Gene structure, recombinant expression and function characterization of Siniperca chuatsi Fsrp-3

J Fish Biol. 2019 May;94(5):714-724. doi: 10.1111/jfb.13931. Epub 2019 Mar 1.

Abstract

A full-length complementary (c)DNA sequence encoding follistatin-related protein 3 (fsrp-3) was determined from skeletal muscle in Chinese mandarin fish Siniperca chuatsi, its molecular structure was characterised and its function suggested. The putative structure of S. chuatsi Fsrp-3 contains an N-terminal domain and two follistatin domains. Quantitative reverse-transcription (qRT)-PCR assays revealed that fsrp-3 messenger (m)RNA was differentially expressed among assayed tissues and was highly expressed in heart and intestine. fsrp-3 mRNA exhibited increasing expression from the larval to the juvenile stage (500 g). To investigate the potential function of S. chuatsi fsrp-3 in muscle growth, we constructed a Fsrp-3 prokaryotic expression system and injected the purified Fsrp-3 fusion protein into the dorsal muscle. Fsrp-3 administration significantly influenced cross-section area, satellite cell activation frequency and nuclear density of S. chuatsi muscle fibres. Following Fsrp-3 treatment, the expression of myogenic regulatory factors was up-regulated and decline in the expression of myostatin was observed. The study revealed that Fsrp-3 may affect muscle growth by regulating myogenic regulatory factor expression and antagonizing myostatin function to initiate satellite cell activation and differentiation in S. chuatsi.

Keywords: follistatin; molecular cloning; muscle growth; myogenic regulatory factor; satellite cell activation.

MeSH terms

  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • Fish Proteins / chemistry
  • Fish Proteins / genetics*
  • Fish Proteins / metabolism
  • Fishes / genetics*
  • Fishes / growth & development
  • Fishes / metabolism
  • Follistatin-Related Proteins / chemistry
  • Follistatin-Related Proteins / genetics*
  • Follistatin-Related Proteins / metabolism
  • Muscle, Skeletal / growth & development
  • Muscle, Skeletal / metabolism
  • RNA, Messenger / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • DNA, Complementary
  • Fish Proteins
  • Follistatin-Related Proteins
  • RNA, Messenger
  • Recombinant Proteins