The objective of the study was to elucidate mRNA expression of CYP26b1 (cytochrome P450, family 26, subfamily B, polypeptide 1) and signalling molecules ALDH1 (aldehyde dehydrogenase 1), CRABPII (cellular retinoic acid-binding protein II), RARα (retinoic acid receptor alpha) and STRA8 (stimulated by retinoic acid gene 8) in dog testis from different post-natal developmental ages. Testicular tissue samples were collected from medium-sized mixed breed dogs at different ages such as young (<4 months; N = 4), peripubertal (4–8 months; N = 3) and adult (>8 months; N = 4) were used to evaluate relative mRNA expression. Genes of RA-degrading enzyme CYP26b1, ALDH1 involved in RA synthesis and genes of carrier protein CRABPII involved in RA metabolism were turned on during the post-natal testicular development in dogs. Their expression pattern differs at different developmental ages (p < 0.05), and the levels of mRNA expression were compensated towards a normal developmental response for the sexual maturity and continuous spermatogenesis. The mRNA expression of RARα, one of the RA receptors participates in RA signalling in connection to spermatogenesis, was recorded in young and adult stages at varying degree. STRA8 is one of the responsive genes with regard to meiosis, and this functional gene product was expressed in all ages with the changing level (p < 0.01). In summary, the expression pattern of RA signalling molecules differed from young to adult ages, and it is expected that these changes are to compensate towards a normal developmental response for the sexual maturity and continuous spermatogenesis.