DOI QR코드

DOI QR Code

Signal Transduction of Equine Follicle-Stimulating Hormone Receptor (eFSHR) by rec-eelFSHβ/α, Natural Porcine FSH, and Natural Human FSH

  • Byambaragchaa, Munkhzaya (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Life Science, Institute of Genetic Engineering, Hankyong National University) ;
  • Kim, Dae-Jung (Aquaculture Research Division, National Institute of Fisher Science (NIFS)) ;
  • Kang, Myung-Hwa (Department of Food Science and Nutrition, Hoseo University) ;
  • Min, Kwan-Sik (Animal Biotechnology, Graduate School of Future Convergence Technology, Department of Animal Life Science, Institute of Genetic Engineering, Hankyong National University)
  • Received : 2018.02.23
  • Accepted : 2018.03.02
  • Published : 2018.02.28

Abstract

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.

Keywords

References

  1. Guillou F, Combarnous Y (1983): Purification of equine gonadotropins and comparative study of their aciddissociation and receptor-binding specificity. Biochim Biophys Acta 755:229-236. https://doi.org/10.1016/0304-4165(83)90208-8
  2. Jeoung YH, Yoon JT, Min KS (2010): Biological functions of the COOH-terminal amino acids of the $\alpha$-subunit of tethered equine chorionic gonadotropin. Reprod Dev Sci 34:47-53.
  3. Kagawa H, Iinuma N, Tanaka H, Ohta H, Okuzawa K (1998): Effects of rearing period in seawater on induced maturation in female Japanese eel Anguilla japonica. Fish Sci 64:77-82. https://doi.org/10.2331/fishsci.64.77
  4. Kazeto Y, Kohara M, Miura T, Miura C, Yamaguchi S, Trant JM, Adachi S, Yamauchi K (2008): Japanese eel follicle-stimulating hormone and luteinizing hormone:production of biologically active recombinant FSH and LH by drosophila S2 cells and their differential actions on the reproductive biology. Biol Reprod 79:938-946. https://doi.org/10.1095/biolreprod.108.070052
  5. Kazeto Y, Ozaki Y, Ito R, Suzuki H, Tanaka T, Imaizumi H, Nomura K, Okuzawa K, Gen K (2014): Mass production of recombinant Japanese eel follicle-stimulating hormone and luteinizing hormone: their differential actions on gametogenesis in vitro. In: IC-FCEN 2014). University of Hyderavad, India, pp. 17.
  6. Kim DJ, Bae JY, Kim EO (2007): Changes in sex steroid hormones and ovarian development during artificial maturation of female eel. Anguilla japonica. Integ Biosci 11:117-124. https://doi.org/10.1080/17386357.2007.9647323
  7. Kim DJ, Park CW, Byambaragchaa M, Kim SK, Lee BI, Hwang HK, Myeong JI, Hong SM, Kang MH, Min KS (2016a): Data on the characterization of follicle-stimulating hormone monoclonal antibodies and localization in Japanese eel pituitary. Data in Brief 8: 404-410. https://doi.org/10.1016/j.dib.2016.05.069
  8. Kim DJ, Park CW, Kim DW, Park HK, Byambaragchaa M, Lee NS, Hong SM, Seo MY, Kang MH, Min KS (2016b): Production and characterization of monoclonal antibodies against recombinant tethered follicle-stimulating hormone from Japanese eel Anguilla japonica. Gem Comp Endocrinol 233:8-15. https://doi.org/10.1016/j.ygcen.2016.04.030
  9. Kobayashi M, Hayakawa Y, Park W, Banba A, Yoshizaki G, Kumamaru K, Kagawa H, Nagaya H, Sonh YC (2010): Production of recombinant Japanese eel gonadotropins by baculovirus in silkworm larvae. Gen Comp Endocrionol 167:379-386. https://doi.org/10.1016/j.ygcen.2010.01.003
  10. Lee SY, Byambaragchaa M, Kim JS, Seong HK, Kang MH, Min KS (2017): Biochemical characterization of recombination equine chorionic gonadotropin (receCG), using CHO cells and pathHunter parental cells expressing equine luteinizing hormone/chorionic gonadotropin receptors (eLH/CGR). J of Life Science 27:864-872.
  11. Min KS, Hiyama T, Seong HH, Hattori N, Tanaka S, Shiota K (2004): Biological activities of tethered equine chorionic gonadotropin (eCG) and its deglycosylated mutants. J Reprod Dev 50:297-304. https://doi.org/10.1262/jrd.50.297
  12. Moore WT, Ward DN (1980): Pregnant mare serum gonadotropin: an in vitro characterization of the luteotropin-follitropin dual activity. J Biol Chem 250:6930.
  13. Murphy BD, Martinuk SD (1991): Equine chorionic gonadotropin. Endocrine Reviews 12:27-44. https://doi.org/10.1210/edrv-12-1-27
  14. Ohta H, Miyake H, Miura C, Kamei H, Aida K, Miura T (2007): Follicle-stimulating hormone induces spermatogenesis mediated by androgen production in Japanese eel, Anguilla japonica. Biol Reprod 77:970-977. https://doi.org/10.1095/biolreprod.107.062299
  15. Ohta H, Sato Y, Imaizumi H, Kazeto Y (2017): Changes in milt volume and sperm quality with time after an injection of recombinant Japanese eel luteinizing hormone in male Japanese eels. Aquaculture 479:150-154. https://doi.org/10.1016/j.aquaculture.2017.05.044
  16. Park JJ, JarGal N, Yoon JT, Min KS (2009): Function of the tethered rec-eCG in rat and equine receptors. Reprod Dev Biol 33:229-236.
  17. Park JJ, JarGal N, Yoon JT, Min KS (2010): $\beta$-Subunit 94-96 residues of tethered recombinant equine chorionic gonadogropin are important sites luteinizing hormone and follicle stimulating hormone like activities. Reprod Dev Biol 34:33-40.
  18. Park JJ, Seong HK, Kim JS, Byambaragchaa M, Kang MH, Min KS (2017): Internalization of rat FSH and LH/CGR by rec-eCG in CHO-K1 cells. Dev Reprod 21:111-120. https://doi.org/10.12717/DR.2017.21.2.111
  19. Swanson P, Dickey JT, Campbell B (2003): Biochemistry and physiology of fish gonadotropins. Fish Physio Biochem 28:53-59. https://doi.org/10.1023/B:FISH.0000030476.73360.07