The Harderian gland (Hg), the only gland found in the orbit of the frog Rana esculenta L., 1758, probably plays a role in orbital lubrication. The secretory activity of the Hg is seasonal, showing the highest activity in summer. There is little information on Hg gene expression; previously, we identified a mRNA named harderin, whose deduced protein has no homology with other proteins. Differential expression of the harderin transcript between the sexes expressed during the annual cycle implies sexual dimorphism. RT–PCR (reverse transcription – polymerase chain reaction) analysis, revealed that harderin is expressed during the entire year in the Hg of both sexes. It shows a higher level of expression in the female glands than that of male glands. Two peaks of expression, in February and in June, were observed in the female glands, while only the February peak was observed in those of males. These observations were supported by in situ hybridization. Experiments involving gonadectomy and (or) hormonal replacement therapy showed a significant decrease in harderin in the Hg of females; this effect is prevented by estradiol (testosterone had no effect), while ICI (antiestrogen) counteracts the hormonal prevention, suggesting that this sexual dimorphism is under estradiol control. The constant expression of harderin mRNA during the year suggests a probable constitutive role for this molecule.