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Melatonin has a direct inhibitory effect on the basal and oestradiol-stimulated mitotic activity of primary spermatogonia in the testis of the frog, Rana esculenta. In this study oestradiol was used to induce spermatogonial proliferation to verify the anti-proliferative effect of melatonin. The colchicine metaphase arrest technique was used. The results obtained from in vivo experiments confirm that oestradiol increases the mitotic index of primary spermatogonia and, for the first time, indicate that melatonin has an inhibitory role on the proliferation of primary spermatogonia in the frog testis. Similar results were obtained from testes of melatonin-injected frogs that were exposed to oestradiol in vitro; in fact spermatogonia were unresponsive to hormonal stimulation. In addition, in short-term cultured testes, melatonin (at physiological concentration) interferes with the effects of oestradiol on spermatogonial proliferation, supporting the hypothesis that melatonin exerts the inhibitory effect directly via its local action on the frog gonads. Morphological observation after in vivo or in vitro melatonin treatments indicates that Leydig cells display degenerative features, whereas in adjacent germinal tubules, Sertoli cells show heterochromatic nuclei. These results indicate that melatonin may act on Leydig cells and confirm that there is a paracrine interaction between interstitial and germinal compartments. The results of the present study indicate, for the first time, that melatonin may be directly involved in the inhibitory control of spermatogonial proliferation in the testis of the frog, R. esculenta.

reproduction

Inhibition of the basal and oestradiol-stimulated mitotic activity of primary spermatogonia by melatonin in the testis of the frog, Rana esculenta, in vivo and in vitro