Cloning, tissue distribution and methylation analyses of Lhx9 in Chinese tongue sole (Cynoglossus semilaevis)

Gene. 2019 Apr 5:691:176-184. doi: 10.1016/j.gene.2018.12.025. Epub 2018 Dec 21.

Abstract

Lhx9 is a LIM-homeodomain protein related to gonad development and sex reversal. In this study, we cloned and characterized CS-Lhx9 in the gonads of the Chinese tongue sole (Cynoglossus semilaevis). The full-length cDNA of CS-Lhx9 was 3123 bp, including an ORF of 1149 bp encoding 383 amino acids which contains two LIM domains and a homeobox domain. CS-Lhx9 transcripts were primarily observed in the testis of male and neomale at 1 yah, but nearly undetectable in the ovary. During the development of gonad, CS-Lhx9 exhibited an increasing trend and appeared to reach its peak value of expression in testis at 1 yah. In situ hybridization was performed in male and neomale gonads at 210 dah and 1 yah. The results showed strong CS-Lhx9 signals in the spermatids and spermatozoa (germ cells). Methylation level of CS-Lhx9 promoter was higher in female and lower in male and neomale gonads, showing a negative correlation with CS-Lhx9 expression.

Keywords: Expression; Gonad; ISH; Lhx9; Methylation.

MeSH terms

  • Animals
  • Cloning, Molecular / methods*
  • DNA Methylation*
  • Epigenesis, Genetic
  • Female
  • Fish Proteins / chemistry
  • Fish Proteins / genetics
  • Fish Proteins / metabolism
  • Flatfishes / genetics*
  • Flatfishes / growth & development
  • Gene Expression Regulation, Developmental
  • Male
  • Open Reading Frames
  • Organ Specificity
  • Ovary / metabolism
  • Promoter Regions, Genetic
  • Protein Domains
  • Sex Characteristics
  • Testis / metabolism
  • Tissue Distribution
  • Transcription Factors / chemistry
  • Transcription Factors / genetics*
  • Transcription Factors / metabolism*

Substances

  • Fish Proteins
  • Transcription Factors