The vocal motor neurons of Xenopus laevis: development of sex differences in axon number.

Sex differences in the number of muscle fibers in the larynx of clawed frogs (Xenopus laevis) develop after metamorphosis. In order to examine possible contributions of neural innervation to this process, we prepared sections of the laryngeal nerve from tadpole stage 56, when the sexes can first be distinguished, through adulthood, and counted axons on electron micrographs. The adult number of axons is achieved by a sexually differentiated pattern of axonal addition and loss. Axon numbers are high at tadpole stage 56 and equal for males and females; thereafter, males have more axons. Sex differences are most pronounced at tadpole stage 62 because between stages 59 and 62 the number of laryngeal axons in males increases by an average of 119 per nerve. Ultrastructural evidence is congruent with the hypothesis that new axons are added to the laryngeal nerve between tadpole stages 56 and 62. The loss of axons from the laryngeal nerve is greater for females than for males. Between tadpole stages 56 and adulthood, overall axon number decreases by 47% in males and by 64% in females. Signs of axonal degeneration are present in both sexes before metamorphosis but are rare at juvenile or adult stages. The numbers of axons in juvenile frogs do not differ from those in adults and continue to be greater in males than in females. In contrast to the amount of axon addition and loss, the timing of axon loss and the percentage of myelinated axons is the same for males and females throughout development. Thus sex differences in the innervation of laryngeal muscle originate before metamorphosis and could contribute to the marked sex differences in muscle fiber addition that occur thereafter.