary and placental glycoprotein hormones. Recent observations using single chain glycoprotein hormone analogs in which, the $\beta$-and $\alpha$-subunits are linked, implied that heterodimeric-like quaternary configuration is not a prerequisite for receptor/signal transduction. To study the function and signal transduction of tethered rec-eCG, a single chain eCG molecule was constructed and rec-eCG protein was produced. Molecular mass of the single chain is about 45 kDa. All mice were ovulated by tethered rec-eCG treatment. The dual activity of tethered rec-eCG was determined in receptor cell lines of nonequid species; in fact, this dual activity was proven in species other than horse. Tethered rec-eCG in equids does not bind to FSH receptors, suggesting that eCG is primarily an LH-like hormone in the horse. Taken together, these data suggest that tethered rec-eCG has dual activity in nonequid species in vitro. However, it has only LH-like activity in equid species in vitro.

• The Zoological Society Korea : Newsletter
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• v.12 no.2
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• pp.105.1-105
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• 1995