• 한국발생생물학회지:발생과생식
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• 제4권1호
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• pp.61-66
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• 2000

본 연구에서는 조기 난소 부전증 환자를 대상으로 난포 자극 호르몬 수용체 유전자의 돌연변이와 발현 양상을 분석하였다. 돌연변이 분석을 위해 환자의 말초혈액에서 genomic DNA를 분리하고 nucleotide 566을 포함하고 있는 exon 7에 특이적인 primer쌍을 이용하여 중합효소 연쇄 반응을 시행하였다. 전기 영동으로 반응산물을 확인한 다음, 돌연변이 여부를 조사하기 위하여 제한효소 절단분석 (Restriction Fragment Length Polymorphism, RFLP)을 시행한 결과, 대조군과 조기 난소 부전증 환자군 모두에서 돌연변이를 관찰할 수 없었다. 난포 자극 호르몬 수용체의 발현양상을 확인하기 위해 시험관아기 시술과정 중 난자 채취과정에서 얻어진 황체화 과립세포에서 total RNA를 추출하여 역전사 중합효소 연쇄 반응을 시행하였다. 반응 산물을 전기 영동하여 발현양상을 비교해 본 결과, 대조군에 비해 조기 난소 부전증 환자군에서 난포 자극 호르몬 수용체 유전자의 발현이 다소 낮은 것으로 확인되었다. 이상의 결과로 보아 조기 난소 부전증 환자에서 난포 자극 호르몬 수용체 유전자의 돌연변이는 발견할 수 없었으며 난포 자극 호르몬 수용체 유전자의 발현이 대조군에 비해 낮아 과배란 유도시에 생식소 자극 호르몬에 대해 저적응증을 보이며 난포형성과정에도 장애를 받는 것으로 사료된다.

• 한국가축번식학회지
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• 제23권4호
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• pp.347-352
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• 1999

1. 유전자 재 조합 호르몬 (eCG IPMSG, hCG 및 hFSH)의 활성체크 및 세포내 signal transduction에 관한 기초연구를 위하여 rat의 융모성 성선자극 호르몬 수용체 (LH/CGR) 및 난포 자극호르몬 수용체 (FSHR)를 이미 보고되어진 염기배열에 의하여 PCR 방법으로 크로닝 하여, human embryonic kidney 유래의 293 세포에 transfection 하여 세포 표면에 LH/CGR와 FSHR를 발현하는 cell lines을 분리하였다. 2. hCG와 FSH의 signal을 전달하는 능력은 hCG와 FSH 또는 eCG의 농도증가에 따라 이들 수용체로 하여금 세포내의 cAMP 분비가 증가함을 알 수 있었다. 즉, transfection 되어진 이들 수용체를 발현하는 수용체의 대부분은 ligand binding 기능올 가지고, cAMP 반응에 의한 생리활성을 분석할 수 있으며, 또한 유전자 재조합당 단백질 호르몬 (eCG, hCG 및 hFSH)의 signal transduction, 구조 및 기능연구에 활용할 수 있다.

• Clinical and Experimental Reproductive Medicine
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• 제25권3호
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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.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2001년도 전기 제11차 학술대회 논문집
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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.

• Reproductive and Developmental Biology
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• 제42권1호
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• pp.1-6
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• 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.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2004년도 춘계학술발표대회
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• pp.209-209
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• 2004

This cDNAs were cloned with the method of the polymerase chain reaction (PCR) by sequences based on cloned rat LH/CG and FSH receptor cDNAs. A cDNAs of LHR and FSHR were transfected into the CHO cells. Several clonal cell lines were obtained expressing different numbers of cell surface receptors. (omitted)

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.340-350
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• 2007

The objective of this study was to determine whether peroxisome proliferator-activated receptor ${\gamma}$(PPAR${\gamma}$ is involved in the regulation of weaning to estrus of primiparous sows. Twelve sows composed of 6 groups of 2 full-sibs in a similar age (325.2 d), body weight (BW; 152.4 kg) and backfat thickness (BFT; 27.0 mm) at start of lactation, were allocated to accept 31 MJ (restricted group, R-group) or 53 MJ (control group, C-group) DE/d treatment, respectively. The experimental results indicated that the low energy intake resulted in excessive losses of BW and BFT during lactation in R-group sows, which may be related to decrease of serum 15-deoxy-${\Delta}^{12,14}$-prostaglandin $J_2$ (15d-$PGJ_2$), a ligand of PPAR${\gamma}$ The obvious peak and the frequency of LH, FSH and estradiol ($E_2$) were only observed in C-group sows. Except for $E_2$ at d 1 and 2, serum FSH, LH and $E_2$ concentrations in R-group were lower than those in C-group sows after weaning. However, the serum progesterone ($P_4$) level in R-group sows was always more than that in C-group. The expression abundances of PPAR${\gamma}$and GnRH receptor (GnRH-R) in pituitary, FSH receptor (FSH-R), LH receptor (LH-R), estrogen receptor (ES-R) and aromatase in ovary of anestrous sows were lower than those of estrous sows. Neither the BFT nor the BW was associated with the mRNA abundance of PPAR${\gamma}$in hypothalamus during lactation. Expressions of PPAR${\gamma}$in pituitary and ovary were affected evidently by the BFT changes and only by the loss of BW of sows during and after lactation. Furthermore, PPAR${\gamma}$mRNA level in ovary was significantly related to the expression abundances of GnRH-R, FSH-R, ES-R and aromatase, and GnRH-R was obviously associated with PPAR${\gamma}$expression in pituitary. However, PPAR${\gamma}$expression in hypothalamus likely has no effects on these genes expression and no obvious difference for all sows. Not serum $E_2$ or $P_4$ alone but the ratios of $E_2$ to $P_4$ and 15d-$PGJ_2$ to $P_4$, and serum FSH and LH were evidently related to PPAR${\gamma}$expression in pituitary and ovary. It is concluded that PPAR${\gamma}$is associated with body conditions, reproduction hormones and their receptor expression, which affected the functions of pituitary and ovary and ultimately the estrus after weaning of primiparous sows.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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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.

• Asian-Australasian Journal of Animal Sciences
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• 제16권4호
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• pp.498-503
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• 2003

Luteinizing hormone as other glycoprotein hormones is characterized by a heterodimeric structure composed a common $\alpha$-subunit noncovalently linked to a specific $\beta$-subunit. The correct conformation of the heterodimer is important for efficient secretion, hormonal-specific post-translational modifications, receptor binding and signal transduction. To determine whether $\alpha$- and $\beta$- subunits can be synthesized as a single polypeptide chain (tethered-bLH and -bFSH) and also display biological activities, the tetheredbLH and -bFSH molecules were constructed and transfected into chinese hamster ovary (CHO-K1) cells. LH and FSH activities were assayed by using the human embryonic kidney (HEK) 293 cells expressing rat LH and FSH receptor genes. The tethered-bLH and - bFSH proteins were efficiently secreted and showed a similar activity to the dimeric bovine LH and FSH $\alpha$/$\beta$ wild type and native purified from bovine pituitary. The tethered-molecules can be permit development of potent new analogues that stimulate ovarian development. Taken together, a single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds. These data indicate the potentiality of the single chain approach to further investigate structurefunction relationships of LH and FSH.

• 대한생식의학회:학술대회논문집
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• 대한불임학회 2006년도 The 5th Biannual Meeting of Pacific Rim Society for Fertility and Sterility
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• pp.163.1-163.1
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• 2006