• Korean Journal of Agricultural Science
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• v.12 no.1
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• pp.70-74
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• 1985

Serum levels of LH, FSH, prolactin, estradiol-$17{\beta}$ and progesterone were investigated every 20 days after the day of mating for the period of 140 days, at the day of parturition and thereafter 10 th and 20th day postpartum in Korean native goats. Serum levels of LH were highest with 1.95 mIU/ml at the 60 days after gestation, but maintained high levels until 100 days after gestation, and thereafter decreased gradually to 0.02 mIU/ml at the day of parturition. Prolactin concentrations increased from the 140 days after gestation, and showed highest levels with 29.75 ng/ml at the day of parturition, then decreased gradually. FSH levels determined were below 1.25 mIU/ml during the experimental period. Estradiol-$17{\beta}$ concentrations increased gradually from 7.51 pg/ml at the 20 days after gestation to 159.62 pg/ml at the day of parturition, and decreased rapidly after parturition. Progesterone levels were highest with 6.62 ng/ml at the 120 days after gestation, the decreased rapidly to 1.25 ng/ml at the day of parturition.

• Development and Reproduction
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• v.13 no.4
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• pp.257-264
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• 2009

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD) due to its specific toxicity against dopaminergic (DA) neurons. Since DA signals modulate a broad spectrum of CNS physiology, one can expect profound alterations in neuroendocrine activities of both PD patients and 6-OHDA treated animals. Limited applications of 6-OHDA injection model, however, have been made on the studies of hypothalamuspituitary neuroendocrine circuits. The present study was performed to examine whether blockade of brain catecholamine (CA) biosynthesis with 6-OHDA can make any alteration in the transcriptional activities of hypothalamus-pituitary hormone genes in adult male rats. Three-month-old male rats (SD strain) were received 6-OHDA ($200{\mu}g$ in $10{\mu}\ell$ of saline/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. To determine the mRNA levels of hypothalamuspituitary hormone genes, total RNAs were extracted and applied to the semi-quantitative RT-PCRs. The mRNA levels of tyrosine hydroxylase (TH), the rate-limiting enzyme for the catecholamine biosynthesis, were significantly lower than those from the control group (control:6-OHDA=1:0.72${\pm}$0.02AU, p<0.001), confirming the efficacy of 6-OHDA injection. The mRNA levels of gonadotropin-releasing hormone (GnRH) and corticotropin releasing hormone (CRH) in the hypothalami from 6-OHDA group were significantly lower than those from the control group (GnRH, control:6-OHDA=1:0.39${\pm}$0.03AU, p<0.001; CRH, control:6-OHDA=1:0.76${\pm}$0.07AU, p<0.01). There were significant decreases in the mRNA levels of common alpha subunit of glycoprotein homones (Cg$\alpha$), LH beta subunit (LH-$\beta$), and FSH beta subunit (FSH-$\beta$) in pituitaries from 6-OHDA group compared to control values (Cg$\alpha$, control:6-OHDA=1:0.81${\pm}$0.02AU, p<0.001; LH-$\beta$, control:6-OHDA=1:0.68${\pm}$0.04AU, p<0.001; FSH-$\beta$, control:6-OHDA=1:0.84${\pm}$0.05AU, p<0.001). Similarly, the level of adrenocorticotrophic hormone (ACTH) transcripts from 6-OHDA group was significantly lower than that from the control group (control: 6-OHDA=1:0.86${\pm}$0.04AU, p<0.01). The present study demonstrated that centrally injected DA neurotoxin could downregulate the transcriptional activities of the two hypothalamus-pituitary neuroendocrine circuits, i.e., GnRH-gonadotropins and CRH-ACTH systems. These results suggested that hypothalamic CA input might affect on the activities of gonad and adrenal through modulation of hypothalamus-pituitary function, providing plausible explanation for frequent occurrence of sexual dysfunction and poor stress-response in PD patients.

• Development and Reproduction
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• v.21 no.2
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• pp.111-120
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• 2017

Equine chorionic gonadotropin (eCG) is a unique molecule that elicits the response characteristics of both follicle-stimulating hormone (FSH) and luteinizing hormone (LH) in other species. Previous studies from this laboratory had demonstrated that recombinant eCG (rec-eCG) from Chinese hamster ovary (CHO-K1) cells exhibited both FSH- and LH-like activity in rat granulosa and Leydig cells. In this study, we analyzed receptor internalization through rec-eCGs, wild type eCG ($eCG{\beta}/{\alpha}$) and mutant eCG ($eCG{\beta}/{\alpha}{\Delta}56$) with an N-linked oligosaccharide at $Asn^{56}$ of the ${\alpha}-subunit$. Both the rec-eCGs were obtained from CHO-K1 cells. The agonist activation of receptors was analyzed by measuring stimulation time and concentrations of rec-eCGs. Internalization values in the stably selected rat follicle-stimulating hormone receptor (rFSHR) and rat luteinizing/chorionic gonadotropin receptor (rLH/CGR) were highest at 50 min after stimulation with 10 ng of $rec-eCG{\beta}/{\alpha}$. The dose-dependent response was highest when 10 ng of $rec-eCG{\beta}/{\alpha}$ was used. The deglycosylated $eCG{\beta}/{\alpha}{\Delta}56$ mutant did not enhance the agonist-stimulated internalization. We concluded that the state of activation of rFSHR and rLH/CGR could be modulated through agonist-stimulated internalization. Our results suggested that the eLH/CGRs are mostly internalized within 60 min by agonist-stimulation by rec-eCG. We also suggested that the lack of responsiveness of the deglycosylated $eCG{\beta}/{\alpha}{\Delta}56$ was likely because the site of glycosylation played a pivotal role in agonist-stimulated internalization in cells expressing rFSHR and rLH/CGR.

• 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.

• 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.

• Animal cells and systems
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• v.16 no.2
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• pp.127-134
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• 2012

We investigated the effects of fadrozol, an aromatase inhibitor (AI), and $17{\alpha}$-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper $Epinephelus$ $bruneus$, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-${\beta}$ and LH-${\beta}$) in the pituitary, and estradiol-$17{\beta}$ (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers ($113{\pm}17g\;body\;weight$; $16.2{\pm}1.2cm\;body\;length$) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BWdoses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-${\beta}$ mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-${\beta}$ mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-${\beta}$.

• 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.

• 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.

• 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.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.125-142
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• 2000

13). Analysis of a purified preparation of eCG revealed that its $\beta$ -subunit consists of 149 amino acids, which was confirmed by the molecular cloning of its cDNA. There seem to be at least four to six, or even as many as 11, O-glycosylation sites on the extended C-tenninal region of the eCG $\beta$-subunit. Interestingly, eCG is a unique member of this family, as it appear to be a single molecule that possesses both LH- and FSH-like activities. Using the cDNA prepared from mRNA extracted from equine placental and pituitary tissues, we cloned the cDNA of eCG $\alpha$- and $\beta$ -subunits and eFSH $\beta$ -subunit. The mRNA expression of each subunit seems to be independently regulated, which may account for differences in the quantities of $\alpha$ - and $\beta$ -subunits in the placenta and pituitary. Thus, eCG is a distinct molecule from the view points of its biological function and glycoresidue structures. Recombinant eCGs including the mutants which lack oligosaccharides will be useful tools for analyzing the structure-function relationships of gonadotropins in the horse as well as other species. Similar experiments will also clarify the proposed structure and biological functions for the glycoprotein hormones. These experimental are now possible, and hopefully a resolution of the existing controversy will be forthcoming in the near future.