• Title, Summary, Keyword: hCG

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Birth of a Siberian Tiger Cub from an Albino Mother Tiger with Help of eCG and hCG

  • Choo, Yoon-Jeong;Park, Myung-Soo;Han, Hyo-Dong;Ham, Gye-Sun;Park, Young-Sun;Kim, Gyeong-Sik;Park, Sun-Duk;Lim, Yang-Mook;Jung, So-Young;Yong, Hwan-Yul
    • Journal of Embryo Transfer
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    • v.26 no.3
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    • pp.215-217
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    • 2011
  • This is about the successful use of eCG and hCG for producing a Siberian tiger pup born from 10-year-old, primiparous, albino Siberian tiger. From February 2010 to July 2010, natural breeding had been tried three times with no conception. During this period of five months, estrus behaviors appeared to be typically normal and a lot of matings were observed. After consecutive failures, 1000 IU eCG (equine chorionic gonadotropin) were intramuscularly injected on the day showing estrus behavior, followed with an injection of 750 IU hCG (human chorionic gonadotropin) 80 hours later. The tiger stopped recurrence of estrus, and a cub, weighed 780 gram, was born alive 104 days after hCG injection. This study is the first report showing the unique, successful use of exogenous hormones as one of artificial breeding programs in the long history of captive breeding of carnivorous zoo animals in Korea.

hCG-induced Endoplasmic Reticulum Stress Leads to Activation of the IRE1/XBP1 Pathway in Mouse Leydig Tumor Cells (mLTC-1) (mLTC-1 세포에 hCG 처리에 의해 유도된 소포체 스트레스가 IRE1/XBP1 경로의 활성화 유발)

  • Park, Sun-Ji;Kim, Tae-Shin;Lee, Dong-Seok
    • Journal of Life Science
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    • v.24 no.10
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    • pp.1039-1045
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    • 2014
  • This study analyzed whether human chorionic gonadotropin (hCG) induces ER stress via the IRE/XBP1 pathway in mouse Leydig tumor (mLTC-1) cells. In a previous study, we demonstrated that the unfolding protein response (UPR) plays an important role in the expression of steroidogenic enzymes by modulating the ATF6 pathway, as well as ER stress-mediated apoptosis in hCG-stimulated Leydig cells. Although UPR signaling has been reported to regulate the IRE1/XBP1 pathway, it is not known whether hCG-induced ER stress in Leydig cells can activate the pathway. To investigate the activation of the IRE1/XBP1 pathway in mLTC-1 cells after hCG treatment, we performed a Western blot analysis to detect the phospho-IRE1 protein and an RT-PCR analysis to validate splicing of XBP1 mRNA. We used ER stress-activated indicator (ERAI) constructs for monitoring the activity of IRE1 and then analyzed by fluorescence microscopy and flow cytometry. The expression levels of the phospho-IRE1 protein markedly increased in response to the hCG treatment. In the mLTC-1 cells transfected with an F-XBP1-venus/F-$XBP1{\Delta}DBD$-venus construct, the hCG treatment led to the appearance of green fluorescent cells and detectable fluorescence in the nucleus and cytosol, respectively. In addition, splicing of XBP1 mRNA significantly increased after the hCG treatment. Taken together, these results indicate that hCG-induced ER stress leads to activation of the IRE1/XBP pathway in Leydig cells.

Effect of hCG on Connexin 43 mRNA Expression in Goldfish Ovary

  • Choi, Cheol-Young
    • Journal of fish pathology
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    • v.16 no.3
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    • pp.215-217
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    • 2003
  • This study examined whether the connexin (Cx) is an essential protein during oocyte maturation in the ovary of the goldfish (Carassius auratus). In mature female goldfish ovaries, at late vitellogenic stage, human insulin-like growth factor-I (IGF-I; 20 M) and human chorionic gonadotropin(hCG; 20 IU/㎖) were injected. Twelve hr after the injection, mature female goldfish ovaries were removed and stored at -80C until analysis by RT-PCR. From the goldfish Cx43 cDNA sequence (GenBank accession number AB078505), two degenerate primers were designated. In vivo, 12 hr after the treatment with hCG, goldfish Cx43 mRNA expression level was increased, while the levels of IGF-I was not changed. Goldfish Cx43 mRNA expressed after, but not before the hCG treatment. These results suggest that Cx43 mRNA was judged to be a gene, which was transcribed during oocyte maturation induced by hCG.

Assay of Human Chorionic Gonadotropin in Urine of Athletes and Evaluation of Assay Kit Performance (운동선수들의 뇨 중 Human Chorionic Gonadotropin 분석 및 분석킷트 평가)

  • 최명자;이정란;김명수
    • Biomolecules & Therapeutics
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    • v.10 no.3
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    • pp.186-192
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    • 2002
  • Special attention has been paid to human chorionic gonadotropin (hCG) for athlete doping control because it stimulates the endogenous production of testosterone and epitestosterone without increasing the T/E ratio which is a doping indicator for the exogenous administration of testosterone. Even though the IOC banned the use of hCG, a detection method has not been decided upon since there are a variety of immunoassay kits available on the market. We evaluated three kits in terms of their performance characteristics. The assay value of the control sample varied depending on the kit, resulting in 198 mIU/ml for the MAIA kit, 172mIU/ml for the IRMA kit, and 143 mIU/ml for the MEIA kit. Considering the IOC inter-lab distribution of results(55-312 mIU/ml) using 27 different kits and the mean value (178$\pm$56 mIU/ml), all three kits are within the range of -15.8% - +5.6% of the mean value, which proves them useful for the hCG assay. The MEIA kit resulted in lower hCG values because it detects only intact hCG molecules, in contrast to the other two kits which detect intact hCG and -hCG together. However, it is suitable for screening purposes because of its advantage of being an automated system. When 123 urine samples of athletes were analyzed in 22 batches using this system, the variation of control values fell within $\pm$ 10% of the mean values, and an specimens tested negative with hCG values less than the detection limit of 2 mIU/ml.

Repeated Superovulation via PMSG/hCG Administration Induces 2-Cys Peroxiredoxins Expression and Overoxidation in the Reproductive Tracts of Female Mice

  • Park, Sun-Ji;Kim, Tae-Shin;Kim, Jin-Man;Chang, Kyu-Tae;Lee, Hyun-Shik;Lee, Dong-Seok
    • Molecules and Cells
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    • v.38 no.12
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    • pp.1071-1078
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    • 2015
  • Superovulation induced by exogenous gonadotropin treatment (PMSG/hCG) increases the number of available oocytes in humans and animals. However, Superovulatory PMSG/hCG treatment is known to affect maternal environment, and these effects may result from PMSG/hCG treatment-induced oxidative stress. 2-Cys peroxiredoxins (2-Cys Prxs) act as antioxidant enzymes that protect cells from oxidative stress induced by various exogenous stimuli. Therefore, the objective of this study was to test the hypothesis that repeated PMSG/hCG treatment induces 2-Cys Prx expression and overoxidation in the reproductive tracts of female mice. Immunohistochemistry and western blotting analyses further demonstrated that, after PMSG/hCG treatment, the protein expression levels of 2-Cys Prxs increased most significantly in the ovaries, while that of Prx1 was most affected by PMSG/hCG stimulation in all tissues of the female reproductive tract. Repeated PMSG/hCG treatment eventually leads to 2-Cys Prxs overoxidation in all reproductive organs of female mice, and the abundance of the 2-Cys Prxs-$SO_{2/3}$ proteins reported here supports the hypothesis that repeated superovulation induces strong oxidative stress and damage to the female reproductive tract. Our data suggest that excessive oxidative stress caused by repeated PMSG/hCG stimulation increases 2-Cys Prxs expression and overoxidation in the female reproductive organs. Intracellular 2-Cys Prx therefore plays an important role in maintaining the reproductive organ environment of female mice upon exogenous gonadotropin treatment.

Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionic Gonadotropin Receptor

  • Min, K.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.357-364
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$ -subunits. 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 placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t631 or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632~653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agoinst-occupied receptors ~2- and ~17- fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Biological Functions of N- and O-linked Oligosaccharides of Equine Chorionic Gonadotropin and Lutropin/Chorionicgonadotropin Receptor

  • Min, K. S.
    • Proceedings of the KSAR Conference
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    • pp.10-12
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    • 2000
  • Members of the glycoprotein family, which includes CG, LH, FSH and TSH, comprise two noncovalently linked $\alpha$- and $\beta$-subunits. Equine chorionic gonadotropin (eCG), known as PMSG, has a number of interesting and unique characteristics since it appears to be a single molecule that possesses both LH- and FSH-like activities in other species than the horse. This dual activity of eCG in heterologous species is of fundamental interest to the study of the structure-function relationships of gonadotropins and their receptors. CG and LH $\beta$ genes are different in primates. In horse, however, a single gene encodes both eCG and eLH $\beta$-subunits. 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 placenta and pituitary. The dual activities of eCG could be separated by removal of the N-linked oligosaccharide on the $\alpha$-subunit Asn 56 or CTP-associated O-linked oligosaccharides. The tethered-eCG was. efficiently secreted and showed similar LH-like activity to the dimeric eCG. Interestingly, the FSH-like activity of the tethered-eCG was increased markedly in comparison with the native and wild type eCG. These results also suggest that this molecular can implay particular models of FSH-like activity not LH-like activity in the eCG/indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion. A single-chain analog can also be constructed to include additional hormone-specific bioactive generating potentially efficacious compounds that have only FSH-like activity. The LH/CG receptor (LH/CGR), a membrane glycoprotein that is present on testicular Leydig cells and ovarian theca, granulosa, luteal, and interstitial cells, plays a pivotal role in the regulation of gonadal development and function in males as well as in nonpregnant and pregnant females. The LH/CGR is a member of the family of G protein-coupled receptors and its structure is predicted to consist of a large extracellular domain connected to a bundle of seven membrane-spanning a-helices. The LH/CGR phosphorylation can be induced with a phorbol ester, but not with a calcium ionophore. The truncated form of LHR also was down-regulated normally in response to hCG stimulation. In contrast, the cell lines expressing LHR-t63I or LHR-628, the two phosphorylation-negative receptor mutant, showed a delay in the early phase of hCG-induced desensitization, a complete loss of PMA-induced desensitization, and an increase in the rate of hCG-induced receptor down-regulation. These results clearly show that residues 632-653 in the C-terminal tail of the LHR are involved in PMA-induced desensitization, hCG-induced desensitization, and hCG-induced down-regulation. Recently, constitutively activating mutations of the receptor have been identified that are associated with familial male-precocious puberty. Cells expressing LHR-D556Y bind hCG with normal affinity, exhibit a 25-fold increase in basal cAMP and respond to hCG with a normal increase in cAMP accumulation. This mutation enhances the internalization of the free and agonist-occupied receptors ~2- and ~17-fold, respectively. We conclude that the state of activation of the LHR can modulate its basal and/or agonist-stimulated internalization. Since the internalization of hCG is involved in the termination of hCG actions, we suggest that the lack of responsiveness detected in cells expressing LHR-L435R is due to the fast rate of internalization of the bound hCG. This statement is supported by the finding that hCG responsiveness is restored when the cells are lysed and signal transduction is measured in a subcellular fraction (membranes) that cannot internalize the bound hormone.

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Effects of the Administration of GnRH and HCG on the Fetus in Pregnant Rats (임신 랫드에 투여한 GnRH와 HCG가 태아에 미치는 영향)

  • 남현욱;김영홍;이근우;손창호
    • Journal of Veterinary Clinics
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    • v.20 no.2
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    • pp.212-219
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    • 2003
  • The effect of GnRH and/or hCG on the implantation, pregnancy, and the concentration of plasma estradiol and progesterone were studied in pregnant rats. GnRH 50, or 100ug and/or hCG 50 or 100 IU were administered once on day 2 or 9 of gestation, respectively. Rats were autopsied on days 8 or 16. Administration of GnRH on day 2 did not induce the prevention of implantation and termination of pregnancy but was able to induce termination of pregnancy administering on day 9. Administration of hCG induced delayed implantation on day 2 and termination of pregnancy on day 2 and 9. Administration of GnRH concomitant with hCG had no effect on prevention of implantation on day 2 but induced termination of pregnancy with a very increased fetal resorption on day 2 and with a moderate increased fetal resorption on day 9. Administration of GnRH concomitant with hCG on day 2 induced more increased termination of pregnancy compared to injection of GnRH or HCG and opposite result was observed on day 9. Plasma estradiol and progesterone concentrations by administering GnRH and/or hCG had no effect on the termination of pregnancy the pregnant rats.

Development and Immunochemical Properties of Two Monoclonal Antibodies Specific to Human Chorionic Gonadotropin

  • Kim, You-Hee;Koh, Kwan-Sam
    • BMB Reports
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    • v.32 no.5
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    • pp.474-479
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    • 1999
  • Using a hybridoma technique, spleen cells of Balb/c mice immunized with human chorionic gonadotropin (hCG) were fused with NS-1 mouse myeloma cells. Two hybrid cell lines, clones KS-8 and KS-19, secreting monoclonal antibodies to hCG, were isolated. KS-8 and KS-19 belong to the immunoglobulin $G_1$ subclass. With the aid of a double-antibody radioimmunoassay, it was established that the KS-8 monoclonal antibody recognizes an immunodeterminant of the $\beta$-subunit of hCG, whereas the KS-19 monoclonal antibody recognizes an epitope present on the $\alpha$-subunit of hCG. The KS-8 monoclonal antibody specifically reacts with human chorionic gonadotropin and shows cross-reactivity of less than 0.3% to other related human glycoprotein hormones. On the other hand, using a hemagglutination test based on antibody-induced agglutination of sheep red blood cells coated with hCG, It was shown that only the KS-19 monoclonal antibody was capable of inducing a positive reaction, although both monoclonal antibodies had similar binding capacity to the coated cells. The results from the dual screening procedures demonstrate that KS-8 and KS-19 monoclonal antibodies show high sensitivity in two different assays, and are hence useful for the qualitative and quantitative determination of hCG by both radioimmunoassay and hemagglutination inhibition tests.

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Development of recombinant human chorionic gonadotropin (hCG) using high-density culture technique of suspension-adapted chinese hamster ovary (CHO) cells

  • Na, Kyu-Heum;Kim, Seung-Chul;Seo, Kwang-Seok;Lee, Sung-Hee;Kang, Soo-Hyung
    • 한국생물공학회:학술대회논문집
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    • pp.37-37
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    • 2005
  • Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone consisting of non-covalently linked two subunits, the ${\alpha}$ and ${\beta}$ subunit. It has been used as a infertility drug for ovulation to mimic luteinizing hormone $(LH).^{1)}$ A stable cell line was established by transfection of Rc/CMV-i-dhfr-hCG, expression vector containing hCG ${\alpha}-$ and ${\beta}-genes$, into dihydrofolate reductase-deficient CHO cells and subesquent methotrexate-mediated gene amplification. Anchorage-dependent CHO cells were adapted into a serum-free and/or animal component-free suspension medium through gradual serum weaning for the hCG production. The established cell line showed typical morphological characteristics and growth profile of CHO cells, and could produce FSH with passage-to-passage consistency. The high density perfusion culture of the CHO cells was carried out in Celligen Plus bioreactor equipped with a spin-filter as a internal cell retention device. The cell density reached up to $>1x10^{7}$ cells/ml in less than 7 days and a perfusion-control strategy based on cellular consumption rates of glucose was $established.^{2)}$ Biologically active recombinant hCG was purified by a series of chromatographic steps including anion exchange chromatography and hydrophobic interaction chromatography to homogeneity. The highly purified recombinant hCG was characterized for physicochemical, immunological and biological properties.

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