Reproduction in the Mexican leaf frog, Pachymedusa dacnicolor. VI. Presence and distribution of multiple GnRH forms in the brain.

The presence and distribution of gonadotropin-releasing hormone (GnRH) has been investigated in the Mexican leaf frog, Pachymedusa dacnicolor, brain during development and in the adult. The ontogenetic pattern of GnRH neurons illustrates their extracranial as well as intracranial sites. Immunohistochemical analysis indicates that GnRH-immunoreactive neurons appear during the metamorphic climax. They are located in the mesencephalon and subsequently other GnRH neurons appear in the peripheral terminal nerve and anterior preoptic area of the brain. Use of specific antisera and homologous combined with heterologous preabsorption tests indicate that mammalian and chicken GnRH-II-like peptide-containing neurons are differentially located within the brain, the former in the anterior preoptic area and peripheral terminal nerve and the latter in the mesencephalon. HPLC and RIA data suggest the presence of three forms of immunoreactive GnRH in the P. dacnicolor brain. A mammalian GnRH-like molecule and a chicken GnRH-II-like molecule are present. A third form, suspected to be [hydroxyproline9]mGnRH elutes before the mammalian GnRH.

Reproduction in the Mexican leaf frog, Pachymedusa dacnicolor. V. Immunohistochemical localization of gonadotropin-releasing hormone in the brain.

The presence and distribution of immunoreactive gonadotropin-releasing hormone (GnRH) in brains of adult male and female Pachymedusa dacnicolor has been studied immunohistochemically using antisera against mammalian, chicken-II, and salmon GnRHs. The distribution map of the immunoreactive-GnRH elements in the brain of P. dacnicolor is extremely simple, being limited to the anterior preoptic area-infundibulum-median eminence circuit. No sex- or reproductive status-related difference in either the distribution pattern or intensity of immunoreaction was revealed in this study. This is also the first immunohistochemical evidence of the presence of different structural forms of GnRH in the brain of an amphibian.

Reproduction in the Mexican leaf frog, Pachymedusa dacnicolor. III. The female.

The female of the Mexican leaf frog, Pachymedusa dacnicolor, displays a marked annual ovarian cycle. This consists of a long period of vitellogenic stasis, starting in early fall, soon after the breeding season, and ending in early spring. Oogonial proliferation continues throughout the year and new previtellogenic follicles are formed continuously. During the period of vitellogenesis, from spring to early summer, early, advanced, and postvitellogenic follicles are all found together, a situation that continues through the breeding season. This is correlated with the fact that a breeding female can lay three or more clutches per season. Breeding can begin as early as June and end as late as early September, with peak spawning activity taking place during July and August. An examination of the ovarian hormone secretion pattern in P. dacnicolor during the year revealed that plasma levels of testosterone and estradiol correlated with ovarian growth and attained highest levels in amplectant and ovulating females. Both hormones showed quite similar plasma levels and patterns of change during the annual reproductive cycle. Lowest plasma levels of testosterone and estradiol were found during fall and winter, in females possessing exclusively previtellogenic ovarian follicles. Plasma progesterone levels were maintained at a very low level throughout the year, except for the ovulatory surge, when amplectant and ovulating females may show a three- to sixfold increase. Plasma androstenedione showed a low peak during this phase of the reproductive cycle. Plasma levels of 5 alpha-dihydrotestosterone were 13 to 30 times lower than plasma testosterone levels. The potential roles of these gonadal steroids in controlling ovarian activity and reproduction are discussed briefly.

Reproduction in the Mexican leaf frog, Pachymedusa dacnicolor. II. The male.

The Mexican leaf frog, Pachymedusa dacnicolor, an inhabitant of the semiarid, subtropical Mexican lowlands, displays a well-defined seasonal testicular cycle. Testis weight seems to be a reliable index of the reproductive status of the animal and plasma levels of androgens (testosterone, T; 5 alpha-dihydrotestosterone, DHT; androstenedione, A) correlate not only with testicular growth, but with callosity development, reproductive behavior, and breeding. During the fall and winter, testis weight reaches its minimum as do plasma concentrations of T, DHT, A, and estradiol-17 beta (E). Plasma levels of progesterone (P) are maintained at a very low level throughout the year. During the fall and winter, spermatogenesis is almost entirely absent and the callosities are white and smooth. No signs of reproductive behavior (calling and amplexus) are evident. Late spring marks the initiation of spermatogenesis, testis weight increase, darkening of callosities, and increase in plasma levels of androgens. The magnitude of callosity development and the onset of calling and amplectant behavior are correlated with a great rise in plasma androgen levels. Although plasma T concentrations were higher than plasma levels of DHT and A, this androgen failed to stimulate the development of callosities and calling behavior in successfully castrated males.