Androgen Biosynthesis and Secretion in Developing Xenopus laevis

In the frog, Xenopus laevis, castration or anti-androgen treatment during the late tadpole or early juvenile stages blocks masculinization of vocal neuroeffectors in males while exogenous androgen or a testicular transplant masculinizes vocal neuroeffectors in females. To elucidate the relation between androgen secretion and the process of masculinization, we measured androgen by radioimmunoassay in adults, juveniles, and tadpoles. In adult females, values for both testosterone (T) and dihydrotestosterone (DHT) were essentially identical during the winter and the summer while in males, summer values for both androgens were 7 to 8x winter values. For DHT, circulating values in males significantly exceeded those in females in both winter and summer while for T, values differed significantly only in summer. Sex differences in circulating androgen arise at late juvenile stages (PM4 and PM5); T values are higher in males than in females and higher, for both sexes, in winter than in summer. During early juvenile stages (PM1-PM3), there were no seasonal or sex differences in circulating androgens. While androgens were detected in liver during late tadpole stages (stage 56-66), no sex differences were apparent. Androgens can be detected in tadpole tissues as early as tadpole stage 47. Δ5-3β-Hydroxysteroid dehydrogenase (HSD) histochemistry indicates that tadpole gonads and interrenals (and, to a much lesser extent, kidneys) are the only tissues capable of steroid biosynthesis. Interrenal HSD activity was present throughout tadpole development; in gonads, activity began before sexual differentiation. Thin-layer chromatographic separation of steroid metabolits following in vitro incubation with radioactive precursors indicated that the interrenals were more active than the gonads in tadpoles but not in juveniles. Thus, interrenals and gonads of developing X. laevis can synthesize Δ4 steroids (which include androgens) and, from early tadpole stages, are capable of metabolizing pregnenolone and testosterone, early and late compounds in the steroid synthetic pathway, respectively. The absence of dramatic sex differences in T or DHT levels during late tadpole and early juvenile stages suggests that while necessary, androgen secretion is not sufficient for masculinization of vocal neuroeffectors.