Korean Journal of Fisheries and Aquatic Sciences (한국수산과학회지)

The Korean Society of Fisheries and Aquatic Science (한국수산과학회)
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Isolation and Identification of FSHβ and LHβ Subunits in the Blacktip Grouper Epinephelus fasciatus

홍바리(Epinephelus fasciatus) FSHβ와 LHβ subunit의 분리 및 동정

  • Park, Yong-Ju (Marine and Environmental Research Institute, Jeju National University) ;
  • Kang, Hyeong-Cheol (Marine and Environmental Research Institute, Jeju National University) ;
  • Lee, Chi-Hoon (Marine and Environmental Research Institute, Jeju National University) ;
  • Song, Young-Bo (Department of Biology, Jeju National University) ;
  • Baek, Hea-Ja (Department of Marine Biology, Pukyong National University) ;
  • Kim, Hyung-Bae (Department of Marine Bio-Resources, Gangwon Provincial University) ;
  • Soyano, Kiyoshi (Institute for East China Sea Research, Nagasaki University) ;
  • Lee, Young-Don (Marine and Environmental Research Institute, Jeju National University)
  • 박용주 (제주대학교 해양과환경연구소) ;
  • 강형철 (제주대학교 해양과환경연구소) ;
  • 이치훈 (제주대학교 해양과환경연구소) ;
  • 송영보 (제주대학교 생물학과) ;
  • 백혜자 (부경대학교 자원생물학과) ;
  • 김형배 (강원도립대학 해양생명과학과) ;
  • ;
  • 이영돈 (제주대학교 해양과환경연구소)
  • Received : 2011.05.12
  • Accepted : 2012.02.02
  • Published : 2012.02.29
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Abstract

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.

Keywords

References

  1. Ando H, Swanson P, Kitani T, Koide N, Okada H, Ueda H and Urano A. 2004. Synergistic effects of salmon gonadotropin- releasing hormone and estradiol-${17}\beta$ on gonadotropin subunit gene expression and release in masu pituitary cells in vitro. Gen Comp Endocrinol 137, 109-121. https://doi.org/10.1016/j.ygcen.2004.02.012
  2. Banerjee A and Khan IA. 2008. Molecular cloning of FSH and ${LH}\beta$ subunit and their regulation by estrogen in Atlantic croaker. Gen Comp Endocrinol 155, 827-837. https://doi.org/10.1016/j.ygcen.2007.09.016
  3. Dyrlov BJ, Nielsen H, Heijne VG and Brunak S. 2004. Improved prediction of signal peptides: Signal P 3.0. J Mol Biol 340, 783-795. https://doi.org/10.1016/j.jmb.2004.05.028
  4. Hellqvist A, Bornestaf C, Borg B and Schmitz M. 2004. Cloning and sequencing of the FSH$\beta$- and LH$\beta$-subunit in the three-spined stickleback, Gasterosteus aculeatus, and effects of photoperiod and temperature on LH$\beta$ and FSH$\beta$ mRNA expression. Gen Comp Endocrinol 135, 167-174. https://doi.org/10.1016/j.ygcen.2003.09.003
  5. Hostetter G, Gallo RV and Brownfield MS. 1981. Presence of immunoreactive luteining hormone in the rat forebrain. Neuroendocrinology 33, 241-245. https://doi.org/10.1159/000123238
  6. Huggard-Nelson DL, Nathwani PS, Kermouni A and Habibi HR. 2002. Molecular characterization of LH-beta and FSHbeta subunits and their regulation by estrogen in the goldfish pituitary. Mol Cell Endocrinol 188, 171-193. https://doi.org/10.1016/S0303-7207(01)00716-X
  7. Jackson K, Goldberg D, Ofir M, Abraham M and Degani G. 1999. Blue gourami (Trichogaster trichopterus) gonadotropic beta subunits (I and II) cDNA sequences and expression during oogenesis. J Mol Endocrinol 23, 177-187. https://doi.org/10.1677/jme.0.0230177
  8. Kajimura S, Yoshiura Y, Suzuki M and Aida K. 2001. cDNA cloning of two gonadotropin beta subunits (GTH-I beta and-II beta) and their expression profiles during gametogenesis in the Japanese flounder (Paralichthys olivaceus). Gen Comp Endocrinol 122, 117-129. https://doi.org/10.1006/gcen.2000.7610
  9. Ko H, Park W, Kim DJ, Kobayashi M and Sohn YC. 2007. Biological activities of recombinant Manchurian trout FSH and LH: their receptor specificity, steroidogenic and vitellogenic potencies. J Mol Endocrinol 38, 99-111. https://doi.org/10.1677/jme.1.02163
  10. Kobayashi MA, Sohn YC, Yoshiura YA and Aida KA. 2000. Effects of sex steroids on the mRNA levels of gonadotropin subunits in juvenile and ovariectomized goldfish Carassius auratus. Fish Sci 66, 223-231. https://doi.org/10.1046/j.1444-2906.2000.00038.x
  11. Kobayashi M, Morita T, Ikeguchi K, Yoshizaki G, Suzuki T and Watabe S. 2006. In vivo biological activity of recombinant goldfish gonadotropins produced by baculovirus in silkworm larvae. Aquaculture 256, 433-442. https://doi.org/10.1016/j.aquaculture.2006.01.016
  12. Lin YW, Rupnow BA, Price DA, Greenberg RM and Wallace RA. 1992. Fundulus heteroclitus gonadotropins. 3. Cloning and sequencing of gonadotropic hormone (GTH) I and II$\beta$ subunits using the polymerase chain reaction. Mol Cell Endocrinol 85, 127-139. https://doi.org/10.1016/0303-7207(92)90132-P
  13. Martyniuk CJ, Kroll KJ, Porak WF, Steward C, Grier HJ and Denslow ND. 2009. Seasonal relationship between gonadotropin, growth hormone, and estrogen receptor mRNA expression in the pituitary gland of largemouth bass. Gen Comp Endocrinol 163, 306-317. https://doi.org/10.1016/j.ygcen.2009.04.028
  14. Mateos J, Mananos E, Carrillo M and Zanuy S. 2002. Regulation of follicle-stimulating hormone (FSH) and luteinizing hormone (LH) gene expression by gonadotropinreleasing hormone (GnRH) and sexual steroids in the Mediterranean Sea bass. Comp Biochem Physiol 132B, 75-86.
  15. Melamed P, Rosenfeld H, Elizur A and Yaron Z. 1998. Endocrine regulation of gonadotropin and growth hormone gene transcription in fish. Comp Biochem Physiol 119C, 325-338.
  16. Nielsen H, Engelbrecht J, Brunak S and Heijne VG. 1997. Identification of prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites. Protein Eng 10, 1-6. https://doi.org/10.1093/protein/10.1.1
  17. Nozaki M, Naito N, Swanson P, Dickhoff WW, Nakai Y, Suzuki K and Kawauchi H. 1990a. Salmonid pituitary gonadotrophs. II. Ontogeny of GTH I and GTH II cells in the rainbow trout (Salmo gairdneriirideus). Gen Comp Endocrinol 77, 358-367. https://doi.org/10.1016/0016-6480(90)90225-B
  18. Nozaki M, Naito N, Swanson P, Miyata K, Nakai Y, Oota Y, Suzuki K and Kawauchi H. Salmonid pituitary gonadotrophs. I. Distinct cellular distributions of two gonadotropins, GTH I and GTH II. Gen Comp Endocrinol 77, 348-357. https://doi.org/10.1016/0016-6480(90)90224-A
  19. Parhar IS, Soga T, Ogawa S and Sakuma Y. 2003. FSH and LH-$\beta$ subunits in the preoptic nucleus: ontogenic expression in teleost. Gen Comp Endocrinol 132, 369-378. https://doi.org/10.1016/S0016-6480(03)00121-7
  20. Pierce JG and Parsons TF. 1981. Glycoprotein hormones: structure and function. Annu Rev Biochem 50, 464-495.
  21. Prat F, Sumpter JP and Tyler CR. 1996. Validation of radioimmunoassay for two salmon gonadotropins (GTH I and GTH II) and their plasma concentrations throughout the reproductive cycle in male and female rainbow trout (Oncorhynchus mykiss). Biol Reprod 54, 1375-1382. https://doi.org/10.1095/biolreprod54.6.1375
  22. Randall JE and Heemstra PC. 1991. Epinephelus fasciatus. Revision of Indo-Pacific groupers (Perciformes: Serranidae: Epinephelinae), with descriptions of five new species. Indo-Pacific fishes No. 20. Bernice Pauahi Bishop Museum, Honolulu.
  23. Schally AV, Arimura A, Kastin AJ, Matsuo H, Baba Y, Redding TW, Nair RM, Debeljuk L and White WF. 1971. Gonadotropin-releasing hormone: One polypeptide regulates secretion of luteinizing and follicle-stimulating hormones. Science 173, 1036-1038. https://doi.org/10.1126/science.173.4001.1036
  24. Sekine S, Saito A, Itoh H, Kawauchi H and Itoh S. 1989. Molecular cloning and sequence analysis of chum salmon gonadotropin cDNAs. Proc Natl Acad Sci USA 86, 8645- 8649. https://doi.org/10.1073/pnas.86.22.8645
  25. Sohn YC, Kobayashi M and Aida K. 2001. Regulation of gonadotropin b subunit gene expression by testosterone and gonadotropin-releasing hormones in the goldfish, Carassius auratus. Comp Biochem Physiol B 129, 419-426. https://doi.org/10.1016/S1096-4959(01)00342-6
  26. Sower SA, Moriyama S, Kasahara M, Takahashi A, Nozaki M, Uchida K, Dahlstrom JM and Kawauchi H. 2006. Identification of sea lamprey GTHb-like cDNA and its evolutionary implications. Gen Comp Endocrinol 148, 22-32. https://doi.org/10.1016/j.ygcen.2005.11.009
  27. Swanson P, Suzuki K, Kawauchi H and Dickhoff WW. 1991. Isolation and characterization of two coho salmon gonadotropins, GTH I and GTH II. Biol Reprod 44, 29-38. https://doi.org/10.1095/biolreprod44.1.29
  28. Trinh KY, Wang NC, Hew CL and Crim LW. 1986. Molecular cloning and sequencing of salmon gonadotropin $\beta$ subunit. Eur J Biochem 159, 619-624. https://doi.org/10.1111/j.1432-1033.1986.tb09930.x
  29. Trofimova IN and Belikov SI. 1994. Cloning and sequencing the cDNA for the $\beta$ subunit of Baikal omul gonadotropin. Mol Biol (Moscow) 28, 1052-1056.
  30. Yaron Z, Gur G, Melamed P, Rosenfeld H, Levavi-Sivan B and Elizur A. 2001. Regulation of gonadotropin subunit genes in tilapia. Comp Biochem Physiol 129B, 489-502.
  31. Yu JYL, Namiki H and Gorbman A. 1979. Rat gonadotropin release stimulated in vitro by GnRH-depleted rat brain extracts. Neuroendocrinology 29, 54-65. https://doi.org/10.1159/000122905

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