Molecular cloning and localization of caprine relaxin-like factor (RLF) mRNA within the goat testis

The relaxin-like factor (RLF) is one of the insulin-like molecules, which also includes insulin, insulin-like growth factor I and II, placentin, and relaxin. Employing RT- and RACE-PCR on RNA isolated from goat testicular tissue, we report the cloning and nucleic acid sequence of goat RLF. The caprine RLF cDNA coding sequence consisted of 396 base pairs encoding a peptide of 131 amino acids. Caprine RLF showed the highest homology in nucleic acid and amino acid sequence with bovine and sheep RLF, suggesting conservation of the RLF gene among ruminants. Northern blot analysis revealed a single 0.9 kb RLF transcript expressed in the goat testis but not in the epididymis, liver, or muscle tissue. Only a single goat RLF gene is present in the goat genome as determined by Southern blot analysis. Employing nonradioactive in situ hybridization for goat RLF mRNA and immunohistochemistry for 3beta-hydroxysteroid dehydrogenase (3beta-HSD) and 17alpha-hydroxylase, we identified the Leydig cells as the sole source of RLF mRNA in the goat testis.