Cite this paper:
WEN Haishen, SI Yufeng, ZHANG Yuanqing, HE Feng, LI Jifang. Cloning and expression of follistatin gene in half-smooth tongue sole Cynoglossus semilaevis during the reproduction cycle[J]. Journal of Oceanology and Limnology, 2015, 33(2): 299-308

Cloning and expression of follistatin gene in half-smooth tongue sole Cynoglossus semilaevis during the reproduction cycle

WEN Haishen1,2, SI Yufeng1, ZHANG Yuanqing1, HE Feng1, LI Jifang1
1 College of Fisheries, Ocean University of China, Qingdao 266003, China;
2 College of Animal Science and Technology, Inner Mongolia Nationality University, Tongliao 028000, China
Abstract:
Follistatin (FST) is a monomeric glycoprotein highly enriched in cysteines and belongs to TGF-β superfamily. FST can suppress the secretion of follicle-stimulating hormone and plays a vital role in the reproduction of vertebrates. We used rapid amplification of cDNA ends technology to clone the FST gene of half-smooth tongue sole, Cynoglossus semilaevis. We characterized its phylogenetic context and expression patterns to elucidate its function in the breeding season. The full-length sequence of FST is 1 455 bp and encodes a protein of 321 amino acids. We investigated the expression pattern of FST in C. semilaevis at different stages of reproduction using reverse transcription-polymerase chain reaction (RTPCR). FST mRNA was expressed in all 13 tissues analyzed, and was expressed at high levels in gonad and at slightly lower levels in gill and brain. During the reproductive cycle of C. semilaevis, the transcript level of FST was the highest in the perinucleolus stage, decreased in the primary yolk stage, slightly increased in the tertiary yolk stage, and then reduced to a minimal level in the atretic follicles stage of the ovary. We concluded that FST suppressed follicle-stimulating hormone, which stimulated oocyte development. However, no significant variation was observed across all stages of testis development, although the expression level in the spermatogenesis stage was relatively low, which may result from the regulation of FST by aromatase.
Key words:    Cynoglossus semilaevis|follistatin (FST)|cloning|gene expression   
Received: 2014-03-14   Revised: 2014-05-06
Tools
PDF (456 KB) Free
Print this page
Add to favorites
Email this article to others
Authors
Articles by WEN Haishen
Articles by SI Yufeng
Articles by ZHANG Yuanqing
Articles by HE Feng
Articles by LI Jifang
References:
Altschul S F, Gish W, Miller W, Myers E W, Lipman D J. 1990. Basic local alignment search tool. J. Mol. Biol., 215 (3): 403-410.
Aroua S, Maugars G, Jeng S R, Chang C F, Weltzien F A, Rousseau K, Dufour S. 2012. Pituitary gonadotropins FSH and LH are oppositely regulated by the activin/ follistatin system in a basal teleost, the eel. General and Comparative Endocrinology, 175 (1): 82-91.
Bauer H, Meier A, Hild M, Stachel S, Economides A, Hazelett D, Harland R M, Hammerschmidt M. 1998. Follistatin and noggin are excluded from the zebrafish organizer. Dev. Biol., 204 (2): 488-507.
Cash J N, Angerman E B, Kattamuri C, Nolan K, Zhao H, Sidis Y, Keutmann H T, Thompson T B. 2012. Structure of myostatin follistatin-like 3 N-terminal domains of follistatin-type molecules exhibit alternate modes of binding. Journal of Biological Chemistry, 287 (2): 1 043- 1 053.
Chen C F, Wen H S, Chen X Y, Shi B, Shi D, Yang Y P. 2010a. Studies on ovarian development and spawn type of cultured half-smooth tongue sole, Cynoglossus semilaevis. Marine Sciences, 8 (34): 29-34. (in Chinese with English abstract)
Chen C F, Wen H S, He F, Dong S L. 2009. Programming design of degenerate primers and cloning half-smooth tongue sole Cynoglossus semilaevis CYP17 gene. Periodical of Ocean University of China, 6 (39): 1 213- 1 218. (in Chinese with English abstract)
Chen C F, Wen H S, Wang Z P, He F, Zhang J R, Chen X Y, Jin G X, Shi B, Shi D, Yang Y P, Li J F, Qi B X, Li N. 2010b. Cloning and expression of P450c17-I
(17α-hydroxylase/17,20-lyase) in brain and ovary during gonad development in Cynoglossus semilaevis. Fish Physiology and Biochemistry, 4 (36): 1 001-1 012.
Chen C F, Wen H S. 2012. The characteristic analysis of P450c17s mRNA expression during the reproductive cycle in male Cynoglossus semilaevis Günther. Journal of Fisheries of China, 36 (7): 1 019-1 025. (in Chinese with English abstract)
Chen S L, Zhang G J, Shao C W, Huang Q F, Liu G, Zhang P, Song W T, An N, Domitille C, Jean-Nicolas V, Hong Y H, Li Q Y, Sha Z X, Zhou H L, Xie M S, Yu Q L, Liu Y, Xiang H, Wang N, Wu K, Yang C G, Zhou Q, Liao X L, Yang L F, Hu Q M, Zhang J L, Meng L, Jin L J, Tian Y S, Lian J M, Yang J F, Miao G D, Liu S S, Liang Z, Yan F, Li Y Z, Sun B, Zhang H, Zhang J, Zhu Y, Du M, Zhao Y W, Manfred S, Tang Q S, Wang J. 2014. Whole-genome sequence of a flatfish provides insights into ZW sex chromosome evolution and adaptation to a benthic lifestyle. Nature genetics, 46 : 253-260.
Chen X Y, Wen H S, He F, Li J F, Chen C F, Yang Y P, Zhang J R, Jin G X, Shi B. 2010c. Cloning of FSHR and expression analysis during the reproductive cycle in female Cynoglossus semilaevis Günther. Journal of Fisheries of China, 9 (34): 1 309-1 318. (in Chinese with English abstract)
Chen, S L, Li J, Deng S P, Tian Y S, Wang Q Y, Zhuang Z M, Sha Z X, Xu J Y. 2007. Isolation of female-specific AFLP markers and molecular identification of genetic sex in half-smooth tongue sole (Cynoglossus semilaevis). Marine Biotechnology, 9 (2): 273-280.
de Santis C, Gomes G B, Jerry D R. 2012. Abundance of myostatin gene transcripts and their correlation with muscle hypertrophy during the development of barramundi, Lates calcarifer. Comparative Biochemistry and Physiology Part B : Biochemistry and Molecular Biology, 163 (1): 101-107.
Dong X L, Chen S L, Ji X S, Shao C W. 2011. Molecular cloning, characterization and expression analysis of Sox9a and Foxl2 genes in half-smooth tongue sole (Cynoglossus semilaevis). Acta Oceanologica Sinica, 30 (1): 68-77.
Ghaia S, Mongaa R, Mohantyb T K, Chauhanc M S, Singh D. 2010. Tissue-specific promoter methylation coincides with Cyp19 gene expression in buffalo (Bubalus bubalis) placenta of different stages of gestation. Gen. Comp. Endocr., 2 (169): 182-189.
Gregory D J, Waldbieser G C, Bosworth B G. 2004. Cloning and characterization of myogenic regulatory genes in three Ictalurid species. Anim. Genet., 6 (35): 425-430.
Ji X S, Chen S L, Jiang Y L, Xu T J, Yang J F, Tian Y S. 2011. Growth differences and differential expression analysis of pituitary adenylate cyclase activating polypeptide (PACA P) and growth hormone-releasing hormone (GHRH) between the sexes in half-smooth tongue sole Cynoglossus semilaevis. General and Comparative Endocrinology, 170 (1): 99-109.
Kocams H, Gulmez N, Aslan S, Nazli M. 2004. Follistatin alters myostatin gene expression in C 2 C 12 muscle cells. Acta. Vet. Hung., 2 (52): 135-141.
Li S J, Bai J J, Ye X, Lao H H, Jian Q. 2007. Cloning and analysis of largemouth bass (Micropterus salmoides) follistatin cDNA and its expression in Escherichia coli. Journal of Agricultural Biotechnology, 15 (5): 783-788. (in Chinese with English abstract)
Liu Q H, Tan X G, Xu Y L, Zhang P J, Xu P. 2007. Cloning and sequence analysis of Paralichthys olivaceus Follistatin cDNA. Journal of Zhejiang University (Agriculture and Life Sciences), 6 (33): 621-625. (in Chinese with English abstract)
Liu S S, Sun B, Liang Z, Zhang J, Chen S L. 2013. Cloning and expression of anti-Müllerian hormone gene in halfsmooth tongue-sole, Cynoglossus semilaevis. Journal of Fishery Sciences of China, 20 (1): 35-43. (in Chinese with English abstract)
Liu Z F, Xue L Y, Shen W L, Ying J, Zhang Z H. 2014. Spatiotemporal expression pattern and fasting response of follistatin gene in teleost Larimichthys crocea. Genes & Genomics, 36 (2): 205-214.
Ma Q, Liu S F, Zhuang Z M, Lin L, Sun Z Z, Liu C L, Ma H, Su Y Q, Tang Q S. 2012a. Genomic structure, polymorphism and expression analysis of the growth hormone (GH) gene in female and male Half-smooth tongue sole (Cynoglossus semilaevis). Gene, 493 (1): 92- 104.
Ma Q, Liu S F, Zhuang Z M, Sun Z Z, Liu C L, Tang Q S. 2012b. The co-existence of two growth hormone receptors and their differential expression profi les between female and male tongue sole (Cynoglossus semilaevis). Gene, 511 (2): 341-352.
Nakamura T, Takio K, Eto Y, Shibai H, Titani K, Sugino H. 1990. Activin-binding protein from rat ovary is follistatin. Science, 247 : 836-838.
Ni J B, Zeng Z, Han G D, Huang H Q, Ke C H. 2012. Cloning and characterization of the follistatin gene from (Crassostrea angulate) and its expression during the reproductive cycle. Comparative Biochemistry and Physiology Part B : Biochemistry and Molecular Biology, 163 (2): 246-253.
Nicol B, Yano A, Jouanno E, Guérin A, Fostier A, Guiguen Y. 2013. Follistatin is an early player in rainbow trout ovarian differentiation and is both colocalized with aromatase and regulated by the Wnt pathway. Sexual Development, 7 (5): 267-276.
Patel K. 1998. Follistatin. International Journal of Biochemistry & Cell Biology, 10 (30): 1 087-1 093.
Phillips D J, de Kretser D M. 1998. Follistatin: a multifunctional regulatory protein. F ront. N euroendocrin., 4 (19): 287-322.
Robertsona D M, Klein R, de Vos F L, McLachlan R I, Wettenhall R E H, Hearn M T W, Burger H G, de Kretser D M. 1987. The isolation of polypeptides with FSH suppressing activity from bovine follicular fluid which are structurally different to inhibin. Biochem. Bioph. Res. Co., 2 (149): 744-749.
Sadatsuki M, Tsutsumi O, Yamada R, Muramatsu M, Taketani Y. 1993. Local regulatory effects of Activin A and follistatin on meiotic maturation of rat oocytes. Biochem. Bioph. Res. Co., 1 (196): 388-395.
Saitou N, Nei M. 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol. Biol. Evol., 4 (4): 406-425.
Sidis Y, Mukherjee A, Keutmann H, Delbaere A, Sadatsuki M, Schneyer A. 2006. Biological activity of follistatin isoforms and Follistatin-Like-3 is dependent on differential cell surface binding and specifi city for activin, myostatin, and bone morphogenetic proteins. Endocrinology, 7 (147): 3 586-3 597.
Sidis Y, Schneyer A L, Sluss P M, Johnson L N, Keutmann H T. 2001. Follistatin: essential role for the N-terminal domain in activin binding and neutralization. The Journal of Biological Chemistry, 276 : 17 718-17 726.
Simpson E R, Clyne C, Rubin G, Boon W C, Robertson K, Britt K, Speed C, Jones M. 2002. Aromatase—a brief overview. Annu. Rev. Physiol., 64 : 93-127.
Song W T, Li Y Z, Zhao Y W, Liu Y, Niu Y Z, Pang R Y, Miao G D, Liao X L, Shao C W, Gao F T, Chen S L. 2012. Construction of a high-density microsatellite genetic linkage map and mapping of sexual and growth-related traits in half-smooth tongue sole (Cynoglossus semilaevis). PloS O ne, 7 (12): e52097.
Sugino K, Kurosawa N, Nakamura T, Takio K, Shimasaki S, Ling N, Titani K, Sugino H. 1993. Molecular heterogeneity of follistatin, an activin-binding protein. Higher affi nity of the carboxyl-terminal truncated forms for heparan sulfate proteoglycans on the ovarian granulosa cell. Journal of Biological Chemistry, 268 : 15 579-15 587.
Tsuchita K. 2005. Regulation of skeletal muscle mass and adipose tissue mass by follistatin and follistatin-related gene (FLRG) and development of novel polypeptides as medical drugs. Seikagaku, 5 (77): 440-443.
Ueno N, Ling N, Ying S Y, Esch F, Shimasaki S, Guillemin R. 1987. Isolation and partial characterization of follistatin: a single-chain Mr 35,000 monomeric protein that inhibits the release of follicle-stimulating hormone. Proc. Natl. Acad. Sci., U. S. A, 84 : 8 282-8 286.
Yuen C, Ge W. 2004. Follistatin suppresses FSHβ but increases LHβ expression in the goldfish — evidence for an activinmediated autocrine/paracrine system in fish pituitary. Gen. Comp. Endocr., 1 35 (1): 108-115.
Zhang J R. 2009. Studies on Reproduction Physiological Function of the Estrogen Receptor and Androgen Receptor in Cultured Male Cynoglossus semilaevis Günther. Doctoral dissertation, Ocean University of China. (in Chinese with English abstract)
Zhong S S, Jiang X Y, Sun C F, Zou S M. 2013. Identification of a second follistatin gene in grass carp (Ctenopharyngodon idellus) and its regulatory function in myogenesis during embryogenesis. Gen. Comp. Endocr., 185 (1): 19-27.
Copyright © Haiyang Xuebao