Gonadal sex differentiation in frogs: how testes become shorter than ovaries.

Testis differentiation in anuran amphibians is the result of two opposing processes: degeneration of the distal part, and development of the proximal part, which becomes a functional male gonad. Undifferentiated gonad differentiates directly into a testis without a transition phase. We described the morphology of developing testes in Rana temporaria and Hyla arborea, and made careful histology and ultrastructure in Pelophylax lessonae. The developing testis was divided into 10 stages (I-III, undifferentiated gonad, IV-X, testis). The earliest morphological symptoms of testis differentiation were observed in 4- to 5-week-old tadpoles at Gosner stage 27-28. At that time an undifferentiated gonad, composed of 6-9 metameres, differentiates into a testis. The proximal metameres (2-3 in the right gonad and 3-4 in the left one) differentiate into a functional testis, while the distal ones degenerate. The difference between left and right gonad size is maintained until adulthood (stage X). Degeneration of the distal part progresses along the posterior-anterior gradient and starts at stage IV. It affects first the germ cells with accompanying precursor Sertoli cells, and then the mesenchymal cells and fibroblasts. Finally the external epithelium forms a "sleeve" around the almost empty distal part. The total length of the testes stays the same until stage VIII at Gosner stage 41 (age 74-148 days). Active spermatogenesis starts at stage IX (juveniles after their first hibernation), during which the distal part eventually disappears and the proximal part starts growing considerably due to progressing spermatogenesis.