Opposing Effects of Activin A and Follistatin on Developing Skeletal Muscle Cells

doi:10.1006/excr.1997.3575

Experimental Cell Research

Volume 233, Issue 2, 15 June 1997, Pages 350-362

https://doi.org/10.1006/excr.1997.3575 Get rights and content

Abstract

Activin and the activin-binding protein follistatin modulate a variety of biological processes and are abundant at sites of muscle development. Activin and follistatin were expressed in developing chick pectoral musclein vivoand in primary cell culture. Addition of recombinant activin inhibited muscle development in a dose-dependent manner as measured by the number of nuclei in myosin heavy chain positive cells and creatine phosphokinase activity. Conversely, follistatin potentiated muscle development. The effects of activin were found to be distinct from those of the related protein transforming growth factor (TGF) β1. Muscle development was repressed by activin at all time points investigated and did not recover with the removal of activin following a limited exposure. In contrast, while myogenic differentiation in TGFβ1 was initially repressed, muscle marker expression recovered to control levels—even in the continued presence of TGFβ1. Fibroblast growth factor (FGF) had little effect on inhibiton of muscle development caused by activin A. However, inhibition of development produced by TGFβ increased with increasing concentrations of FGF. Finally, early expression of myoD and myf5 mRNA by muscle cultures in the presence of activin and follistatin was analyzed. Activin-treated cultures expressed reduced myoD and myf5 levels at 1.5 days after plating. Myf5 levels in follistatin-treated cultures were elevated, but, surprisingly, these cultures showed a reduction in myoD levels. These data suggest that endogenously expressed activin and follistatin are important modulators of muscle development.

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More recently, depletion of myostatin b (mstn-2) promotes somatic growth and lipid metabolism in zebrafish (Gao et al., 2016). The autocrine glycoprotein follistatin (FST) antagonizes the function of MSTN as a competitive binding protein and promotes muscle growth in vivo (Nakamura et al., 1990; Link and Nishi, 1997; Amthor et al., 2004). FST overexpression in transgenic mice results in dramatic increases in muscle mass caused by hyperplasia and/or hypertrophy, with even greater effects detected in Mstn-knockout mice (Lee and McPherron, 2001).
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To promote directional breeding in important aquaculture species blunt snout bream, our research papers presents novel data on fish breeding. Our article complies with the Policy Statement for submission of manuscripts to the Genetics Section.
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Identification of a second follistatin gene in grass carp (Ctenopharyngodon idellus) and its regulatory function in myogenesis during embryogenesis
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Follistatin can antagonize the function of myostatin as a competitive binding protein and promote muscle growth in vivo. Here, we report the isolation and characterization of a second follistatin gene fst2 in grass carp (Ctenopharyngodon idellus). The grass carp fst2 cDNA was 1,376 bp in length, with an open reading frame (ORF) encoding 350 amino acid residues. A relatively low sequence identity of 78% was found between grass carp Fst2 and its paralog Fst1. Sequence and phylogenetic analyses suggest that the grass carp fst2 originated from fish-specific gene duplication. In adult fish, fst2 mRNA expression was observed in most tissues but was strongly expressed in the eyes, muscles, skin and ovary. Grass carp fst2 mRNA could be detected as early as 16 h post-fertilization (hpf), while fst1 mRNA was detected throughout embryogenesis. Using in situ hybridization, fst2 transcripts were detected in the anterior somites at 24 hpf and in the brain and posterior somites at 36 hpf. Meanwhile, fst1 mRNA was transcribed mainly in the optic vesicle and at the cephalic mesoderm at 12 hpf, in the eyes, cephalic mesoderm and at the lateral edge of most somites at 24 hpf, and mainly in the brain at 36 hpf. Furthermore, overexpression of fst2 mRNA markedly affected the formation of the embryonic midline and somite structures. Based on comparisons with fst1, our findings suggest that fst2 retained the ancestral functions of regulating muscle development and growth during embryogenesis in grass carp.
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So we suggest that granulosa cell could be the main source of FS, and FS is likely to exert actions on granulosa cells by autocrine signaling to regulate the ovary development of oysters like in other animals. Though the expression of Ca-follistatin was not intensive in adductor muscle, many studies also have shown that FS played important roles in promoting the muscle growth (Matzuk et al., 1995; Link and Nishi, 1997; Lee and McPherron, 2001). In addition, FS has been proved to perform other biological functions, such as regulating the embryonic development (Hashimoto et al., 1997) and inhibiting the replication of tumor cell (Miyanaga et al., 1993).
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Treatment with activin A inhibited multinucleated myotube formation in primary chicken muscle cell cultures in a dose-dependent manner, whereas follistatin enhanced myotube formation (Link and Nishi, 1997). Since inhibin/activin βA and its receptors are expressed in skeletal muscle (Hildén et al., 1994; Link and Nishi, 1997; Tuuri et al., 1994), activin A appears to be an autocrine negative regulator of myogenesis. This is supported by in vivo implant experiment in which beads coated with activin A were implanted into the chick limb bud and transiently downregulated the expression of Pax-3 and MyoD, transcription factors that are responsible for myoblast growth and differentiation (He et al., 2005).
Activins, members of the TGF-β family, are multifunctional growth and differentiation factors. Activins regulate glucose/energy metabolism by promoting the differentiation of insulin-producing and -responsive cells, and regulating function of the differentiated cells. In the pancreas, activins stimulate the differentiation of β cells and secretion of insulin, which enables the cells to respond to glucose uptake efficiently. By contrast, in the liver, skeletal muscle and white adipose tissue, activins exert negative regulation on organogenesis, which leads to impaired insulin sensitivity. Activins induce the phenotypic switch of macrophages from the M1 to M2 phenotypes, which reduces inflammation. Since adipose inflammation is closely associated with insulin resistance and the onset of type 2 diabetes, activins may improve insulin resistance through their anti-inflammatory activity. Because activins modulate events involved in insulin sensitivity in a tissue-dependent manner, the activities of activins should be locally regulated to improve whole-body insulin responsiveness. Thus, activins or activin inhibitors may be effective as therapeutic agents for metabolic syndrome.
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^☆: M. A. Bronner-Fraser, Ed.

¹: To whom correspondence should be addressed. Fax: (503) 494-4253. E-mail: [email protected].

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Regular ArticleOpposing Effects of Activin A and Follistatin on Developing Skeletal Muscle Cells☆

Abstract

Methods Enzymol.

Neuron

Dev. Biol.

Anal. Biochem.

Curr. Opin. Genet. Dev.

Dev. Biol.

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J. Biol. Chem.

Mol. Cell. Endocrinol.

FEBS Lett.

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Exp. Cell Res.

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Proc. Natl. Acad. Sci. USA

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Trends Genet.

Development

J. Biophys. Biochem. Cytol.

J. Cell Biol.

Development

Development

Regular Article
Opposing Effects of Activin A and Follistatin on Developing Skeletal Muscle Cells☆