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

• Clinical and Experimental Reproductive Medicine
• /
• v.31 no.4
• /
• pp.217-223
• /
• 2004

Objective: To evaluate the effects of recombinant FSH (rFSH) and urinary FSH (uFSH) on the gene expressions of human endometrial stromal cells in vitro. Methods: Endometrial tissue was obtained from a pre-menopausal women undergoing hysterectomy. Primary endometrial stromal cells were isolated and in vitro cultured with FBS-free DMEM/F-12 containing 0, 10, 100, and 1, 000 mIU/ml of rFSH and uFSH for 48 hours, respectively. Total RNA was extracted from the cultured cells and subjected to real time RT-PCR for the quantitative analysis of progesterone receptor (PR), estrogen receptor $\alpha/\beta$ (ER-$\alpha/\beta$), cyclooxygenase 2 (Cox-2), leukemia inhibitory factor (LIF), homeobox A10-1 and -2 (HoxA10-1/-2). Results: Both hormone treatments slightly increased (< 3 folds) the expressions of PR, ER-$\beta$ and HoxA10-1/-2 gene. However, ER-$\alpha$ expression was increased up to five folds by treatments of both FSH for 48 hours. The LIF expression by the 10 mIU/ml of uFSH for 12 hours was significantly higher than that of rFSH (p<0.01). After 24 hours treatment of two kinds of hormones, the expression patterns of LIF were similar. The 100 and 1, 000 mIU/ml of rFSH induced significantly higher amount of Cox-2 expression than those of uFSH, respectively (p<0.05). Conclusion: This study represents no adversely effect of exogeneous gonadotropins, rFSH and uFSH, on the expression of implantation related genes. We suggest that rFSH is applicable for the assisted reproductive technology without any concern on the endometrial receptivity.

• 대한생식의학회:학술대회논문집
• /
• 1996.11a
• /
• pp.67.1-67.1
• /
• 1996

• Archives of Pharmacal Research
• /
• v.25 no.3
• /
• pp.357-363
• /
• 2002

The effect of a new rhFSH, PG-0801, on oocyte quality, ovulation and in vitro fertilization (IVF) was examined in androgen-sterilized mice. Experimental sterility was induced by a single subcutaneous injection of testosterone propionate (TP, 1 mg/head) into 5 day old female mice. Ovulation was generated in the 10 to 13-week old TP-injected mice by a subcutaneous rhFSH injection (1, 5 or 10 IU/head) followed 48 hours later by a second rhFSH injection (1, 5 or 10 IU/head). For comparison, a subcutaneous PMSG (5 IU/head) injection was used for folliculogenesis and a hCG (5 IU/head) injection was used for ovulation. These were administered using the same protocol. The eggs were harvested from the oviducts and counted 17 to 20 hours after the second injection. IVF was performed by adding sperms ($2{\times}10^{5}/ml{\;}to{\;}2{\times}10^{6}/ml$) to determine the functional activity of the eggs, and the fertilization rate was measured. In addition, the pregnancy rate and fetal development were examined after 15-17 days of gestation. The number of oocytes recovered from the rhFSH/rhFSH group increased dose-dependently and was slightly higher than that of the PMSG/hCG group. The pregnancy rates of the group receiving 1, 5, and 10 IU of rhFSH/rhFSH were 50%, 66.7%, and 75%, respectively, which were significantly higher than that of the control (untreated) group (0%). The numbers of viable fetuses in the 1, 5, and 10 IU/head of the rhFSH/rhFSH group ($8.0{\pm}1.50$, $8.9{\pm}1.02$, and $8.9{\pm}1.12$ fetuses/dam, respectively) were comparable to that of the 5 IU/head PMSG/hCG group ($9.4{\pm}0.94$). The mice receiving rhFSH/rhFSH and PMSG/hCG showed similar fertilization rates (around 65%) via the IVF procedure. These results demonstrate that a new rhFSH, PG-0801, may be useful for inducing ovulation in functionally infertile patients and for superovulation in ovulatory patients participating in assisted reproductive technology (ART) programs.

• Reproductive and Developmental Biology
• /
• v.42 no.1
• /
• pp.1-6
• /
• 2018

In this study, we analyzed signal transduction by equine follicle-stimulating hormone receptor (eFSHR) on sti- mulation with recombinant $eelFSH{\beta}/{\alpha}$ ($rec-eelFSH{\beta}/{\alpha}$), natural porcine FSH (pFSH), and natural human FSH (hFSH). cAMP stimulation in CHO-K1 cells expressing eFSHR was determined upon exposure to different doses (0-1450 ng/mL) of these hormones. The $EC_{50}$ value of $rec-eelFSH{\beta}/{\alpha}$ was 53.35 ng/mL. The Rmax values of $rec-eelFSH{\beta}/{\alpha}$ and pFSH were 28.12 and 2.88 ng/mL, respectively. The activity of $rec-eelFSH{\beta}/{\alpha}$ was much higher than that of natural pFSH. However, signal transduction in CHO PathHunter Parental cells expressing eFSHR was not enhanced by stimulation with natural hFSH. Thus, $rec-eelFSH{\beta}/{\alpha}$ was completely active in cells expressing eFSHR. However, natural hFSH did not invoke a signal response in cells expressing eFSHR. Particularly, natural pFSH was weakly active in the same cells. These results showed that $eelFSH{\beta}/{\alpha}$ has potent activity in cells expressing eFSHR. Thus, $rec-eelFSH{\beta}/{\alpha}$ may efficiently bind to eFSHR, where as natural hFSH does not bind to eFSHR.

• KSBB Journal
• /
• v.22 no.2
• /
• pp.109-113
• /
• 2007

Suspension culture of recombinant Chinese hamster ovary (CHO) cells producing follicle-stimulating hormone was performed to investigate the effect of glycine betaine on cell growth and FSH production at low culture temperature. At 28$^{\circ}C$, cell growth was suppressed, but cell viability remained high for a longer culture period. When the culture temperature was lowered from 37$^{\circ}C$ to 28$^{\circ}C$, more than 14-fold increase in the maximum FSH titer was achieved. In batch culture at 28$^{\circ}C$, the use of 15 mM glycine betaine (GB) to culture medium resulted in the enhancement of maximum cell density and FSH titer by 11% and 17%, respectively, compared to the culture without GB. In pseudo-perfusion culture at 28$^{\circ}C$ with the exchange of fresh medium containing 15 mM GB, a final FSH of $2,058{\mu}g$ which is approximately 1.4-fold higher as compared to the culture without GB was obtained. This enhanced FSH production with 15 mM GB was not just because of enhanced specific FSH productivity (qFSH), but mainly because of the extended culture longevity. Taken together, this result demonstrates that the application of GB at low culture temperature is feasible to enhance the production of recombinant proteins in rCHO cells.

• Development and Reproduction
• /
• v.4 no.1
• /
• pp.13-17
• /
• 2000

Follicle stimulating hormone (FSH) stimulates follicle growth, and inhibits the follicle atresia in the immature rodent ovaries. The present study was carried out to know the histological changes of ovarian follicles after FSH treatment in the prepubertal mice. Ten i.u. of recombinant FSH was i.p. injected on 3 weeks old mice. After the treatment, at 1, 2 and 3 days, left ovaries were collected for the histological study. The atretic ratio of preantral follicles increased with time after FSH treatment. However, in the case of antral follicles, there was no significant change in the ratio. The degenerating follicles contained apoptotic granulosa cells, macrophage, and polymorphonuclear leukocytes in the follicular cavity. The present results suggest that follicular degeneration caused by FSH hyperstimulation could be mediated by apoptosis as well as the acute inflammation.

• 대한생식의학회:학술대회논문집
• /
• 1999.11a
• /
• pp.31-32
• /
• 1999

• Korean Journal of Animal Reproduction
• /
• v.26 no.4
• /
• pp.395-399
• /
• 2002

Human chorionic gonadotropin (hCG) is a member of the glycoprotein hormone family which includes FSH. hCG TSH. These hormone family is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a hormone specific $\beta$-subunit. To determine u and $\beta$ -subunits can be synthesized as a single polypeptide chain (tethered-hCG) and also display biological activity, the tethered-hCC and -FSH molecule by fusing the carboxyl terminus of the hCG $\beta$-subunit to the amino terminus of the $\alpha$-subunit was constructed. To determine the importance of $\alpha$ COOH -terminal amino acid, we also deleted the $\alpha$ COOH-terminal amino acids. The expressing vectors were transfected into CHO-K 1 cells. The tethered-wthCG and -wtFSH was efficiently secreted. The $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants had no secretion. These results are the first conclusive evidence that COOH-terminal amino acids are very important for secretion in human glycoprotein hormone $\alpha$-subunit. These results demonstrated that the $\alpha$ Δ83hCG and $\alpha$ Δ 83FSH mutants could be play a pivotal role in the secretion of tethered-molecule.

• 대한생식의학회:학술대회논문집
• /
• 2002.05a
• /
• pp.65-66
• /
• 2002