Abstract
The sapphire devil (Chrysiptera cyanea) is a tropical damselfish that undergoes active reproduction under long-day conditions. To elucidate the physiological regulation of the brain–pituitary–gonadal axis in female sapphire devil, we cloned and characterized the genes of two kisspeptins (kiss1 and kiss2), three gonadotropin-releasing hormones (gnrh1, gnrh2, gnrh3), and the β-subunit of two gonadotropins (fshβ and lhβ) and investigated the gene expression changes during ovarian development. Quantitative polymerase chain reaction analyses in various brain parts revealed high expression levels of kiss1, kiss2, and gnrh2 in the diencephalon; gnrh2 and gnrh3 in the telencephalon; and fshβ and lhβ in the pituitary. In situ hybridization (ISH) analyses revealed positive signals of kiss1 in the dorsal and ventral habenular nucleus and of kiss2 in the dorsal and ventral parts of the nucleus of the lateral recess. This analysis showed gnrh1 expression in the preoptic area (POA), suggesting that GnRH1 plays a stimulating role in the secretion of gonadotropins from the pituitary of the sapphire devil. High transcription levels of kiss1, kiss2, gnrh1, gnrh2, fshβ, and lhβ were observed in the brain during the late vitellogenic stage, suggesting their involvement in the physiological processes of vitellogenesis. Immersion of fish in estradiol-17β (E2)-containing seawater resulted in increased expression of kiss2 and gnrh1 in their brains. This study showed that kiss-expressing neurons in the diencephalon are influenced by E2, leading to upregulation of gnrh1 in the POA and of fshβ and lhβ in the pituitary during vitellogenesis.
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References
Alvarado MV, Carrillo M, Felip A (2013) Expression of kisspeptins and their receptors, gnrh-1/gnrhr-II-1a and gonadotropin genes in the brain of adult male and female European sea bass during different gonadal stages. Gen Comp Endocrinol 187:104–116. https://doi.org/10.1016/j.ygcen.2013.03.030
Amano M, Pham KX, Amiya N et al (2008) Changes in brain seabream GnRH mRNA and pituitary seabream GnRH peptide levels during ovarian maturation in female barfin flounder. Gen Comp Endocrinol 158:168–172. https://doi.org/10.1016/j.ygcen.2008.06.008
Badruzzaman M, Bapary MAJ, Takemura A (2013) Possible roles of photoperiod and melatonin in reproductive activity via changes in dopaminergic activity in the brain of a tropical damselfish, Chrysiptera cyanea. Gen Comp Endocrinol 194:240–247. https://doi.org/10.1016/j.ygcen.2013.09.012
Badruzzaman M, Imamura S, Takeuchi Y, Ikegami T, Takemura A (2015) Effects of neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) treatment on ovarian development of the sapphire devil, Chrysiptera cyanea. Fish Physiol Biochem 41:61–71. https://doi.org/10.1007/s10695-014-0006-9
Bapary MAJ, Takemura A (2010) Effect of temperature and photoperiod on the reproductive condition and performance of a tropical damselfish Chrysiptera cyanea during different phases of the reproductive season. Fisheries Sci 76:769–776. https://doi.org/10.1007/s12562-010-0272-0
Bapary MAJ, Fainuulelei P, Takemura A (2009) Environmental control of gonadal development in the tropical damselfish Chrysiptera cyanea. Mar Biol Res 5:462–469. https://doi.org/10.1080/17451000802644722
Bapary MAJ, Amin MN, Takemura A (2012) Food availability as a possible determinant for initiation and termination of reproductive activity in the tropical damselfish Chrysiptera cyanea. Mar Biol Res 8:154–162. https://doi.org/10.1080/17451000.2011.605146
Bayarri MJ, Rodríguez L, Zanuy S et al (2004) Effect of photoperiod manipulation on the daily rhythms of melatonin and reproductive hormones in caged European sea bass (Dicentrarchus labrax). Gen Comp Endocrinol 136:72–81. https://doi.org/10.1016/j.ygcen.2003.12.004
Burns RS, Chiueh CC, Markey SP et al (1983) A primate model of parkinsonism: selective destruction of dopaminergic neurons in the pars compacta of the substantia nigra by N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine. P Natl Acad Sci USA 80:4546–4550
Cerdá J, Chauvigne F, Agulleiro MJ et al (2008) Molecular cloning of Senegalese sole (Solea senegalensis) follicle-stimulating hormone and luteinizing hormone subunits and expression pattern during spermatogenesis. Gen Comp Endocrinol 156:470–481. https://doi.org/10.1016/j.ygcen.2008.02.006
Chai K, Liu X, Zhang Y, Lin H (2013) Day-night and reproductive cycle profiles of melatonin receptor, kiss, and gnrh expression in orange-spotted grouper (Epinephelus coioides). Mol Reprod Dev 80:535–548. https://doi.org/10.1002/mrd.22191
Escobar S, Felip A, Gueguen MM et al (2013) Expression of kisspeptins in the brain and pituitary of the european sea bass (Dicentrarchus labrax). J Comp Neurol 521:933–948. https://doi.org/10.1002/cne.23211/pdf
Falcón J, Migaud H, Muñoz-Cueto J-A, Carrillo M (2010) Current knowledge on the melatonin system in teleost fish. Gen Comp Endocrinol 165:469–482. https://doi.org/10.1016/j.ygcen.2009.04.026
Felip A, Zanuy S, Pineda R et al (2009) Evidence for two distinct KiSS genes in non-placental vertebrates that encode kisspeptins with different gonadotropin-releasing activities in fish and mammals. Mol Cell Endocrinol 312:61–71. https://doi.org/10.1016/j.mce.2008.11.017
Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791. https://doi.org/10.2307/2408678
Filby AL, van Aerle R, Duitman J, Tyler CR (2008) The kisspeptin/gonadotropin-releasing hormone pathway and molecular signaling of puberty in fish. Biol Reprod 78:278–289. https://doi.org/10.1095/biolreprod.107.063420
Forniés MA, Carrillo M, Mañanós E et al (2003) Relative potency of the forms of GnRH and their analogs on LH release in sea bass. J Fish Biol 63:73–89. https://doi.org/10.1046/j.1095-8649.2003.00130.x
Geffard M, Kah O, Chambolle P et al (1982) 1st immunocytochemical application of an anti-dopamine antibody in the study of the central nervous system. C R Seances Acad Sci III 295:797–802
González-Martínez D, Madigou T, Zmora N et al (2001) Differential expression of three different prepro-GnRH (gonadotrophin-releasing hormone) messengers in the brain of the European sea bass (Dicentrarchus labrax). J Comp Neurol 429:144–155. https://doi.org/10.1002/1096-9861(20000101)429:1<144::AID-CNE11>3.0.CO;2-B
González-Martínez D, Zmora N, Mañanos E et al (2002) Immunohistochemical localization of three different prepro-GnRHs in the brain and pituitary of the European sea bass (Dicentrarchus labrax) using antibodies to the corresponding GnRH-associated peptides. J Comp Neurol 446:95–113. https://doi.org/10.1002/cne.10190
González-Martínez D, Zmora N, Saligaut D et al (2004) New insights in developmental origins of different GnRH (gonadotrophin-releasing hormone) systems in perciform fish: an immunohistochemical study in the European sea bass (Dicentrarchus labrax). J Chem Neuroanat 28:1–15. https://doi.org/10.1016/j.jchemneu.2004.05.001
Gothilf Y, Muñoz-Cueto J-A, Sagrillo CA et al (1996) Three forms of gonadotropin-releasing hormone in a perciform fish (Sparus aurata): complementary deoxyribonucleic acid characterization and brain localization. Biol Reprod 55:636–645. https://doi.org/10.1210/endo.141.2.7300
Heikkila R, Hess A, Duvoisin R (1984) Dopaminergic neurotoxicity of 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine in mice. Science 224:1451–1453. https://doi.org/10.1126/science.6610213
Herbison AE (1998) Multimodal influence of estrogen upon gonadotropin-releasing hormone neurons. Endocr Rev 19:302–330. https://doi.org/10.1210/edrv.19.3.0332
Hoque M, Takemura A, Takano K (1998) Annual changes in oocyte development and serum vitellogenin level in the rabbitfish Siganus canaliculatus (Park) in Okinawa, Southern Japan. Fisheries Sci 64:44–51. https://doi.org/10.1002/cne.903560105
Imamura S, Hur S-P, Takeuchi Y et al (2017) Molecular cloning of kisspeptin receptor genes (gpr54-1 and gpr54-2) and their expression profiles in the brain of a tropical damselfish during different gonadal stages. Comp Biochem Physiol A 203:9–16. https://doi.org/10.1016/j.cbpa.2016.07.015
Kah O, Chambolle P (1983) Serotonin in the brain of the goldfish, Carassius auratus. Cell Tissue Res 234:319–333. https://doi.org/10.1007/BF00213771
Kah O, Chambolle P, Olivereau M (1978) Aminergic innervation of the hypothalamo-hypophyseal system in Gambusia sp. (Teleost, Poecilidae) studied by two fluorescence techniques. C R Acad Sci Hebd Seances Acad Sci D 286:705–708
Kanda S, Akazome Y, Matsunaga T et al (2008) Identification of KiSS-1 product kisspeptin and steroid-sensitive sexually dimorphic kisspeptin neurons in medaka (Oryzias latipes). Endocrinology 149:2467–2476. https://doi.org/10.1210/en.2007-1503
Kanda S, Karigo T, Oka Y (2012) Steroid sensitive kiss2 neurones in the goldfish: evolutionary insights into the duplicate kisspeptin gene-expressing neurones. J Neuroendocrinol 24:897–906. https://doi.org/10.1111/j.1365-2826.2012.02296.x
Karigo T, Oka YP (2013) Neurobiological study of fish brains gives insights into the nature of gonadotropin-releasing hormone 1–3 neurons. Front Endocrinol. https://doi.org/10.3389/fendo.2013.00177
Karigo T, Kanda S, Takahashi A et al (2012) Time-of-day-dependent changes in GnRH1 neuronal activities and gonadotropin mRNA expression in a daily spawning fish, Medaka. Endocrinology 153:3394–3404. https://doi.org/10.1210/en.2011-2022
Kimura M (1991) The neutral theory of molecular evolution: a review of recent evidence. Jap Jo Geneti 66:367–386. https://doi.org/10.1266/jjg.66.367
Kitahashi T, Ogawa S, Parhar IS (2009) Cloning and expression of kiss2 in the zebrafish and medaka. Endocrinology 150:821–831. https://doi.org/10.1210/en.2008-0940
Larkin MA, Blackshields G, Brown NP et al (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948. https://doi.org/10.1093/bioinformatics/btm404
Lethimonier C, Madigou T, Muñoz-Cueto J-A et al (2004) Evolutionary aspects of GnRHs, GnRH neuronal systems and GnRH receptors in teleost fish. Gen Comp Endocrinol 135:1–16. https://doi.org/10.1016/j.ygcen.2003.10.007
Li S, Zhang Y, Liu Y et al (2009) Structural and functional multiplicity of the kisspeptin/GPR54 system in goldfish (Carassius auratus). J Endocrinol 201:407–418. https://doi.org/10.1677/JOE-09-0016
Mechaly AS, Viñas J, Piferrer F (2011) Gene structure analysis of kisspeptin-2 (Kiss2) in the Senegalese sole (Solea senegalensis): characterization of two splice variants of Kiss2, and novel evidence for metabolic regulation of kisspeptin signaling in non-mammalian species. Mol Cell Endocrinol 339:14–24. https://doi.org/10.1016/j.mce.2011.03.004
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 162:364–371. https://doi.org/10.1016/j.cbpa.2012.04.003
Mitani Y, Kanda S, Akazome Y et al (2013) Hypothalamic kiss1 but not kiss2 neurons are involved in estrogen feedback in medaka (Oryzias latipes). Endocrinology 151:1751–1759. https://doi.org/10.1210/en.2009-1174
Mohamed JS, Thomas P, Khan IA (2005) Isolation, cloning, and expression of three prepro-GnRH mRNAs in Atlantic croaker brain and pituitary. J Comp Neurol 488:384–395. https://doi.org/10.1002/cne.20596
Nagahama Y (1994) Endocrine regulation of gametogenesis in fish. Int J Dev Biol 38:217–229
Nakajo M, Kanda S, Karigo T et al (2018) Evolutionally conserved function of kisspeptin neuronal system is nonreproductive regulation as revealed by nonmammalian study. Endocrinology 159:163–183. https://doi.org/10.1210/en.2017-00808
Nocillado JN, Zohar Y, Biran J et al (2013) Chronic kisspeptin administration stimulated gonadal development in pre-pubertal male yellowtail kingfish (Seriola lalandi; Perciformes) during the breeding and non-breeding season. Gen Comp Endocrinol 191:168–176. https://doi.org/10.1016/j.ygcen.2013.06.005
Okubo K, Amano M, Yoshiura Y et al (2000) A novel form of gonadotropin-releasing hormone in the medaka, Oryzias latipes. Biochem Biophys Res Commun 276:298–303. https://doi.org/10.1006/bbrc.2000.3476
Pandolfi M, Cueto JAM, Nostro Lo FL et al (2005) GnRH systems of Cichlasoma dimerus (Perciformes, Cichlidae) revisited: a localization study with antibodies and riboprobes to GnRH-associated peptides. Cell Tissue Res 321:219–232. https://doi.org/10.1007/s00441-004-1055-7
Parhar IS, Ogawa S, Hamada T, Sakuma Y (2003) Single-cell real-time quantitative polymerase chain reaction of immunofluorescently identified neurons of gonadotropin-releasing hormone subtypes in cichlid fish. Endocrinology 144:3297–3300. https://doi.org/10.1210/en.2003-0386
Powell JFF, Zohar Y, Elizur A et al (1994) Three forms of gonadotropin-releasing hormone characterized from brains of one species. P Natl Acad Sci USA 91:12081–12085
Rahman MS, Takemura A, Takano K (2000) Annual changes in ovarian histology, plasma steroid hormones and vitellogenin in the female golden rabbitfish, Siganus guttatus (Bloch). Bull Mar Sci 67:729–740
Roa J, Aguilar E, Diéguez C et al (2008) New frontiers in kisspeptin/GPR54 physiology as fundamental gatekeepers of reproductive function. Front Neuroendocrinol 29:48–69. https://doi.org/10.1016/j.yfrne.2007.07.002
Selvaraj S, Selvaraj S, Kitano H et al (2010) Molecular characterization, tissue distribution, and mRNA expression profiles of two Kiss genes in the adult male and female chub mackerel (Scomber japonicus) during different gonadal stages. Gen Comp Endocrinol 169:28–38. https://doi.org/10.1016/j.ygcen.2010.07.011
Selvaraju S, Sivasubramani T, Raghavendra BS et al (2012) Effect of dietary energy on seminal plasma insulin-like growth factor-I (IGF-I), serum IGF-I and testosterone levels, semen quality and fertility in adult rams. Theriogenology 78:646–655. https://doi.org/10.1016/j.theriogenology.2012.03.010
Servili A, Le Page Y, Escobar S et al (2011) Organization of two independent kisspeptin systems derived from evolutionary-ancient kiss genes in the brain of zebrafish. Endocrinology 152:1527–1540. https://doi.org/10.1210/en.2010-0948
Shahjahan M, Hamabata T, Motohashi E et al (2010a) Differential expression of three types of gonadotropin-releasing hormone genes during the spawning season in grass puffer, Takifugu niphobles. Gen Comp Endocrinol 167:153–163. https://doi.org/10.1016/j.ygcen.2010.01.018
Shahjahan M, Motohashi E, Doi H, Ando H (2010b) Elevation of Kiss2 and its receptor gene expression in the brain and pituitary of grass puffer during the spawning season. Gen Comp Endocrinol 169:48–57. https://doi.org/10.1016/j.ygcen.2010.07.008
Smith JT, Cunningham MJ, Rissman EF et al (2005) Regulation of Kiss1 gene expression in the brain of the female mouse. Endocrinology 146:3686–3692. https://doi.org/10.1210/en.2005-0488
Smith JT, Clifton D, Steiner RA (2006) Regulation of the neuroendocrine reproductive axis by kisspeptin-GPR54 signaling. Reproduction 131:623–630. https://doi.org/10.1530/rep.1.00368
So W-K, Kwok H-F, 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 72:1382–1396. https://doi.org/10.1095/biolreprod.104.038216
Tang H, Liu Y, Luo D et al (2015) The kiss/kissr systems are dispensable for zebrafish reproduction: evidence from gene knockout studies. Endocrinology 156:589–599. https://doi.org/10.1210/en.2014-1204
Tena-Sempere M, Felip A, Gómez A et al (2012) Comparative insights of the kisspeptin/kisspeptin receptor system: lessons from non-mammalian vertebrates. Gen Comp Endocrinol 175:234–243. https://doi.org/10.1016/j.ygcen.2011.11.015
van Aerle R, Kille P, Lange A, Tyler CR (2008) Evidence for the existence of a functional Kiss1/Kiss1 receptor pathway in fish. Peptides 29:57–64. https://doi.org/10.1016/j.peptides.2007.10.018
Vickers ED, Laberge F, Adams BA et al (2004) Cloning and localization of three forms of gonadotropin-releasing hormone, including the novel whitefish form, in a salmonid, Coregonus clupeaformis. Biol Reprod 70:1136–1146. https://doi.org/10.1095/biolreprod.103.023846
Xue X, Ogawa S, Ng KW et al (2013) Thyroid hormone upregulates hypothalamic kiss2 gene in the male Nile tilapia, Oreochromis niloticus. Front Endocrinol 4:184. https://doi.org/10.3389/fendo.2013.00184
Yang B, Jiang Q, Chan T et al (2010) Goldfish kisspeptin: molecular cloning, tissue distribution of transcript expression, and stimulatory effects on prolactin, growth hormone and luteinizing hormone secretion and gene expression via direct actions at the pituitary level. Gen Comp Endocrinol 165:60–71. https://doi.org/10.1016/j.ygcen.2009.06.001
Zhou Y, Niu Y, Tao M et al (2010) Molecular cloning, characterization and expression of FSH and LH beta subunits from grass carp (Ctenopharyngodon idella). Fish Physiol Biochem 36:213–221. https://doi.org/10.1007/s10695-008-9223-4
Zmora N, González-Martínez D, Muñoz-Cueto J-A et al (2002) The GnRH system in the European sea bass (Dicentrarchus labrax). J Endocrinol 172:105–116. https://doi.org/10.1677/joe.0.1720105
Zmora N, Stubblefield J, Zulperi Z, Muñoz-Cueto J-A (2012) Differential and gonad stage-dependent roles of kisspeptin1 and kisspeptin2 in reproduction in the modern teleosts, Morone species. Biol Reprod 86:177. https://doi.org/10.1095/biolreprod.111.097667
Zohar Y, Muñoz-Cueto J-A, Elizur A, Kah O (2010) Neuroendocrinology of reproduction in teleost fish. Gen Comp Endocrinol 165:438–455. https://doi.org/10.1016/j.ygcen.2009.04.017
Acknowledgments
We gratefully thank to staff of Sesoko Station, Tropical Biosphere Research Center, University of the Ryukyus, Okinawa, Japan, for use of facilities. This study was supported in part by a Grant-in-Aid for Scientific Research (B) (KAKENHI, Grant number 16H05796) from the Japan Society for the Promotion of Science (JSPS) to AT and Heiwa Nakajima Foundation to AT.
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Imamura, S., Hur, SP., Takeuchi, Y. et al. The mRNA expression patterns of kisspeptins, GnRHs, and gonadotropins in the brain and pituitary gland of a tropical damselfish, Chrysiptera cyanea, during the reproductive cycle. Fish Physiol Biochem 46, 277–291 (2020). https://doi.org/10.1007/s10695-019-00715-5
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DOI: https://doi.org/10.1007/s10695-019-00715-5