Effects of sex steroids on expression of myostatin in rare minnow, Gobiocypris rarus
Highlights
► MSTN1 named GrMSTN was isolated from rare minnow, Gobiocypris rarus. ► GrMSTN is maternal factor and expressed ubiquitously in adult tissues in rare minnow. ► 1 μg/L sex steroids significantly decrease body growth and increase MSTN expression. ► It is suggested that sex steroids can affect body growth through MSTN expression.
Introduction
Myostatin (MSTN), a member of transforming growth factor (TGF) β superfamily, negatively regulates muscular development in mammals (Grobet et al., 1997, Kambadur et al., 1997, Lee and McPherron, 2001, McPherron and Lee, 1997, McPherron et al., 1997). Fish have two MSTN genes, MSTN1 and MSTN2, involved in muscular development (Acosta et al., 2005, Amali et al., 2004, Amali et al., 2008, Biga et al., 2005, Kerr et al., 2005). Knockdown of MSTN1 induced giant phenotype and up-regulation of muscular specific genes in zebrafish (Danio rerio) (Acosta et al., 2005, Amali et al., 2004). Transgenic medaka (Oryzias latipes) with expression of dominant-negative MSTN1 exclusively in muscle increased skeleton muscle fibers at adult (Sawatari et al., 2010). Overexpression of MSTN2 in zebrafish led to muscle loss and expression of dystrophin associated protein complex, which resulted in muscle dystrophy (Amali et al., 2008).
Hormones and environmental factors can regulate the expression of MSTN in fish but the effects are different with species. Overexpression of growth hormone (GH) or administration of recombinant bovine growth hormone (rbGH) decreased MSTN2 level in coho salmon (Oncorhynchus kisutch) (Roberts et al., 2004) and rainbow trout (Oncorhynchus mykiss) (Biga et al., 2004). Overexpression of GH increased MSTN1 level in rainbow trout (Biga et al., 2004), but rbGH administration did not change MSTN1 level in coho salmon (Roberts et al., 2004). GH decreased MSTN level in giant danio (Danio aequipinnatus) while it increased MSTN level in zebrafish (Biga and Meyer, 2009). Administration of dexamethasone, a glucocorticoid resulted in loss of body weight and muscle atrophy associated with induction of MSTN expression in channel catfish (Ictalurus punctatus) (Weber et al., 2005). However, cortisol reduced MSTN level in tilapia (Oreochromis mossambicus) larvae (Rodgers et al., 2003). Fasting and refeeding did not affect MSTN mRNA levels in adult tilapia (Rodgers et al., 2003) and rainbow trout (Chauvigne et al., 2003). By contrast, larval MSTN mRNA levels were sometimes elevated after a short-term fast and were consistently reduced with prolonged fasting (Rodgers et al., 2003). In chronically overcrowded environment, both depression of body growth and a diminished level of MSTN mRNA in the adult zebrafish were observed (Vianello et al., 2003).
Sex steroids are important for sex differentiation and also for muscular development. Androgen can increase muscle in man and woman, and androgenic compounds have been misused in athletes to increase their muscle mass, skeletal muscle strength and performance (Miller, 2009). However, the effects of androgen on MSTN expression arouse controversy. Some people found that testosterone did not change MSTN expression in human (Homo sapiens) (Kvorning et al., 2007, Lewis et al., 2007), the others reported that testosterone increased MSTN expression (Lakshman et al., 2009). Estrogen replacement attenuated muscle growth of young ovariectomized rat (Rattus rattus) (Piccone et al., 2005). Transient (1 week) usage of estrogen decreased insulin-like growth factor-1 (IGF1) and increased MSTN expression, however long time (5 week) administration of estrogen did not apparently influence the expression of IGF1 and MSTN in rat (Tsai et al., 2007).
Some environmental endocrine disruptors can mimic the effects of sex steroid and affect growth of fish. Androgen can increase muscle mass in weakfish (Cynoscion regalis) (Connaughton and Taylor, 1995) and promote growth of tilapia (Sparks et al., 2003) and coho salmon (Larsen et al., 2004). 17alpha-ethinylestradiol (EE2) impairs growth, expression of IGF1 and IGF2 in brook trout (Salvelinus fontinalis) (Schafhauser-Smith and Benfey, 2003) and tilapia (Shved et al., 2009). However, whether sex steroids affect fish growth through MSTN has not been reported.
Rare minnow (Gobiocypris rarus), a small native cyprinid fish, is currently used in aquatic toxicology in China because of its sensitivity to environmental endocrine disruptors (Ma et al., 2007, Zha et al., 2008, Zhong et al., 2005). Here, we report the isolation of MSTN from rare minnow and the effects of androgen/estrogen on body growth and MSTN mRNA levels in rare minnow.
Section snippets
Fish
The fish of rare minnow were obtained from Institute of Hydrobiology, Chinese Academy of Sciences, Wuhan, China, and were kept in our laboratory. The fish were fed twice a day with commercial diet in the environment of ambient temperature at 28 °C and light cycles of 14 hour light and 10 hour dark. The embryos were obtained by artificial fertilization with eggs and milt from normal adult fish and cultured in water at 28 °C.
The work described in this article was carried out in accordance with The
Sequence analysis of rare minnow MSTN
The sequence of rare minnow MSTN has been submitted to GenBank (ID: FJ482232) and is named as GrMSTN. The full length of GrMSTN cDNA is 2184 bp containing an open reading frame (ORF) of 1128 bp encoding 375 amino acids, the 5′ untranslated region (UTR) of 88 bp and the 3′ UTR including poly(A) tail of 968 bp. The deduced GrMSTN protein has a signal peptide (residues 1–22) predicted by the SignalP V4.0 and two conserved domains, TGF-β propeptide domain (residues 23–262) and TGF-β domain (mature MSTN
Discussion
We isolated MSTN cDNA named as GrMSTN from rare minnow. Phylogenetic analysis indicates that GrMSTN is the homologue of zebrafish MSTN1. Two MSTN genes exist in fish genome. Duplicated MSTN in fish is the result of whole genome duplication occurring in the ancestor of modern bony fish shortly after separation with tetrapod, while salmonids have two MSTN1 and two MSTN2 because of an additional event of whole genome duplication occurring in the salmonids (Kerr et al., 2005).
GrMSTN is expressed
Acknowledgments
This work was supported by the Open grant of State Key Laboratory of Freshwater Ecology and Biotechnology, Chinese Academy of Sciences (2005FB18).
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Research progresses of myostatin in fish muscle
2022, Zhongshan Daxue Xuebao/Acta Scientiarum Natralium Universitatis Sunyatseni
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These authors contribute equally.