• Reproductive and Developmental Biology
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• v.42 no.1
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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.

• Development and Reproduction
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• v.4 no.1
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• pp.87-93
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• 2000

Equine chorionic gonadotropin (eCG) consists of highly glycosylated noncovalently linked $\alpha$- and $\beta$-subunits and belongs to the glycoprotein hormone family that includes lutropin (LH), follitropin (FSH), and thyrotropin (FSH). eCG is a unique member of the gonadotropin family because it elicits response characteristics of both FSH and LH in other species than the hone. eCG is synthesized and secreted by trophoblastic cells of the endometial cups between 40 and 130 days of gestation. In the present study, mRNA expression ratio of eCG, eLH and eFSH $\alpha$- and $\beta$-subunints was investigated in the placenta and pituitary. mRNA was extracted from equine placenta on day 70 of gestation and from pituitary of male horse (27 month-old). When the expression of both subunit mRNAs of eCG in the equine placenta was compared by Northern blotting, the expression of the $\beta$ -subunit mRNA was relatively greater than that of the $\alpha$-subunit. And mRNA expression of $\alpha$-, LH $\beta$- and FSH $\beta$-subunits was analysed in the equine pituitary. An $\alpha$-subunit was revealed with a size of approximately 0.8 kb. FSH $\beta$-subunit mRNA also was detected out 1.8 kb. It is the same size of the FSH $\beta$ -subunit mRNA cloned. The intensity of $\alpha$-subunit mRNA was greater than that of the $\beta$-subunit suggesting that the expression of $\alpha$ -subunit was dominant in the equine anterior pituitary. Thus, the subunit mRNA levels seem to be independently regulated and their imbalance may account for differences in the quantities of $\alpha$- and $\beta$-subunits in the equine placenta and pituitary.

• Asian-Australasian Journal of Animal Sciences
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• v.15 no.2
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• pp.179-183
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• 2002

The fragment in intron I of FSH-${\beta}$ gene was amplified by PCR. According to the polymorphism, we analyzed the distribution of FSH-${\beta}$ retroposon in different pig breeds; its inheritance pattern in Large White${\times}$Meishan reference family; and the association of FSH-${\beta}$ retroposon with litter size, female reproductive organs measurement, ultrasonic backfat and other traits. The results showed that almost each Chinese indigenous pig had the retroposon, while foreign pig breeds rarely had; the frequencies of porcine FSH-${\beta}$ retroposon were strongly associated with breeds (p<0.01); the pattern of inheritance was consistent with Mendelian fashion; total number born (TNB) and number born alive (NBA) were increased per FSH-${\beta}$ retroposon (p<0.01) with additive effects of 1.2-1.8 and 1.4-1.8 pigs/litter, respectively; between the FSH-${\beta}$ retroposon carriers and non-carriers, there was an insignificant difference in the measurement of female reproductive organs, body weight at birth, backfat thickness, loin meat height, lean meat percentage, teat number, days to 100 kg, and average daily gain.

• Journal of Aquaculture
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• v.18 no.4
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• pp.293-298
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• 2005

We examined the effects of GnRHa on expression of the gonadotropin subunit gene in the pituitary and on syn-thesis of the plasma sex steroids (testosterone and 17$\beta$-estradiol) in protandrous black porgy. Fish were injected intraperitoneally with 0.2g GnRHa/g and then both the pituitary and the plasma were sampled 0, 6, 12, 24 and 48 hours after injection. The mRNA level of the FSH subunit increased at 6 hours post-injection, while the LH mRNA levels expressed are same with or without GnRHa treatment. Also, GnRHa stimulation caused a significant increase of the plasma testosterone (T) and 17$\beta$-estradiol ($E_2$) after 24 hours. The homologies of black porgy FSH to red seabream, Pagrus majoy FSH, snakehead fish, Channa maculata FSH and striped bass, Morone saxatilis FSH were $83.3\%,\;79.2\%$ and $76.0\%$ respectively. Amino acid homology analysis using the GenBank and EMBL general searches indicated that black porgy FSH has a high homology with yellowfin seabream, Acanthopagrus latus LH ($97.7\%$ identity) and red seabream LH ($83.3\%$ identity).

• Korean Journal of Fisheries and Aquatic Sciences
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• v.45 no.1
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• pp.17-24
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• 2012

We cloned and sequenced the cDNA encoding the $FSH{\beta}$ and $LH{\beta}$ subunits from the pituitary of the blacktip grouper Epinephelus fasciatus, which regulate vitellogenesis and maturation in vertebrates, to achieve stable and healthy gametes. The full-length cDNAs of $FSH{\beta}$ and $LH{\beta}$ were 571 bp and 617 bp, encoding 120 amino acid (aa) and 147 aa proteins, respectively. The deduced amino acid sequences of $FSH{\beta}$ and $LH{\beta}$ were highly homologous (68-97%) to those of other Perciformes; E. bruneus, Dicentrarchus labrax, Thunnus thynnus, and Pseudolabrus sieboldi. Phylogenetic analysis showed that the deduced $FSH{\beta}$ and $LH{\beta}$ amino acid sequences were categorized as a distinct subunit in the $GTH{\beta}$ family, and are closely related to the teleostei $FSH{\beta}$ and $LH{\beta}$, respectively. $FSH{\beta}$ and $LH{\beta}$ mRNA exhibited high abundance in the pituitary gland and low in other brain areas, but were not present in peripheral tissues, as determined by RT-PCR.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.49-49
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• 2003

본 연구에서는 vesicular stomatitis virus G glycoprotein (VSV-G)를 envelope로 가지는 pantropic retrovirus vector system을 이용하여 재조합 human FSH 유전자가 전이된 형질전환 닭을 생산하고자 하였다. Human FSH $\alpha$ 및 $\beta$ 유전자와 CTP linker는 human pituitary gland cDNA library에서 RT-PCR 방법을 이용하여 cloning하였으며, 각각의 fragment는 FSH$\beta$-CTP-FSH$\alpha$ 순서의 단일사슬로 연결하였다. 연결된 FSH$\beta$-CTP-FSH$\alpha$는 retroviral vector 내의 $\beta$-actin promoter의 조절 하에 도입한 후, PT67 packaging cell line에 transfection하여 virus를 생산하였으며 생산된 virus는 pantropic한 virus producing cell인 GP293에 infection하여 FSH 유전자가 도입된 virus를 생산하였다. FSH 유전자의 발현을 in vitro에서 확인하기 위하여 CHO (chinese hamster ovary) 세포에 virus를 감염시킨 후, 세포의 배양액을 취하여 electrochemilumine-scence immunoassay 방법으로 정량하였다. In vitro에서 전이 후 발현이 확인된 FSH 외래유전자의 retroviral vector virus를 초원심분리로 고농축하여 stageX의 계란의 배반엽 층에 주입하였으며, 그 결과 18%의 부화율과 91%의 부화한 닭의 유전자 전이율을 확인할 수 있었다. 전이된 유전자의 확인은 FSH$\beta$와 Neo 유전자에 대한 primer를 이용한 RT-PCR의 방법을 이용하였다. In vitro에서와는 달리 in vivo에서는 FSH 유전자의 전이는 확인되었으나 발현을 확인하지는 못하였는데, 이는 적은 수의 실험군이 형질전환율에 비해 상대적이지 못하였거나, 외래 유전자인 FSH의 발현에 의한 생리적인 부작용이 유발되어 해당개체가 부화되지 못한 것으로 추정된다. 본 문제점을 해결하기 위하여 실험군의 수를 늘리고 외래 유전자에 대한 controllable expression system이 보완될 필요성이 요구되며, 이러한 점이 해결된다면 높은 유전자 전이율에 기인하여 retrovirus를 이용한 형질전환 방법은 형질전환 가금의 생산에 있어서 매우 효율적이고 주목할 만한 방법으로 사료된다.

• Journal of Aquaculture
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• v.4 no.2
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• pp.97-110
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• 1991

This study was undertaken to understand the seasonal changes of hormone levels such as luteinizing hormone (LH), follicle stimulating hormone (FSH) and estradiol-$17{\beta}$, serum components such as albumin, total protein and triglycerides, and GSI of rainbow trout (Onco-rhynchus mykiss) under the normal-light circumstances. LH levels were the highest in November whereas FSH levels were decreased progressively from October to February. Serum GTH, such as LH and FSH, levels were low immediately after egg-stripping and were significantly increased in the summer in association with the initiation of vitellogenesis. Serum albumin, total protein, and triglycerides were significantly increased in August in association with initiation of vitellogenesis and were peak in November prior to egg-stripping. Oocyte growth, measured by oocyte diameter and GSI, was significantly increased in August and showed the highest value in January. Correlation coefficients of serum LH with FSH and estradiol-$17{\beta}$ were +0.844 and +0.947, respectively. Correlation coefficients of estradiol-$17{\beta}$ with albumin and total protein were +0.634 and +0.859, respectively. Correlation coefficients of serum estradiol-$17{\beta}$ with triglycerides and GSI were +0.673 and +0.694, respectively. Probably by positive feedback mechanism, LH, FSH and estradiol-$17{\beta}$ appeared to stimulate to liver to secrete albumin, total protein and triglycerides for vitellosenesis.

• Korean Journal of Animal Reproduction
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• v.27 no.3
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• pp.197-206
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• 2003

hFSH (human follicle stimulating hormone) is heterodimer consisting of $\alpha$ and $\beta$ subunits. Since assembly of the both subunits in the cell is often the rate-limiting step in production of functional hormone, single-chain hormones have been engineered by genetically linking two different cDNA fragments with a linker sequence. Using retrovirus vector system, the resulting recombinant hFSH gene was transferred in CHO cells and chicken embryos, and the expression of the gene was investigated. In CHO cells, protein synthesis from the single-chain FSH gene was 17 fold higher than that from the heterodimeric counterpart. In the study of transgenic chickens, ten of the eleven chicks hatched from 62 embryos manupulated with recombinant retrovirus stock was determined to carry transgenic genes. RT-PCR analyses confirmed transcription of the single-chain FSH gene, however, no recombinant FSH was detected from the blood samples.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.2
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• pp.168-173
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• 2009

Human follicle-stimulating hormone (hFSH) is a pituitary glycoprotein that regulates follicular development and ovulation. Clinically, hFSH has been used to induce follicular growth in infertile women. The hormone is composed of heterodimers, including a common ${\alpha}$ subunit among the gonadotropin family and a hormone-specific ${\beta}$ subunit. Since assembly of the heterodimer is a rate-limiting step in the production of functional hFSH, transgenic clone cows carrying a single-chain hFSH transgene may efficiently produce functional hormone. Genes encoding the ${\alpha}$ and ${\beta}$ subunits of hFSH were linked using the C-terminal peptide sequence from the ${\beta}$ subunit of human chorionic gonadotropin. Bovine fetal fibroblasts were transfected with the gene construct, including the goat ${\beta}$-casein promoter and a single-chain hFSH coding sequence. Transfected fibroblasts were transferred into enucleated oocytes, and individual nuclear transfer (NT) embryos developed to the blastocyst stage were analyzed for the transgene by polymerase chain reaction. Seventy eight blastocysts (30.8%) were developed from 259 reconstructed embryos. Among these blastocysts, the hFSH gene was detected in 70.8% (34/48) of the embryos. Subsequent transfer of hFSH-transgenic clone embryos to 31 recipients results in 11 (35.5%) early pregnancies. However, all fetuses were lost before reaching day 180 of gestation. The results from this study demonstrated that bovine NT embryos carrying single-chain hFSH could be produced, and further extensive studies in which NT embryos are transferred to more recipients may give rise to single chain hFSH-transgenic cows for biomedical applications.

• Proceedings of the Korean Society of Fisheries Technology Conference
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• 2003.05a
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• pp.248-249
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• 2003

경골어류의 뇌하수체에서는 두 종류의 생식선자극호르몬 (GTHs; FSH와 LH)이 생산된다. 이들 호르몬들은 공통적인 $\alpha$ 쇄와 특이적인 $\beta$ 쇄를 가진다 (Querat, 1995). 연어과 어종들에서, FSH는 난황형성과 정자형성의 역할을 하며, LH는 배우자의 최종성숙을 유도한다 (Blazquez et al., 1997; Swanson and Dittman, 1997). 냉수성 고유어종인 열목어의 멸종을 방지하고 번식시키기 위한 생명공학적 연구방법을 검토하기 위하여 GTH$\alpha$, FSH$\beta$와 LH$\beta$ 쇄를 발현벡터에 cloning하여 염기서열을 결정하였다. (중략)