The possible contribution of pituitary hormones to the heterochronic development of gonads and external morphology in overwintered larvae of Hynobius retardatus.

In Hynobius retardatus, most larvae in regions of low elevation metamorphose by autumn of the same year. However, larvae of some populations found in cold, mountainous ponds cannot metamorphose within the year and become aged, overwintered larvae. Gonadal development in larvae under the age of 1 year (larvae developed from eggs spawned in the same year) and in aged, overwintered larvae (spawned and hatched in previous years) was examined at the same developmental stage (stage 63, full-grown larval stage). The number of germ cells and the cross-sectional areas of the gonads were much larger in 2-season-overwintered (third year) larvae than in larvae under the age of 1 year. To obtain reliable probes for investigating the possible contribution of TSH, FSH and LH to metamorphosis and gonadal development, cDNAs for Hynobius TSHbeta, FSHbeta and LHbeta genes were cloned. Their expressions were analyzed by means of semi-quantitative RT-PCR in larvae under the age of 1 year and in 2-season-overwintered larvae. No differences were observed in expression levels of either TSHbeta or LHbeta between larvae under the age of 1 year and the overwintered larvae. In contrast, expression of FSHbeta was much higher in the overwintered larvae than in larvae under the age of 1 year. These results suggest that gonadal development proceeds gradually with age even in the overwintered larvae, but that metamorphosis is retarded, probably due to the larvae's cold habitat. Heterochronic development of gonads and external morphology has been demonstrated in H. retardatus, suggesting a potency for neotenic reproduction in this species.