Isolation and Expression Analysis of the Canine Insulin-Like Factor 3 Gene

Anne Truong, Natalia Bogatcheva, Claude Schelling, Gaudenz Dolf, Alexander I. Agoulnik

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The insulin-like factor 3 (INSL3 or relaxin-like factor) is a hormone produced mainly in gonadal tissues in males and females. Deletion of INSL3 or its receptor in male mice leads to the undescended testes, or cryptorchidism. Here we describe an isolation and analysis of full-length canine INSL3 gene. The INSL3 gene is composed of two exons within a small genomic region. Putative translation of the isolated cDNA yields 132 amino acid preproINSL3 that has the domain structure characteristic for the insulin-relaxin peptide superfamily with a well-conserved receptor-binding domain. Northern blot hybridization showed stronger expression of INSL3 in testis than in ovary. Reverse transcription-polymerase chain reaction analysis of the INSL3 expression revealed a minor splice variant of INSL3 potentially encoding 105 amino acids peptide. We established that the medium, conditioned with recombinant canine INSL3, produced from the full-length cDNA, but not from the minor splice variant, activated human GREAT/LGR8 receptor in vitro. In addition to the functional allele of INSL3, genomic DNA of one of the analyzed dogs contained an intronless nonexpressed pseudogene of INSL3. We isolated canine INSL3 promoter and showed that its activity was strongly mediated by steroidogenic factor-1 in vitro. Using site-specific mutagenesis, we identified a well-conserved steroidogenic factor-1 binding site within canine INSL3 promoter.

Original languageEnglish (US)
Pages (from-to)1658-1664
Number of pages7
JournalBiology of Reproduction
Volume69
Issue number5
DOIs
StatePublished - Nov 2003
Externally publishedYes

Fingerprint

Canidae
Steroidogenic Factor 1
Insulin
Cryptorchidism
Genes
Complementary DNA
Relaxin
Amino Acids
Peptides
Pseudogenes
Conditioned Culture Medium
Site-Directed Mutagenesis
Northern Blotting
Reverse Transcription
Testis
Ovary
Exons
Alleles
Binding Sites
Hormones

Keywords

  • Developmental biology
  • Gene regulation
  • Male reproductive tract
  • Relaxin
  • Testis

ASJC Scopus subject areas

  • Cell Biology
  • Developmental Biology
  • Embryology

Cite this

Isolation and Expression Analysis of the Canine Insulin-Like Factor 3 Gene. / Truong, Anne; Bogatcheva, Natalia; Schelling, Claude; Dolf, Gaudenz; Agoulnik, Alexander I.

In: Biology of Reproduction, Vol. 69, No. 5, 11.2003, p. 1658-1664.

Research output: Contribution to journalArticle

Truong, Anne ; Bogatcheva, Natalia ; Schelling, Claude ; Dolf, Gaudenz ; Agoulnik, Alexander I. / Isolation and Expression Analysis of the Canine Insulin-Like Factor 3 Gene. In: Biology of Reproduction. 2003 ; Vol. 69, No. 5. pp. 1658-1664.
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