Androgen and gonadotropin effects on male mate calls in South African clawed frogs, Xenopus laevis

Mate calling is a prominent reproductive behavior of male South African clawed frogs. Calls consist of alternating slow- and fast-amplitude-modulated trills. Each trill is made up of a series of clicks. The effects of administration of exogenous gonadotropin and androgen on mate calling were studied in male Xenopus laevis. Males were paired with unreceptive female frogs to elicit maximal calling. The amount of time each animal spent calling during the testing period, the peak fundamental frequency of the calls, the rate of calling, and the interclick interval (ICI, a measure of the temporal patterning of the calls) were measured in intact, castrated, and hormone-replaced frogs. Injection of human chorionic gonadotropin (HCG) into intact frogs increased the amount of time spent calling and the ICI relative to measures taken after water injection. Castrated males did not call even when given HCG. Testosterone and dihydrotestosterone treatment reinstated calling in castrates and increased circulating levels of androgens. When androgen-replaced castrated males were injected with HCG, the amount of time spent calling increased and approached levels of intact, HCG-injected males. The above results suggest that androgens are necessary for the production of calls. Gonadotropins appear to play an important role in mate calling, a role at least partly independent of effects on testicular androgen synthesis.     

Androgen directs sexual differentiation of laryngeal innervation in developing Xenopus laevis

In adult Xenopus laevis, innervation of the vocal organ is more robust in males than in females. This sex difference originates during tadpole development; at stage 56, when the gonads first differentiate, the number of axons entering the larynx is the same in the sexes, but by stage 62, innervation is greater in males. To determine if androgen secretion establishes sex differences in axon number, we treated tadpoles with antiandrogen or androgen beginning at stage 48 or 54 and counted laryngeal nerve axons at stage 62 using electron microscopy. When male tadpoles were treated with the antiandrogen hydroxyflutamide, axon numbers were reduced to female-typical values; axon numbers in females were unaffected by antiandrogen treatment. When female tadpoles were treated with the androgen DHT (dihydrotestosterone), axon numbers were increased to male-like values. These findings suggest that endogenous androgen secretion during late tadpole stages in males is required for the sexual differentiation of laryngeal innervation observed from stage 62 on. Because androgen treatment and laryngeal innervation affect myogenesis in postmetamorphic frogs, numbers of laryngeal dilator muscle fibers were determined for hormonally manipulated tadpoles. At stage 62, vehicle-treated males had more laryngeal axons than females; laryngeal muscle fiber numbers did not, however, differ in the sexes. Both male and female tadpoles, treated from stage 54 with DHT, had more muscle fibers at stage 62 than vehicle-treated controls. Thus, while endogenous androgen secretion during late tadpole stages is subthreshold for the establishment of masculinized muscle fiber numbers, laryngeal myogenesis is androgen sensitive at this time and can be increased by suprathreshold provision of exogenous DHT. A subgroup of tadpoles, DHT treated from stage 54 to 62, was allowed to survive, untreated, until postmetamorphic stage 2 (PM2: 5 months after metamorphosis is complete). Androgen treatment between tadpole stages 54 and 62 does not prevent the ontogenetic decrease in axon numbers characteristic of laryngeal development. In addition, the elevation in stage 62 axon numbers produced by DHT-treatment at late tadpole stages was not associated with elevated numbers of laryngeal muscle fibers at PM2. Juvenile males normally maintain elevated axon numbers (relative to final adult values) through PM2 and the presence of these additional axons may result from-rather than contribute directly to—laryngeal muscle fiber addition. 1994 John Wiley & Sons, Inc.     

Phorbol 12-myristate 13-acetate stimulates proliferation and ductal morphogenesis and inhibits functional differentiation of normal rat mammary epithelial cells in primary culture

The effect of the tumor promoter phorbol 12-myristate 13-acetate (PMA) on proliferation and differentiation of normal mammary epithelial cells from 50-day-old virgin rats was investigated using a model system that allows for full morphological and functional development of the cells. In this model, mammary epithelial cells are grown within a reconstituted basement membrane in a defined serum-free medium. PMA at a concentration of 10^?6 M effected translocation of protein kinase C from cytosol to membrane. At the same concentration, it stimulated cell proliferation both in the presence and absence of EGF, and this stimulation was observed even when PMA exposure was limited to 15 min at the time of each media change. In contrast to the effect on proliferation, PMA at concentrations of 10^?7 and 10^?6 M inhibited functional differentiation as assessed by casein accumulation. Phorbol 12,13-dibutyrate at 10^?6 M also stimulated proliferation and inhibited casein accumulation and was more effective than PMA in both cases. In contrast, the nonactive tumor promoter 4-α PMA had no effect on either proliferation or differentiation. One of the most striking effects of PMA was its ability to stimulate an atypical ductal morphogenesis, as manifested by the formation of intricate web-like colonies, and to inhibit the development of the well-differentiated alveolar-like multilobular colonies. PMA was also shown to completely suppress the growth of the squamous-like colonies that develop when EGF is absent or deficient. These effects of phorbol esters in mammary epithelial cells to stimulate proliferation, inhibit functional differentiation, and stimulate the development of ductal colonies are consistent with the suggestion that the signal transduction pathways evoked by PMA could act to stimulate the growth of initiated cells or render normal cells more sensitive to carcinogen. © 1994 Wiley-Liss, Inc.     

Infection of the athymic rat by Toxoplasma gondii

In contrast to normal rats which are resistant to T. gondii infection (10(7) tachyzo?tes), athymic rats did not survive an intraperitoneal infection with 10(3) toxoplasma. When nude rats were injected intravenously with lymph node cells from hirsute littermates, they became resistant in a dose-dependent manner to the infection. In addition, reconstituted athymic rats having survived for more than 4 months the first infection were also protected against a second challenge with 10(5) tachyzo?tes. Anti-T. gondii antibody levels detected in reconstituted athymic rats were related to protection. These preliminary findings suggest that T-dependent immunity is essential in the development of effector mechanisms involving antibodies in resistance to toxoplasmosis.     

Prostaglandin E2 induces receptive behaviors in female Xenopus laevis

The object of this study was to examine the effects of exogenous and endogenous prostaglandin E2 (PGE2) on the sexual behavior of female South African clawed frogs, Xenopus laevis. Ticking and leg extension, which communicate sexual unreceptivity to males, were studied in intact, ovariectomized, and ovariectomized-oviductectomized females. The onset of the PGE2 behavioral effect occurs within 30 sec to 3 min of injection for intact and ovariectomized females; for ovariectomized-oviductectomized females, the latency period for the effect ranges from 10-20 min. PGE2 induced receptivity in doses as low as 0.03 microgram/frog. Injection of the prostaglandin synthesis inhibitors, indomethacin and flurbiprofen (FBP), blocked chorionic gonadotropin- (HCG-) induced behavioral receptivity, suggesting that endogenous prostaglandin synthesis may have a role in regulating female sexual behavior. Flurbiprofen blockade of HCG-induced receptivity was reversed by PGE2 administration, suggesting that FBP's effects are PG synthesis-specific.