Fish Physiology and Biochemistry

, Volume 41, Issue 3, pp 587–601 | Cite as

Molecular cloning and characterization of gonadotropin subunits (GTHα, FSHβ and LHβ) and their regulation by hCG and GnRHa in Japanese sea bass (Lateolabrax japonicas) in vivo

  • Mei L. Chi
  • Meng Ni
  • Ji F. Li
  • Feng He
  • Kun Qian
  • Pei Zhang
  • Sen H. Chai
  • Hai S. Wen


In this study, three cDNA sequences encoding common glycoprotein α subunit (GTHα), follicle-stimulating hormone β subunit (FSHβ) and luteinizing hormone β subunit (LHβ) were isolated from Japanese sea bass (Lateolabrax japonicas). Comparison of the deduced amino acid sequences with other gonadotropic hormones (GTHs) indicated that their cysteine residues and potential N-linked glycosylation sites were highly conserved, and high homology with those of other perciformes was showed in phylogenetic analysis. GTHs transcripts were present highly in the pituitary and brain and weakly in testis and other tissues. During testicular development, GTHs transcriptional levels in pituitary and brain (expect FSHβ subunit in brain) were significantly increased at spermiation period, stage V. Subsequently, the effects of hCG and GnRHa on the mRNA levels of GTHs subunits were examined. In brain, both hormones were detected to improve the expression of GTHα subunit mRNA. In pituitary, three GTHs subunits increased parallelly and abruptly in two hormone treatment groups. In testis, hCG was suggested to improve three GTHs subunits expression in Japanese sea bass for the first time. These results suggest that both gonadotropins are probably involved in the control of Japanese sea bass spermatogenesis and provide a framework for better understanding of the mechanisms of hormone-mediated reproduction control in Japanese sea bass and other teleosts.


Japanese sea bass Gonadotropin subunits Testis hCG and GnRHa administration 



Gonadotropic hormone


Follicle-stimulating hormone


Luteinizing hormone


Human chorionic gonadotropin


Gonadotropin-releasing hormone analogue


Moloney murine leukemia virus


Physiological saline


Primordial germ cell


Pregnant mare serum gonadotropin


Coding sequence


National Center for Biotechnical Information



This research was supported by the National Key Technologies R&D Program (2011BAD13B03).


  1. Aizen J, Meiri I, Tzchori I, Levavi-Sivan B, Rosenfeld H (2005) Enhancing spawning in the grey mullet (Mugil cephalus) by removal of dopaminergic inhibition. Gen Comp Endocrinol 142:212–221CrossRefPubMedGoogle Scholar
  2. Ando H, Urano A (2005) Molecular regulation of gonadotropin secretion by gonadotropin-releasing hormone in salmonid fishes. Zool Sci 22:379–389CrossRefPubMedGoogle Scholar
  3. Apaja PM, Harju KT, Aatsinki JT, Petaja-Repo UE, Rajaniemi HJ (2004) Identification and structural characterization of the neuronal luteinizing hormone receptor associated with sensory systems. J Biol Chem 279:1899–1906CrossRefPubMedGoogle Scholar
  4. Bendtsen JD, Nielsen H, von Heijne G, Brunak S (2004) Improved prediction of signal peptides: SignalP 3.0. J Mol Biol 340:783–795CrossRefPubMedGoogle Scholar
  5. Biran J, Ben-Dor S, Levavi-Sivan B (2008) Molecular identification and functional characterization of the kisspeptin/kisspeptin receptor system in lower vertebrates. Biol Reprod 79:776–786CrossRefPubMedGoogle Scholar
  6. Breton B, Govoroun M, Mikolajczyk T (1998) GTHI and GTHII secretion profiles during the reproductive cycle in female rainbow trout: relationship with pituitary responsiveness to GnRHa stimulation. Gen Comp Endocrinol 111:38–50CrossRefPubMedGoogle Scholar
  7. Choi CY, An KW, Jo PG, Kang DY, Chang YJ (2007) Effects of gonadotropin-releasing hormone analog (GnRHa) on steroidogenic factor-1 (SF-1) and estrogen receptor β (ERβ) gene expression in the black porgy (Acanthopagrus schlegeli). Comp Biochem Phys B 147:82–86CrossRefGoogle Scholar
  8. Cohn WB, Jones RA, Valverde RA, Leiner KA, MacKenzie DS (2010) Molecular cloning and regulation of mRNA expression of the thyrotropin β and glycoprotein hormone α subunits in red drum, Sciaenops ocellatus. Fish Physiol Biochem 36:1277–1290CrossRefPubMedGoogle Scholar
  9. Dabrowski K, Ciereszko A, Ramseyer L, Culver D, Kestemont P (1994) Effects of hormonal treatment on induced spermiation and ovulation in the yellow perch Perca flavescens. Aquaculture 120:171–180CrossRefGoogle Scholar
  10. Dickey JT, Swanson P (2000) Effects of salmon gonadotropin-releasing hormone on follicle stimulating hormone secretion and subunit gene expression in coho salmon (Oncorhynchus kisutch). Gen Comp Endocrinol 118:436–449CrossRefPubMedGoogle Scholar
  11. Elisio M, Soria FN, Fernandino JI, Strüssmann CA, Somoza GM, Miranda LA (2012) Extrahypophyseal expression of gonadotropin subunits in pejerrey Odontesthes bonariensis and effects of high water temperatures on their expression. Gen Comp Endocrinol 175:329–336CrossRefPubMedGoogle Scholar
  12. Emanuele N, Anderson J, Baker G (1984) Release of immunoreactive luteinizing hormone (LH) from rat hypothalamus–pars tuberalis explants. Brain Res 309:271–276CrossRefPubMedGoogle Scholar
  13. Gen K, Okuzawa K, Senthilkumaran B, Tanaka H, Moriyama S, Kagawa H (2000) Unique expression of gonadotropin-I and II subunit genes in male and female red seabream (Pagrus major) during sexual maturation. Biol Reprod 6:308–319CrossRefGoogle Scholar
  14. Gomez JM, Weil C, Ollitrault M, Le Bail PY, Breton B, Le Gac F (1999) Growth hormone (GH) and gonadotropin subunit gene expression and pituitary and plasma changes during spermatogenesis and oogenesis in rainbow trout (Oncorhynchus mykiss). Gen Comp Endocrinol 113:413–428CrossRefPubMedGoogle Scholar
  15. Guzmán JM, Rubio M, Ortiz-Delgado JB, Klenke U, Kight K, Cross I, Sánchez-Ramos I, Riaza A, Rebordinos L, Sarasquete C, Zohar Y, Mañanós EL (2009a) Comparative gene expression of gonadotropins (FSH and LH) and peptide levels of gonadotropin-releasing hormones (GnRHs) in the pituitary of wild and cultured Senegalese sole (Solea senegalensis) broodstocks. Comp Biochem Physiol A 153:266–277CrossRefGoogle Scholar
  16. Guzmán JM, Bayarri MJ, Ramos J, Zohar Y, Sarasquete C, Mañanós EL (2009b) Follicle stimulating hormone (FSH) and luteinizing hormone (LH) gene expression during larval development in Senegalese sole (Solea senegalensis). Comp Biochem Physiol A 154:37–43CrossRefGoogle Scholar
  17. Hassin S, Gothilf Y, Blaise O, Zohar Y (1998) Gonadotropin-I and -II subunit gene expression of male striped bass (Morone saxatilis) after gonadotropin-releasing hormone analogue injection: quantitation using an optimized ribonuclease protection assay. Biol Reprod 58:1233–1240CrossRefPubMedGoogle Scholar
  18. Hassin S, Holland MCH, Zohar Y (2000) Early maturity in the male striped bass, Morone saxatilis: follicle-stimulating hormone and luteinizing hormone gene expression and their regulation by gonadotropin-releasing hormone analogue and testosterone. Biol Reprod 63:1691–1697CrossRefPubMedGoogle Scholar
  19. Hostetter G, Gallo RV, Brownfield MS (1987) Presence of immunoreactive luteinizing hormone in the rat forebrain. Neuroendocrinology 33:241–245CrossRefGoogle Scholar
  20. Huang H, Zhang Y, Huang WR, Li SS, Zhu P, Liu Y, Yin SW, Liu XC, Lin HR (2009) Molecular characterization of marbled eel (Anguilla marmorata) gonadotropin subunits and their mRNA expression profiles during artificially induced gonadal development. Gen Comp Endocrinol 162:192–202CrossRefPubMedGoogle Scholar
  21. Julenius K, Mølgaard A, Gupta R, Brunak S (2005) Prediction, conservation analysis and structural characterization of mammalian mucin-type O-glycosylation sites. Glycobiology 15:153–164CrossRefPubMedGoogle Scholar
  22. Kajimura S, Yoshiura Y, Suzuki M, Aida K (2001) cDNA cloning of two gonadotropin subunits (GTH-Iβ and -IIβ) and their expression profiles during gametogenesis in the Japanese flounder (Paralichthys olivaceus). Gen Comp Endocrinol 122:117–129CrossRefPubMedGoogle Scholar
  23. Kamei H, Ohira T, Yoshiura Y, Uchida N, Nagasawa H, Aida K (2003) Expression of a biologically active recombinant follicle stimulating hormone of Japanese eel Anguilla japonica using methylotropic yeast, Pichia pastoris. Gen Comp Endocrinol 134:244–254CrossRefPubMedGoogle Scholar
  24. Khan IA, Hawkins MB, Thomas P (1999) Gonadal stage-dependent effects of gonadal steroids on gonadotropin II secretion in the Atlantic croaker (Micropogonias undulatus). Biol Reprod 61:834–841CrossRefPubMedGoogle Scholar
  25. Kitahashi T, Ogawa S, Parhar IS (2009) Cloning and expression of kiss2 in the zebrafish and medaka. Endocrinology 150:821–831CrossRefPubMedGoogle Scholar
  26. Klausen C, Chang JP, Habibi HR (2002) Time- and dose-related effects of gonadotropin-releasing hormone on growth hormone and gonadotropin subunit gene expression in the goldfish pituitary. Can J Physiol Pharmacol 80:915–924CrossRefPubMedGoogle Scholar
  27. Klenke U, Zohar Y (2003) Gonadal regulation of gonadotropin subunit expression and pituitary LH protein content in female hybrid striped bass. Fish Physiol Biochem 28:25–27CrossRefGoogle Scholar
  28. Ko H, Park W, Kim DJ, Kobayashi M, Sohn YC (2007) Biological activities of recombinant Manchurian trout FSH and LH: their receptor specificity, steroidogenic and vitellogenic potencies. J Mol Endocrinol 38:99–111CrossRefPubMedGoogle Scholar
  29. Kobayashi M, Morita T, Ikeguchi K, Yoshizaki G, Suzuki T, Watabe S (2006) In vivo biological activity of recombinant goldfish gonadotropins produced by baculovirus in silkworm larvae. Aquaculture 256:433–442CrossRefGoogle Scholar
  30. Larsen DA, Swanson P (1997) Effects of gonadectomy on plasma gonadotropins I and II in Coho Salmon, Oncorhynchus kisutch. Gen Comp Endocrinol 108:152–160CrossRefPubMedGoogle Scholar
  31. Levavi-Sivan B, Bogerd J, Mañanós EL, Gómez A, Lareyre JJ (2010) Perspectives on fish gonadotropins and their receptors. Gen Comp Endocrinol 165:412–437CrossRefPubMedGoogle Scholar
  32. Li CJ, Zhou L, Wang Y, Hong YH, Gui JF (2005) Molecular and expression characterization of three gonadotropin subunits common α, FSH β and LHβ in groupers. Mol Cell Endocrinol 233:33–46CrossRefPubMedGoogle Scholar
  33. Mateos J, Mananos E, Martinez-Rodriguez G, Carrillo M, Querat B, Zanuy S (2003) Molecular characterization of sea bass gonadotropin subunits (α, FSHβ, and LHβ) and their expression during the reproductive cycle. Gen Comp Endocrinol 133:216–232CrossRefPubMedGoogle Scholar
  34. Mayes KB, Rosenblum PM, Brandt TM (1993) Raceway spawning of Florida largemouth bass: effects of acclimation time and hormone treatment on spawning success. Prog Fish-Cult 55:1–8CrossRefGoogle Scholar
  35. McCann SM, Kimura M, Walczewska A, Karanth S, Rettori V, Yu WH (1998) Hypothalamic control of FSH and LH by FSH-RF, LHRH, cytokines, leptin and nitric oxide. Neuroimmunomodulation 5:193–202CrossRefPubMedGoogle Scholar
  36. Mechaly AS, Viñas J, Piferrer F (2012) Sex-specific changes in the expression of kisspeptin, kisspeptin receptor, gonadotropins and gonadotropin receptors in the Senegalese sole (Solea senegalensis) during a full reproductive cycle. Comp Biochem Physiol A 162:364–371CrossRefGoogle Scholar
  37. Muralidhar K, Moudgal NR (1976) Studies on rat ovarian receptors for lutropin (luteinizing hormone). Interaction with β-subunit of sheep lutropin. Biochem J 160:615–619PubMedCentralPubMedGoogle Scholar
  38. Nagae M, Todo T, Gen K, Kato Y, Young G, Adachi S, Yamauchi K (1996) Molecular cloning of the cDNAs encoding pituitary glycoprotein hormone and gonadotropin IIβ-subunits of the Japanese eel, Anguilla japonica, and increase in their mRNAs during ovarian development induced by injection of chum salmon pituitary homogenate. J Mol Endocrinol 16:171–181CrossRefPubMedGoogle Scholar
  39. Pandolfi M, Pozzi AG, Cánepa M, Vissio PG, Shimizu A, Maggese MC, Lobo G (2009) Presence of β-follicle-stimulating hormone and β-luteinizing hormone transcripts in the brain of Cichlasoma dimerus (Perciformes: Cichlidae), effect of brain-derived gonadotropins on pituitary hormone release. Neuroendocrinology 89:27–37CrossRefPubMedGoogle Scholar
  40. Parhar I, Soga T, Ogawa S, Sakuma Y (2003) FSH and LH-β subunits in the preoptic nucleus: ontogenic expression in teleost. Gen Comp Endocrinol 132:369–378CrossRefPubMedGoogle Scholar
  41. Pierce JG, Parsons TF (1981) Glycoprotein hormones: structure and function. Annu Rev Biochem 50:465–495CrossRefPubMedGoogle Scholar
  42. Prat F, Sumpter JP, Tyler CR (1996) Validation of radioimmunoassays 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 54:1375–1382CrossRefPubMedGoogle Scholar
  43. Quérat B, Sellouk A, Salmon C (2000) Phylogenetic analysis of the vertebrate glycoprotein hormone family including new sequences of sturgeon (Acipenser baeri) β subunits of the two gonadotropins and the thyroid-stimulating hormone. Biol Reprod 63:222–228CrossRefPubMedGoogle Scholar
  44. Schirman-Hildesheim TD, Gershon E, Litichever N, Galiani D, Ben-Aroya N, Dekel N, Koch Y (2008) Local production of the gonadotropic hormones in the rat ovary. Mol Cell Endocrinol 282:32–38CrossRefPubMedGoogle Scholar
  45. Schulz RW, Vischer HF, Cavaco JE, Santos EM, Tyler CR, Goos HJ, Bogerd J (2001) Gonadotropins, their receptors, and the regulation of testicular functions in fish. Comp Biochem Physiol B 129:407–417CrossRefPubMedGoogle Scholar
  46. So WK, Kwok HF, Ge W (2005) Zebrafish gonadotropins and their receptors: II. Cloning and characterization of zebrafish follicle-stimulating hormone and luteinizing hormone subunits–their spatial-temporal expression patterns and receptor specificity. Biol Reprod 72:1382–1396CrossRefPubMedGoogle Scholar
  47. Swanson P, Dickey JT, Campbell B (2003) Biochemistry and physiology of fish gonadotropins. Fish Physiol Biochem 28:53–59CrossRefGoogle Scholar
  48. Ulloa-Aguirre A, Timossi C, Damian-Matsumura P, Dias JA (1999) Role of glycosylation in function of follicle-stimulating hormone. Endocrine 11:205–215CrossRefPubMedGoogle Scholar
  49. Vischer HF, Granneman JCM, Linskens MHK, Schulz RW, Bogerd J (2003) Both recombinant African catfish LH and FSH are able to activate the African catfish FSH receptor. J Mol Endocrinol 31:133–140CrossRefPubMedGoogle Scholar
  50. Watanabe WO, Ellis EP, Ellis SC, Chaves J, Manfredi C, Hagood RW, Sparsis M, Arneson S (1998) Artificial propagation of mutton snapper Lutjanus analis, a new candidate marine fish species for aquaculture. J World Aquacult Soc 29:176–187CrossRefGoogle Scholar
  51. Weltzien FA, Kobayashi T, Andersson E, Norberg B, Andersen O (2003) Molecular characterization and expression of FSH beta, LH beta, and common alpha-subunit in male Atlantic halibut (Hippoglossus hippoglossus). Gen Comp Endocrinol 131:87–96CrossRefPubMedGoogle Scholar
  52. Wong TT, Zohar Y (2004) Novel expression of gonadotropin subunit genes in oocytes of the gilthead seabream (Sparus aurata). Endocrinology 145:5210–5220CrossRefPubMedGoogle Scholar
  53. Wu F, Zhang X, Zhang W, Huang B, Liu Z, Hu C, Wang D (2009) Expression of three gonadotropin subunits in Southern catfish gonad and their possible roles during early gonadal development. Comp Biochem Physiol A 153:44–48CrossRefGoogle Scholar
  54. Yaron Z, Gur G, Melamed P, Rosenfeld H, Levavi-Sivan B, Elizur A (2001) Regulation of gonadotropin subunit genes in tilapia. Comp Biochem Physiol B 129:489–502CrossRefPubMedGoogle Scholar
  55. Yaron Z, Gur G, Melamed P, Rosenfeld H, Elizur A, Levavi-Sivan B (2003) Regulation of fish gonadotropins. Int Rev Cytol 225:131–185CrossRefPubMedGoogle Scholar
  56. Yuen CW, Ge W (2004) Follistatin suppresses FSHβ but increases LHβ expression in the goldfish—evidence for an activin-mediated autocrine/paracrine system in fish pituitary. Gen Comp Endocrinol 135:108–115CrossRefPubMedGoogle Scholar
  57. Zenkevics H, Vose V, Vosekalne I, Bũcena A (2000) Partial deglycosylation of alpha subunit modifies sturgeon gonadotropin function. Acta Biochim Pol 47:815–819PubMedGoogle Scholar
  58. Zhang CD, Li MY (2005) Advance of study on reproductive biology and breeding technology of Lateolabrax japonicus. J Ningbo Univ 18:400–403 (Chinese with English abstract)Google Scholar
  59. Zhang MZ, Gao TX, Ruan SH, Zhuang QZ, Qu YQ (2001) Study on artificial cultivation and spawning inducement technique of Lateolabrax japonicus. Ocean Univ Qingdao 31:195–200 (Chinese with English abstract)Google Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Mei L. Chi
    • 1
  • Meng Ni
    • 1
  • Ji F. Li
    • 1
  • Feng He
    • 1
  • Kun Qian
    • 1
  • Pei Zhang
    • 1
  • Sen H. Chai
    • 1
  • Hai S. Wen
    • 1
  1. 1.Fisheries CollegeOcean University of ChinaQingdaoChina

Personalised recommendations