• 한국수산과학회지
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• 제32권2호
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• pp.204-210
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• 1999

본 연구에서는 GTH I과 II의 분비조절기구을 밝히기 위하여 T을 경구투여한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여, activin에 의한 GTH I과 II의 분비량을 RIA로 조사하였다. 그 결과, T의 positive feedback에 의해 뇌하수체내 GTH II 함량이 증가하였으나, 뇌하수체내 GTH I 함량는 T에 의해 영향을 받지 않았다. 이러한 뇌하수체를 이용한 세포배양 실험에서, 장시간 (3 일간)의 activin 처리에 의해 GTH II 분비량은 증가하였지만, 단시간 (24시간)의 activin 처리에 의해 GTH II 분비량은 영향을 받지 않았다. 또한 activin의 자극에 의해서 분비된 GTH II 분비량은 DA에 의해 부분적으로 억제되었지만, sG-nRH의 자극에 의해서 분비된 GTH II는 DA에 의해 완전히 억제되었다. activin의 자극에 의해서 분비된 GTH II는 부분적으로 억제되었다. 그러나 activin으로 전처리에 의해 방출된 GTH II 분비량은 sGnRH 자극에 의한 증폭현상은 나타나지 않았다. 한편 GTH I 분비는 본 실험에서 사용된 호르몬에 의해서 영향을 받지 않았다. 이상의 결과들을 종합해보면, GTH I과 II는 서로 다른 합성기구에 의해 조절되며, T에 의해 GnRH, activin 그리고 DA 수용체의 감수성이 발현되어 GTH II 분비를 조절하였다. 그러나 GTH I의 분비조절 기구는 차후 계속해서 연구되어야 할 것으로 판단된다.

• 한국가축번식학회지
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• 제9권1호
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• pp.66-71
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• 1985

After the induction of estrus by the PGFF2$\alpha$-GTH combination on the 118 repeat breeder cow during of 6-13 days of the estrus cycle, the effects of GTH, antibiotics, antibiotics-GTH combination were compared. The results of this study, the following conclusion were obtained: 1. The rate of estrus was reduced the administration of GTH before the administration of PGF2$\alpha$. 2. The rate of ovulation delayed was 24.6% in the group with the administration GTH, but 34.1% without the administration GTH. Duration of ovulation was shortened by GTH. 3. The impregnation induction by the administration of PGF2$\alpha$, PGF2$\alpha$-antibiotics, PGF2$\alpha$-GTH-antibiotics were 41.7%, 69.5%, 55.6% and 80.8% respectively injection in the uterus was more effective than the administration. Before or after the A.I., the above combination was most effective. 4. In antibiotics sensitivity test in vitro of the intravagina bacteria, 5.0%, 12.7% and 60.0% of bacteria were sensitive to Penicilline, Streptomycin and Gentamicin respectively.

• Fisheries and Aquatic Sciences
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• 제2권2호
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• pp.176-181
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• 1999

The variations of gene expression and pituitary contents of GTH subunits during the ovarian development of masu salmon, Oncorhynchus masou, were investigated. The pituitary GTHs contents was measured by radioimmunoassays (RIAs) using purified GTH subunits and their antibodies. Pituitary contents of GTH $I\beta$ gradually increased from April through August, and reached the maximum in October. On the other hand, pituitary contents of GTH $II\beta$ remained low until August, but they rapidly increased in October. Total RNAs were prepared from pooled pituitaries and the GTH subunits mRNA in pituitaries was quantified by Northern blot hybridization using masu salmon cDNA probes for each GTH subunit. GTH $I\beta$ mRNA level increased with the progression of ovarian maturity. However, GTH $II\beta$ mRNA was detected only at a more advanced stage, and were extremly high at ovulation. A high levels for GTH a mRNA was detected only at ovulation stage. The synchronous increase in pituitary contents and mRNA levels suggested that ovarian maturity in masu salmon was regulated by both GTH I and GTH II.

• Fisheries and Aquatic Sciences
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• 제3권2호
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• pp.135-142
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• 2000

In order to clarify the role of gonadal sex steroids in the synthesis of gonadotropin (GTH) subunits in immature rainbow trout, we examined in vitro and in vivo effects of testosterone (T) on the pituitary mRNA levels of GTH I $\beta$, GTH II$\beta$ and a subunits by Northern blot analysis and on the pituitary content levels of GTH I$\beta$ and GTH II$\beta$by radioimmunoassay (RIA). The mRNA levels of the a subunit in T-treated fish were not changed more dramatically than those in control fish both in vivo and in vitro. Interestingly, the mRNA levels of GTH I$\beta$ in T-treated fish were shown to be slightly lower than those in the control fish under these experimental conditions, but no differences were observed in pituitary GTH I$\beta$ contents. In contrast, the mRNA levels and pituitary contents of GTH II$\beta$ subunit were strongly increased by T both in vivo and in vitro. These results demonstrate that the expressions of GTH I$\beta$ and II$\beta$ subunit genes in immatue rainbow trout pituitary are subjected to differential regulation by T.

• 한국수산과학회지
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• 제33권1호
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• pp.55-59
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• 2000

무지개 송어의 뇌하수체 및 배양액에 존재하는 GTH II 농도를 측정하기 위해 Avidin- Biotin complex를 이용한 sandwich EIA 계을 개발했다. Protein A sepharose affinity chromatography을 통해서 얻어진 연어 GTH II의 rabbit IgG에 biotinylation시킨 것 (Biotin-salmon GTH II rabbit IgG)을 제2 항체로 사용하였고, Non-Biotin salmon GTH II rabbit IgG는 단지 protein A sepharose affinity chromatography에서 얻어진 IgG를 제 1 항체로 사용하였다. EIA는 sandwich법에 의해서 이루어졌으며, 효소반응 기질로는 TMB(3,3'5,5-tetramethylbenzidine)를 이용했으며, 반응후 450 nm의 흡광도에서 automatic microplate reader로 측정하였다. 그 결과, $0.12\;{\~}\;125\;ng/ml$의 범위에서 용량반응곡선을 얻었으며, 측정감도 (최소 검출량)는 거의 0.58 ng/ml 정도 였다. 그리고 뇌하수체 추출물 및 배양액 각각의 희석곡선은 GTH II 표존곡선과 일치 하였다. 또한 이러한 GTH II의 표준곡선는 뇌하수체내 다른 peptide hormone와는 교차반응을 거의 나타내지 않았다. Testosterone을 처리한 미성숙 무지개 송어의 뇌하수체 세포배양계를 이용하여 sGnRH에 의한 GTH II 분비량을 본 sandwich EIA계와 RIA계를 비교 조사한 결과, 거의 같은 분비량을 나타냈을 뿐만아니라 같은 분비 pattern을 나타냈다. 이러한 결과로부터 본 sandwich법 EIA계에 의해서 연어과 어류의 뇌하수체 추출물 및 뇌하수체 배양액 중의 GTH II 함량 및 분비량을 측정하는데 있어서 안정된 assay계라고 생각되어진다.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.710-716
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• 2003

Gonadotropin (GTH) is a pituitary glycoprotein hormone that regulates gonadal development in vertebrates. In teleosts, two types of gonadotropins, GTH-I and GTH-II, are produced in the pituitary, and they comprised of common ${\alpha}$ and distinct ${\beta}$ subunits. In the present study, the cDNAs encoding GTH ${\alpha}\;and\;GTH-II{\beta}$ subunits were cloned and sequenced from flounder (Paralichthys olivaceus) pituitary cDNA library. The nucleotide sequence of the a subunit was 619 bp long, encoding 124 amino acids, and that of the $GTH-II{\beta}$ subunit was 538 bp long, encoding 145 amino acids. GTH subunits had well conserved cysteines, when aligned with other members of the glycoprotein family. The ${\beta}$ subunit of gonadotropin II ($GTH-II{\beta}$) had a different N-linked glycosylation site. RT-PCR analysis showed an increase of GTH II mRNA levels in association with gonadal development, and also showed that the mRNA expression of the ${\alpha}$ subunit was detected only in tissues from pituitary glands.

• Animal cells and systems
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• 제4권3호
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• pp.287-292
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• 2000

The present work examined the role of gonadotropin-releasing hormone (GnRH) and dopaminergic drugs on the secretion of maturational gonadotropin (GTH II) in relation to testosterone m treatment. This study provides evidence that the plasma GTH II levels are increased by T treatment in precocious males, but not in the immature animal. In addition, GnRH analogue (GnRHa) alone significantly increased the plasma GTH II secretion in immature rainbow trout treated with T, as well as in T-treated and T-untreated precocious males. However, injection with either dopamine (DA) or domperidone (DOM; DA D2 receptor antagonist) alone did not alter the basal plasma GTH 11 secretion in all experimental groups. The secretion of GTH II in the T-treated precocious males was remarkably influenced by GnRHa or combination of dopaminergic drugs. Notably, the effects of dopaminergic drugs on GnRHa-induced GTH II secretion w8s prolonged by T in precocious males. In T-treated immature animals, GnRHa-induced GTH II secretion was Increased only by a dose DOM (10$\mu$g/g body n) but not by higher dose DOM (100$\mu$/g body wt). In the T-untreated immature rainbow trout, however, plasma GTH 11 secretion was not influenced by the same treatments. Therefore, these results indicate that DA may be acting indirectly by blocking the effect of GnRH on GTH II secretion in vivo. T may act to modulate the relative contribution by the stimulatory (GnRH) and inhibitory (DA) neuroendocrine factors, which would ultimately determine the pattern of GTH II secretion.

• Clinical and Experimental Reproductive Medicine
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• 제16권2호
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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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• 제3권1호
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• pp.37-43
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• 2000

We used a mammalian GnRH antagonist, $[Ac-3,4-dehydro-Pro^1,\;D-p-F-Phe^2,\;D-Trp^{3.6}]$-GnRH, to examine the details of the salmon type gonadotropin-releasing hormone (sGnRH) and GnRH agonist analog $(Des-Gly^{10}$[d-Ala^6]-ethylamide GnRH; GnRHa) functions in the control of maturational gonadotropin (GTH II) secretion, in precocious male rainbow trout, in both in vivo and in vitro experiments. In the in vivo study, plasma GTH II levels increased by sGnRH or GnRHa treatment, but the response was more rapid and stronger in the GnRHa treatment group. The increase in GTH II was significantly suppressed by the GnRH antagonist, while the antagonist had no effect on basal GTH II levels in both groups. The GnRH antagonist showed stronger suppression of GTH II levels in the sGnRH treatment fish than in the GnRHa treatment fish. In addition, plasma androgenic steroid hormones (testosterone and 11-ketotestosterone) increased by the sGnRH or GnRHa treatment. The GnRH antagonist significantly inhibited the increases in plasma androgenic steroid hormone levels stimulated by the sGnRH or GnRHa, while the antagonist had no effect on basal androgenic steroid hormone levels in both groups. In the in vitro study, treatment with sGnRH or GnRHa increased GTH II release from the cultured dispersed pituitary cells, but the response was stronger in the GnRHa treatment group. The increase in GTH II release by GnRH was suppressed by adding the GnRH antagonist, dose­dependently. On the other hand, basal release of GTH II did not decrease by the GnRH antagonist treatment in both groups. These results suggest that the GnRH antagonist, $[Ac-3,4-dehydro-Pro^1,\;D-p-F-Phe^2,\;D-Trp^{3.6}]-GnRH$, used in this study is effective in blocking the action of GnRH-induced GTH II release from the pituitary gland both in vivo and in vitro.

• 한국양식학회지
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• 제9권1호
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• pp.3-10
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• 1996

인간의 태반성 성선자극호르몬(HCG) 또는 성선자극호르몬-방출호르몬 유도체(GnRH-A)와 pimozide를 이용하여 산란시기의 암컷황복에서 인공 산란을 유도하였다. 호르몬 또는 호르몬유도체의 효과는 배란 후 인공수정시켜 수정율과 배체형성율 및 면역방사측정법으로 혈액의 plasma 내 성선자극호르몬(GTH) 양을 측정하여 결정하였다. 수정율과 배체형성율이 가장 좋은 HCG 농도는 어체중 kg 당 1,000 IU이었다. 또한 GnRH-A ($10\;{\mu}g/kg$)와 pimozide를 동시에 처리했을 때 수정율과 배체형성율이 좋은 pimozide의 농도는 5 mg/kg이었다. Pimozide(1, 5 mg/kg)를 단독으로 처리한 어류에서는 plasma 내 GTH양에 영향을 줄 수 없었지만, GnRH-A와 동시에 처리했을 때 GTH양은 현저히 증가되었다. GnRH-A와 pimozide 및 dopamine을 동시에 처리했을 때 혈액의 plasma 내 GTH양은 크게 감소되었다. 이상의 결과는 산란시기 또는 이시기이외에 황복의 뇌로부터 여러 가지 성선자극호르몬과 성선자극억제호르몬을 분비됨을 시사해 준다.