• Asian-Australasian Journal of Animal Sciences
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• v.23 no.3
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• pp.292-301
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• 2010

Follicle-stimulating hormone (FSH) is a pituitary glycoprotein hormone that is encoded by separate alpha- and betasubunit genes. It plays a key role in stimulating and regulating ovarian follicular development and egg production in chicken. FSH signal transduction is mediated by the FSH receptor (FSHR) that exclusively interacts with the beta-subunit of FSH, but characterization of prokaryotic expression of the FSHb gene and its effect on the expression of the FSHR gene in local chickens have received very little attention. In the current study, the cDNA fragment of the FSHb gene from Dagu chicken was amplified using reverse transcription polymerase chain reaction (RT-PCR), and inserted into the pET-28a (+) vector to construct the pET-28a-FSHb plasmid. After expression of the plasmid in E. coli BL21 (DE3) under inducing conditions, the recombination protein, $FSH{\beta}$ subunit, was purified and injected into the experimental hens and the effect on the mRNA expression levels of the FSHR gene was investigated. Sequence comparison showed that the coding region of the FSHb gene in the local chicken shared 99%-100% homology to published nucleotides in chickens; only one synonymous nucleotide substitution was detected in the region. The encoded amino acids were completely identical with the reported sequence, which confirmed that the sequences of the chicken FSHb gene and the peptides of the $FSH{\beta}$ subunit are highly conserved. This may be due to the critical role of the normal function of the FSHb gene in hormonal specificity and regulation of reproduction. The results of gene expression revealed that a recombinant protein with a molecular weight of about 19 kDa was efficiently expressed and it was identified by Western blotting analysis. After administration of the purified $FSH{\beta}$ protein, significantly higher expression levels were demonstrated in uterus, ovary and oviduct samples (p<0.05). These observations suggested that the expressed $FSH{\beta}$ protein possesses biological activity, and has a potential role in regulation of reproductive physiology in chickens.

• Asian-Australasian Journal of Animal Sciences
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• v.20 no.7
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• pp.1039-1048
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• 2007

Two experiments were conducted to determine whether leptin is a metabolic signal for gonadotropin secretion in ewes. In the first experiment, twenty-eight cyclic Chal ewes were assigned randomly to an energy restricted, no leptin group (ERNL) (60% of maintenance; n = 14) and an energy normal, no leptin group (ENNL) (100% of maintenance; n = 14) for 71 days (6 estrous cycles). Estrus was synchronized with seven consecutive injections of $PGF_{2{\alpha}}$ Biweekly, body weight (BW) and body condition score (BCS) were determined and blood samples were collected to measure plasma leptin concentration. Blood samples were also taken to determine plasma progesterone concentration twice weekly. After each PG injection from the second injection to the end of experiment, four ewes were selected and blood samples were collected at 20 minutes and at hourly intervals for 3 h to detect plasma LH and FSH concentration. In the second experiment, after the ceasing of the estrous cycle caused by energy restriction, six acyclic ewes were selected and randomly allotted to two groups (n = 3) and received the following treatment for four days. Ewes in an energy restricted, leptin group (ERL) were fed with a ration which provided 60% of maintenance energy requirements and intravenously injected with $4{\mu}g$ leptin/kg BW daily. Ewes in an energy excess, no leptin group (EENL) were fed with a ration that provided 180% (120%+60%) of maintenance energy requirements and intravenously injected with 1 ml saline daily. In both groups, blood samples were collected at 20 minutes and at hourly intervals for 3 h before feeding on d 0 and d 5, and for 3 h before and after injections as above on d 2 and d 4 to detect plasma LH and FSH concentration. In the first experiment, BW and BCS from the $2^{nd}$ estrous cycle, and leptin from the $3^{rd}$ estrous cycle to the end of the experiment significantly (p<0.05) decreased. In ERNL ewes, mean plasma concentrations of FSH significantly (p<0.01) decreased from the $4^{th}$ estrous cycle to d 71 and LH pulsatile secretion was suppressed on d 71, so that, mean plasma concentrations of LH (p<0.05), LH pulse frequency (p<0.01) and LH pulse amplitude (p<0.05) significantly decreased. In the second experiment, injection of leptin significantly increased mean circulating concentrations of LH (p<0.05), LH pulse frequency (p<0.01), LH pulse amplitude (p<0.05) and mean circulating concentrations of FSH (p<0.01) and leptin (p<0.01). High energy intake significantly (p<0.05) stimulated pulsatile secretion of LH and leptin secretion (p<0.01), but non-significantly increased plasma FSH concentration. The results of this study indicate that leptin is a metabolic signal for the GnRH-LH/FSH axis in feed-restricted fat-tailed ewes.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.281-286
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• 1998

Premature ovarian failure is a condition causing amenorrhea, hypoestrogenism, and elevated gonadotropins in women younger than 40 years. Many causes of premature ovarian failure were reported, including genetic abnormalities, enzymatic defects, defects in gonadotropin secretion or action, autoimmune disorders, physical and idiopathic causes. Recently, Finnish group reported a point mutation in the follicle stimulating hormone (FSH) receptor gene in premature ovarian failure patients. But it was reported that the group from United States could not find any mutation in FSH receptor gene. So we analysed C566T point mutation of FSH receptor gene using restriction fragment length polymorphism (RFLP) and compared the result between premature ovarian failure patient with idiopathic and known causes. But we did not find 556C${\rightarrow}$T mutation in the FSH receptor gene in both groups. These findings suggest that the missense mutation in the human FSH receptor gene reported in Finnish women with premature ovarian failure is uncommon in Korean women with premature ovarian failure.

• Clinical and Experimental Reproductive Medicine
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• v.20 no.2
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• pp.125-130
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• 1993

Polycystic ovarian syndrome (PCOS) patients have a characteristic of high leuteinizing hormone (LH) to follicle -stimulating hormone (FSH) ratio. Usually, human menopausal gonadotropin (hMG) is used to induce ovulation in clomiphene citrate-resistant PCOS patients. However, HMG contains two components, namely, LH and FSH, with 50%, respectively. Therefore, FSH is theoretically recommended to stimulate follicular maturation. From the pituitary, LH is secreted by pulsatile pattern. So, we have been using intermittent subcutaneous injection of pure FSH for ovulation induction in 10 PCOS patients from March, 1990 to August, 1992. We obtained good results by intermittent subcutaneous injection of pure FSH. Ovulation is 100% per patient, and 88.2% per cycle. Pregnancy rate is 80% per patient, and 23.5% per cycle. Ovarian hyperstimulation syndrome (OHSS) is 50% per patient, 41.2% per all cycles, and 46.7% per all ovulated cycles. In comparison with HMG, pregnancy rate per cycle is relatively low. But, ovulation rate and pregnacy rate per person is higher than HMG. Because of the strict check of ovaries by the vaginal ultrasonography, OHSS rate is relatively high.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2001.02a
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• pp.50-51
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• 2001

In ultrastructure study of testis, Sertoli cells start to differentiate at 16 days of gestation. Transcripts of FSH receptor, IGF-I receptor, ER $\alphareceptor, $ $TGF-\beta$ receptor and androgen receptor were highly and initially expressed at 16 day of gestation. As results of in situ PCR at 16 day of gestation, transcripts of FSH, IGF-I receptor were detected in Sertoli cells and spermatogonia, whereas the receptors of $ER\alpha, $ $TGF-\beta$ and androgen were detected in Sertoli cells. Therefore, expression of FSH and estrogen $\alpha, $ androgen, IGF-I and $TGF-\alpha$ could play an important role during fetal and prepubertal testicular development by stage specific manner in mouse.

• Korean Journal of Animal Reproduction
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• v.17 no.2
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• pp.165-171
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• 1993

While ornithine decarboxylase (ODC) is considered a key enzyme in the biosynthesis of polyamines, difluoromethylornithine(DFMO) acts as an inhibitor of polyamine synthesis. Cycling crossbred gilts were randomly assigned to one of two (treatment and control) groups (6/group). An indwelling silicone catheter was surgically implanted in the jugular vein of each animal. DFMO was dissolved in saline(200 mg/ml) and adminstered by i. m. injection at a dose of 80 mg/kg/day. The control group received an equivalent volume saline injection. DFMO was injected 3 times daily(08:00. 16:00. 24:00h) from day 16 of estrous cycle to 21 or until estrus. Once daily blood samples (10ml) were taken from day 14 until two days after the last DFMO treatment. Window blood samples were collected every 15 min for 8 h (from 08:00 to 16:00h) starting on day 16 and continuing until day 21 from one gilt per day. Serum progesterone (P$_4$), estradiol (E$_2$), LH and FSH were measured. Typical concentration profiles for P$_4$ and E$_2$ were seen during the follicular phase regardless of DFMO treatment. Injection of DFMO suppressed the preovulatory LH concentration in the serum(p<0.01) while having no effect on FSH profile. The present results indicate that DFMO had an inhibitory effect on LH secretion in the pig, but did not affect PI, E2 or FSH release.

• Clinical and Experimental Reproductive Medicine
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• v.6 no.1_2
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• pp.23-28
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• 1979

To determine whether hormone analysis of amniotic fluid could be used for accurate determination of fetal sex, we measured testosterone(T) and follicle-stimulating hormone in 19 amniotic fluid samples. The mean T in amniotic fluid of 8 women earring male fetuses was 310 pg. per milliliter and of 11 women earring female fetuses was 150 pg. per milliliter (P<0.05${\ast}$). The mean amniotic fluid FSH of 1.16 mI.U. per milliliter for 7 women with male fetuses was over trifold lower than that for subjects with female fetuses. The mean amniotic fluid FSH of female fetuses was 3.85 mI.U. per milliliter (P<0.01${\ast}$) Measurement of T & FSH in amniotic fluid may be an adjunct method for fetal sex determination.

• Journal of Embryo Transfer
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• v.17 no.1
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• pp.67-77
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• 2002

This study was carried out to assess the effect of superovulation response and quality of embryos recovered from donor cows after a single subcutaneous injection of FSH dissolved in polyethylene glycol (PEG). Cows were allocated into control and 3 experimental treatment groups. In control, cows were injected intramuscularly 50 mg FSH twice daily for 4 days. Group 1 were injected subcutaneously with a single dose of 400 mg FSH dissolved in 30% PEG solution. Group 2 were injected subcutaneously with a single dose of 200 mg FSH dissolved in 30% PEG solution. Finally in group 3, cows were injected twice 200 mg FSH dissolved in 30% PEG solution by subcutaneous. Superovulation was initiated by injection of FSH between Day 8 and 14 of the estrus cycle (Day 0, the day of estrus), and followed by injection of 25 mg PGF$_2$$\alpha$ at 48 h after first FSH injection. Cows were then artificially inseminated (AI) with semen twice at 48 and 60 h after PGF$_2$$\alpha$ injection. At 7 days after the second AI, embryos collected non-surgically by flushing the uterine horns and were counted and compared morphologically as being transferable and degenerated among different superovulation treatments. Furthermore, progesterone and estradiol -17 $\beta$ in plasma were measured by radioimmunoassay following different treatments at given days All cows of treated groups were observed heat. but control group was showed 77.8%. Superovulation response was observed as 77.8, 87.5, 88.9, and 100% in control, Groups 1, 2 and 3 The mean number of corpus lutea (CL) detected in Group 1 were 19.6, which was, respectively significantly (P<0.05) higher than those of other groups (11.1, 13.4 and 7.6, respectively). However, there did not differ on the mean number of total embryos recovered and of transferable embryos between control and treated groups.

• Clinical and Experimental Reproductive Medicine
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• v.16 no.2
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• pp.119-130
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• 1989

본 연구는 생식주기중 폐쇄여포액내에서 생물학적， 면역학적 특성을 나타내는 GTH 의 변화를 조사하고 steroid hormone과의 상관관계를 조사하며 국부조절인자로서 의 GTH의 역 활을 조사하고자 하였다. 가임기간중 215개의 여포와 IVF과정에서 185개의 여포를 얻어 여포액내 GTH의 생물학적 또는 면역학적 활성을 측정하였다. Bioactive LH(bLH)는 생쥐의 Leydig cell-testosterone production assay, bFSH는 흰쥐의 Sertoli cell aromatase assay로 측정 하였 다. Immunological GTH(iLH , iFSH) 는 MaiaClone RIA , Delfia kits를 사용하였다. 여포액내 iLH, iFSH , ihCG 는 hyperstimulation에 의해 형성된 여포의 크기와는 무관하였다. 또 hMG, huFSH 의 처리와도 상관성이 없었다. T의 농도가 높은 여포액내의 iFSH는 현저히 낮았으며 E, P 가 고농도인 여포의 ihCG 양은 현저히 낮았다. 과배란이 유도된 난소의 여포액내 iLH는 LH specific RIA로 측정시 3mIU/ml 이하이었다. 생식주기중 여포액내 bLH, bFSH는 배란기에 현저히 증가 하였다. 혈청내 GTH B/I ratio는 엘정한 반면 여포액내 LH，FSH의 생물학적， 면역학적 활성은 미수정란을 가지거나 폐쇄된 여포내의 활성보다 현저하게 높았다. 위의 결과로 보아 여포액내 생식소자극호르온은 면역학적활성보다 높은 생물학적 활성을 가지며， 생리적 현상의 지표가 된다고 추론된다. 또한 steroid, bGTH는 여포의 선택， 폐쇄를 구분하는 지표로 사용가능하며， 여포가 폐쇄될때 여포액내 B/I ratio가 현저히 낮아지는 것으로 보아 GTH의 활성이 감소되는 것으로 판단된다.

• 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.