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<p>Sarcospan (SSPN) is an integral component of the dystrophin-glycoprotein complex (DGC) that contributes significantly to mammalian muscle development. However, its function remains largely unknown in teleosts, especially in flatfish. In this study, we firstly cloned and characterized the <i>sspn</i> gene from <i>Cynoglossus semilaevis</i> (<i>Cs_sspn</i>). The full-length cDNA was 1 735 bp, with a 669-bp open reading frame (ORF), encoding 222 amino acids. The predicted SSPN protein (<i>Cs</i>_SSPN) comprised four transmembrane domains and showed a close relationship to the tetraspanin family. To determine the subcellular localization of <i>Cs</i>_SSPN, HEK293T cells were used as an in vitro model, revealing predominant plasma membrane localization with minor cytoplasmic distribution. Phylogenetic analysis showed that <i>sspn</i> genes from Pleuronectiformes clustered into a distinct clade, and branch-site model analysis detected four positively selected sites located in the extracellular (64T), transmembrane (111V), and intracellular (30G and 200D) domains, supporting a proposed model of adaptive evolution in flatfishes that requires further validation. Additionally, <i>sspn</i> mRNA transcripts were significantly expressed in muscle tissue (eyed side and blind side), suggesting an important role in muscle development. Furthermore, silencing <i>Cs_sspn</i> through RNA interference (RNAi) dramatically reduced the expression levels of six muscle development-related genes including α-sarcoglycan (<i>sgca</i>), β-sarcoglycan (<i>sgcb</i>), δ-sarcoglycan (<i>sgcd</i>), γ-sarcoglycan (<i>sgcg</i>), dystroglycan (<i>dag1</i>), and dystrophin (<i>dmd</i>). All these results confirm the indispensable role of <i>sspn</i> in the regulation of muscle development in <i>C. semilaevis</i>.</p>

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Characterization, expression, and functional analysis of sarcospan (SSPN) in Chinese tongue sole: Cynoglossus semilaevis

  • 期刊论文

  • 发表日期：

    2026年5月30日

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  • Jing Yu,
  • Yijing Yang,
  • Tianwei Liu,
  • Kun Huang,
  • An Xu,
  • Li Gong,
  • Jing Liu,
  • Fenghui Li,
  • Denghui Zhu,
  • Liqin Liu,
  • Zhenming Lü

摘要

Sarcospan (SSPN) is an integral component of the dystrophin-glycoprotein complex (DGC) that contributes significantly to mammalian muscle development. However, its function remains largely unknown in teleosts, especially in flatfish. In this study, we firstly cloned and characterized the sspn gene from Cynoglossus semilaevis (Cs_sspn). The full-length cDNA was 1 735 bp, with a 669-bp open reading frame (ORF), encoding 222 amino acids. The predicted SSPN protein (Cs_SSPN) comprised four transmembrane domains and showed a close relationship to the tetraspanin family. To determine the subcellular localization of Cs_SSPN, HEK293T cells were used as an in vitro model, revealing predominant plasma membrane localization with minor cytoplasmic distribution. Phylogenetic analysis showed that sspn genes from Pleuronectiformes clustered into a distinct clade, and branch-site model analysis detected four positively selected sites located in the extracellular (64T), transmembrane (111V), and intracellular (30G and 200D) domains, supporting a proposed model of adaptive evolution in flatfishes that requires further validation. Additionally, sspn mRNA transcripts were significantly expressed in muscle tissue (eyed side and blind side), suggesting an important role in muscle development. Furthermore, silencing Cs_sspn through RNA interference (RNAi) dramatically reduced the expression levels of six muscle development-related genes including α-sarcoglycan (sgca), β-sarcoglycan (sgcb), δ-sarcoglycan (sgcd), γ-sarcoglycan (sgcg), dystroglycan (dag1), and dystrophin (dmd). All these results confirm the indispensable role of sspn in the regulation of muscle development in C. semilaevis.