Sexual dimorphic expression of dnd in germ cells during sex reversal and its requirement for primordial germ cell survival in protogynous hermaphroditic grouper

Comp Biochem Physiol B Biochem Mol Biol. 2017 Jun:208-209:47-57. doi: 10.1016/j.cbpb.2017.04.003. Epub 2017 Apr 8.

Abstract

Dead end (dnd), vertebrate-specific germ cell marker, had been demonstrated to be essential for primordial germ cell (PGC) migration and survival, and the link between PGC number and sex change had been revealed in some teleost species, but little is known about dnd in hermaphroditic vertebrates. In the present study, a protogynous hermaphroditic orange-spotted grouper (Epinephelus coioides) dnd homologue (Ecdnd) was identified and characterized. Quantitative real-time PCR and in situ hybridization analysis revealed a dynamic and sexually dimorphic expression pattern in PGCs and germ cells of gonads. During sex changing, the Ecdnd transcript sharply increased in early transitional gonad, reached the highest level at late transitional gonad stage, and decreased after testis maturation. Visualization of zebrafish PGCs by injecting with RFP-Ecdnd-3'UTR RNA and GFP-zfnanos3-3'UTR RNA confirmed importance of Ecdnd 3'UTR for the PGC distribution. In addition, knockdown of EcDnd by using antisense morpholinos (MO) caused the ablation of PGCs in orange-spotted grouper. Therefore, the current data indicate that Ecdnd is essential for PGCs survival and may serve as a useful germ cell marker during gametogenesis in hermaphroditic grouper.

Keywords: Epinephelus coioides; Morpholino; PGC; Sex change; dnd.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Female
  • Fish Proteins / genetics
  • Fish Proteins / metabolism*
  • Gene Expression Regulation
  • Gene Expression Regulation, Developmental
  • Germ Cells / cytology*
  • Germ Cells / metabolism
  • Gonads / embryology
  • Gonads / metabolism*
  • Hermaphroditic Organisms / genetics
  • Hermaphroditic Organisms / growth & development
  • Hermaphroditic Organisms / metabolism*
  • In Situ Hybridization
  • Male
  • Phylogeny
  • Sequence Homology, Amino Acid
  • Sex Characteristics*
  • Sex Differentiation*
  • Zebrafish / genetics
  • Zebrafish / metabolism*

Substances

  • Fish Proteins