• Journal of Embryo Transfer
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• v.13 no.2
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• pp.159-172
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• 1998

사람 성장호르몬 수용체(hGHR) cDNA는 PCR방법에 의하여 fagment로서 보고되어진 바 있으나, liver cDNA로 부터 전장을 cloning한 보고는 없는 실정으로 본 연구에서는 기능을 가진 약 4.6kbp의 cDNA hGHR을 cloning 하는데 성공하였다. 먼저 cloning하기 위하여 human liver mRNA와 human breast cancer tissue로부터 회수한 mRNA를 RT-PCR방법에 의하여 human cDNA library와 cloning에 필요한 probe를 제작하였다. human library mRNA는 GT-PCR방법에 의하여 증폭하여 증폭되어진 산물은 λZAP Vector를 이용하여 cDNA library를 구축하였고,screeing을 위하여 임 보고 되어진 hGHR fragment native sequence를 기초로 N-terminal부분의 primer를 설계하여 950bp의 probe를 얻는데 성공하였다. 이 probe를 이용하여 준비된 human liver cDNA library로부터 2.5$\times$10 6개의 plaque로부터 6개의 positive clone을 획득하였고, 이들중 poly Asignal인 "AATAAA"를 포함하고 있는 가장 긴 약 3.8kbp의 clone을 sequencing한 결과 open reading frame을 포함하고 있었으나, 5'부분의 결손되어 있었다. 그리하여 이 부분은 human breast cancer tissue로 부터 회수한 mRNA를 RT-PCR에 의하여 증폭하였고, sequencing결과 이미 보고되어진 native hGHR와 비교한 결과 하나의 nucleotide가 silent mutation으로 판명되었다.한편 human liver cDNA library로부터 cloning한 3.8cp의 positive clone의 5'end의 결손된 부분에 silent mutation된 PCR 산물을 연결함으로써 native hGHR와 유사한 cDNA hGHR subcloning에 성공하였다. 이러한 cDNA hGHR의 clone이 function을 가지고 있는지를 검토하기 위하여 eukaryotic 발현 vector인 pCXN2에 의거 ligation한 후 chinese hamster ovary cell[CHO-KI]에 transfect를 실시하였다. Dexamethasone은 첨가하지 않고 hGH만의 존재하에서 이들 cell을 배양시키고 cell menbrane에서 발현 여부를 판정키 위하여 hGHR monocloual antibody를 사용하여 flow cytometery해석을 실시하는 한편 125I-hGH binding assay에 의하여 hGH binding activity를 측정하였다. 최종적으로 GH signal transduction의 target genedf으로 알려져 있는 serine protease inhibitor 2.1(Spi 2.1) gene의 promotor activity를 검토한 결과 hGHR을 transfect한 CHO Cell에 있어서 hGH의 농도에 의존적으로 증가되었다. 따라서 본 실험에서 cloning한 cDNA hGHR는 native hGHR와 같은 기능을 가지는 것으로 판명되었다.것으로 판명되었다.

• Korean Journal of Animal Reproduction
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• v.27 no.1
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• pp.1-7
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• 2003

Transgenic rabbits were produced by DNA microinjection using growth hormone receptor (GHR) and IGF-1 receptor (IGF-1R) genes fused to metallothionein(MT) promoter. The overall efficiencies for production of transgenic rabbits were 3.2% and 3.1% for GHR and IGF-lR genes, respectively. Founder rabbits transmitted transgenes to their progenies through medelian fashion. Growth rate of GHR and IGF-lR transgenic rabbits was significantly faster than that of non-transgenic rabbits. Transgenic rabbits grew large. (25% and 15% increase in body weight of GHR and IGF-lR transgenic rabbits, respectively) than non-transgenic rabbits and organ weight of transgenic rabbits increased, suggesting that GHR and IGF-1R genes affects growth rates in transgenic rabbits.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.4
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• pp.499-504
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• 2018

Objective: Recent studies have challenged the traditional paradigm that growth hormone receptor (GHR) displays physiological functions only in the cell membrane. It has been demonstrated that GHR localizes to the cell nucleus and still exhibits important physiological roles. The phenomenon of nuclear localization of growth hormone (GH)-induced GHR has previously been described in vitro. However, until recently, whether GH could induce nuclear localization of GHR in vivo was unclear. Methods: In the present study, we used pig as an animal model, and porcine growth hormone (pGH) or saline was injected into the inferior vena cava. We subsequently observed the localization of porcine growth hormone receptor (pGHR) using multiple techniques, including, immunoprecipitation and Western-blotting, indirect immunofluorescence assay and electronmicroscopy. Results: The results showed that pGH could induce nuclear localization of pGHR. Taken together, the results of the present study provided the first demonstration that pGHR was translocated to cell nuclei under pGH stimulation in vivo. Conclusion: Nuclear localization of pGHR induced by the in vivo pGH treatment suggests new functions and/or novel roles of nuclear pGHR, which deserve further study.

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2000.11a
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• pp.46-54
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• 2000

Transgenic rabbits were produced by DNA microinjection using growth hormone receptor (GHR) and IGF-1 receptor (IGF-1R) genes. Overall efficiencies for production of transgenic rabbits were 3.2% and 3.1% in GHR and IGF-1R genes, respectively. Founder rabbits transmitted transgenes to their progenies through medelian fashion. Growth rate in GHR and ICF-1R transgenic rabbits was faster than non-transgenic rabbits. Transgenic rabbits grew larger (25% and 15% increase in body weight of GHR and IGF-1R transgenic rabbits, respectively) than non-transgenic rabbits and organ weight of transgenic rabbits increased, suggesting that GHR and IGF-1 genes affects growth rates in transgenic rabbits.

• Korean Journal of Animal Reproduction
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• v.25 no.1
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• pp.85-90
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• 2001

To study the signaling effect of growth hormone (GH) in vivo on animal physiology, transgenic mice containing GH Receptor (GHR) gene fused to metallothionein promoter were produced by DNA microinjection into one-cell stage embryos. Three founder mice were produced with transgenic mice with approximately 4~6 copies of GHR genes and transgene was transmitted into the progeny. The founder mice were mated with normal mice to produce F$_1$ mice, and intergation and transmission of transgene were checked by polymerase chain reaction and Southern blot methods. Transmission rate of GHR transgenic mice were 20~50% in F$_1$ generation and 50% in F$_2$ generation which means that some founder mice were mosaic and transgene in F$_1$ mice was transmitted to F$_2$ progeny with Mendelian ratio. Death rate of GHR transgenic mice after birth was about 10~30% in F$_1$ and F$_2$ progenies indicating that GHR gene may affect death of transgnenic progeny.

• Asian-Australasian Journal of Animal Sciences
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• v.29 no.10
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• pp.1508-1514
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• 2016

A series of antagonists specifically targeting growth hormone receptors (GHR) in different species, such as humans, rats, bovines, and mice, have been designed; however, there are currently no antagonists that target the porcine growth hormone (GH). Therefore, in this study, we developed and characterized a porcine GHR (pGHR) antibody antagonist (denoted by AN98) via the hybridoma technique. The results from enzyme-linked immunosorbent assay, fluorescence activated cell sorter, indirect immunoinfluscent assay, and competitive receptor binding analysis showed that AN98 could specifically recognize pGHR, and further experiments indicated that AN98 could effectively inhibit pGH-induced signalling in CHO-pGHR cells and porcine hepatocytes. In addition, AN98 also inhibited GH-induced insulin-like growth factor-1 (IGF-1) secretion in porcine hepatocytes. In summary, these findings indicated that AN98, as a pGHR-specific antagonist, has potential applications in pGH-pGHR-related research on domestic pigs.

• Journal of Ginseng Research
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• v.26 no.4
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• pp.219-225
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• 2002

Study was performed to know the effects of Panax ginseng C.A. Meyer hairy root due to $^{60}$ Co ${\gamma}$-ray irradiation. We irradiated the hairy roots under the various $^{60}$ Co ${\gamma}$-ray; 0.5~4 Krad. The growth of hairy roots is inhibited over 3 Krad treatment. The lateral roots are used as a cell line after removing the apical meristem of hairy roots irradiated below 2 Krad. We selected 206 hairy root cell lines having various different growth rates and forms, and incubated in the 1/2 Murashige & Skoog(MS) medium in the absence of hormone. We selected 10 out of 206 showing superior growth. Among those, ${\gamma}$-GHR 70 and ${\gamma}$-GHR 94 showed higher growth; 34.5, 44.7%, respectively. We observed shapable, sizable characteristics according to the width of the primary roots, the process formation of the lateral roots, and the growth of lateral roots. The discriminable cell line showed that primary root is thinner, and has a vigorous growth. 8 out of 10 had much more contents than control in the aspect of the ginsenoside. ${\gamma}$-GHR 59 and ${\gamma}$-GHR 94 showed higher contents; 19, 16.9%, respectively. Therefore, we selected ${\gamma}$-GHR 70, ${\gamma}$-GHR 94 as a superior cell line in the aspect of ginsenoside contents, and growth among those irradiated by ${\gamma}$-ray. According to content of ginsenoside, Rb$_2$ effective in anticancer has 7.5% of ${\gamma}$-GHR 59. Rc, also effective in anticancer showed 16.2% content increasement of ${\gamma}$-GHR 69. It is thought that those lines will be effective in manufacturing ginsenoside. Gene analysis (VNTRP) related to the mutation is in progress.

• Korean Journal of Animal Reproduction
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• v.27 no.4
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• pp.333-338
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• 2003

Transgenic mice containing GH Receptor (GHR) gene fused to metallothionein promoter were analyzed to evaluate effect of GHR expression on growth in vivo. Three founder mice lines contained copies of GHR transgene and transmitted these genes into F$_1$ and F$_2$ progenies. The mRNA expression of transgene was identified using RT-PCR with GHR genes in tissues. To analyze the effects of transgenes on growth performance, body weights of pups were measured at 4, 10 and 14 weeks after birth. The body weight of transgenic mice was higher compared with that of non-transgenic control mice regardless of sex (P<0.05). Body weights between transgenic and non-transgenic mice were increased with aging. Overall, GHR transgenic mice tended to grow about 10 to 15 ％ faster than non-transgenic mice without any pathological defects.

• The Korean Journal of Physiology
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• v.26 no.1
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• pp.69-74
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• 1992

Central cardiovascular effects of bradykinin were examined in anesthetized normotensive (NTR) and 2-kidney, 1 clip Goldblatt hypertensive rats (GHR). Bradykinin ($0.5{\sim}10nmol$) was administered into the right lateral cerebral ventricle, while blood pressure and heart rate (HR) were continuously monitored. In both NTR and GHR, intracerebroventricular bradykinin produced a dose dependent increase in mean arterial pressure (MAP) without significant changes in HR. GHR were more sensitive in the pressor response than NTR. The pressor response to bradykinin was attenuated by treatment with hexamethonium (2.5mg/kg/min, IV) or phentolamine (2mg/kg, IV) in both NTR and GHR. Reserpine treatment (2mg/kg/day, intramuscularly,2 days) did not affect the central pressor effect of bradykinin in NTR but it attenuated the pressor effect in GHR. Pretreatment with indomethacin (10mg/kg, intraperitoneally) or saralasin ($20{\mu}g$/kg/min, IV) was without effects on the pressor response to bradykinin. These results indicate that the central pressor effect of bradykinin is, at least in part, due to excitation of the autonomic nervous activity. Mechanisms other than the enhanced sympathetic nervous activity ran. not be ruled out, However. It is also suggested that the sensitivity to bradykinin is increased in the GHR.

• Reproductive and Developmental Biology
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• v.38 no.3
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• pp.99-105
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• 2014

In this study, to further understand the mechanism of animal growth and to develop a miniature transgenic animal model, we constructed and tested tetracycline-inducible RNAi system using shRNA targeting the mRNA of mouse insulin-like growth factor (mIGF-1) or mouse growth hormone receptor (mGHR) gene. Quantitative real-time PCR analysis of mouse liver cell (Hepa1c1c7) cells transfected with these vectors showed 85% or 90% of expression inhibition effect of IGF-1 or GHR, respectively. In ELISA analysis, the protein level of IGF-1 in the cells expressing the shRNA targeting IGF-1 mRNA was reduced to 26% of non-transformed control cells. Unexpectedly, in case of using shRNA targeting GHR, the IGF-1 protein level was decreased to 75% of control cells. Further experiments are needed to explain the lower interference effect of GHR shRNA in IGF-1 protein. Accumulated knowledge of this approach could be applicable to a variety of related biological area including gene function study, gene therapy, development of miniature animals, etc.