Identification and properties of steroid-binding proteins in nesting Chelonia mydas plasma.

We report for the first time the presence of a sex steroid-binding protein in the plasma of green sea turtles Chelonia mydas, which provides an insight into reproductive status. A high affinity, low capacity sex hormone steroid-binding protein was identified in nesting C. mydas and its thermal profile was established. In nesting C. mydas testosterone and oestradiol bind at 4 degrees C with high affinity (K (a) = 1.49 +/- 0.09 x 10(9) M(-1); 0.17 +/- 0.02 x 10(7) M(-1)) and low binding capacity (B (max) = 3.24 +/- 0.84 x 10(-5) M; 0.33 +/- 0.06 x 10(-4) M). The binding affinity and capacity of testosterone at 23 and 36 degrees C, respectively were similar to those determined at 4 degrees C. However, oestradiol showed no binding activity at 36 degrees C. With competition studies we showed that oestradiol and oestrone do not compete for binding sites. Furthermore, in nesting C. mydas plasma no high-affinity binding was observed for adrenocortical steroids (cortisol and corticosterone) and progesterone. Our results indicate that in nesting C. mydas plasma temperature has a minimal effect on the high-affinity binding of testosterone to sex steroid-binding protein, however, the high affinity binding of oestradiol to sex steroid-binding protein is abolished at a hypothetically high (36 degrees C) sea/ambient/body temperature. This suggests that at high core body temperatures most of the oestradiol becomes biologically available to the tissues rather than remaining bound to a high-affinity carrier.

Patterns of lipid storage and mobilisation in the female green sea turtle ( Chelonia mydas).

Reproductive data from southern Queensland indicate that vitellogenesis in female Chelonia mydas takes approximately 8 months and is followed by a migration to a breeding area. At Heron Island, females lay multiple clutches over approximately 3 months. To investigate how females mobilise and store lipid during the breeding season we collected plasma, yolk, and fat tissue samples from females at a variety of stages during the nesting season. In breeding females, concentrations of plasma triglyceride increased seasonally. They reached peak concentrations during vitellogenesis and courtship, remained high throughout the nesting season, and then declined to a nadir after the last clutch. Plasma protein concentration increased throughout the breeding season, peaking following the last clutch for the season. Yolk lipids were highest during courtship and were similar throughout the nesting season, suggesting that uptake of lipid by ovarian follicles is completed prior to the beginning of the nesting season. Plasma triglyceride decreases in females with prolonged periods of unsuccessful nesting, and total lipid levels in adipose tissue and follicle yolks were significantly lower in atretic females. It appears that: (1) endogenous energy reserves can be reduced by stochastic environmental events (such as those reducing nesting success), and (2) a metabolic shift signalling the end of the nesting season is characterised by a drop in plasma triglycerides and slight increase in total plasma protein.

Lack of consistent hormonal responses to capture during the breeding season of the bearded dragon, Pogona barbata.

The bearded dragon (Pogona barbata: Agamidae) is a diurnal, oviparous, multi-clutching lizard from Australia. We examined plasma hormonal responses to capture in males and females during the spring breeding season. Corticosterone concentrations at capture (0 h; < or =3 min after capture) were low (males: 1.81+/-0.63 ng/ml; females 2. 23+/-0.47 ng/ml) and within sexes were unrelated to the time of the day, snout-vent length or, in females, reproductive condition (vitellogenic, gravid, assumed spent). Corticosterone concentrations at capture were significantly and inversely correlated with body condition in males, but not in females. Unexpectedly, neither sex showed significant changes in mean concentrations of corticosterone at 3.5 or 24 h after capture compared with 0 h values. Corticosterone concentrations at 3.5 h after capture did not differ between dragons bled at capture or not. Concentrations of progesterone in both the sexes did not change between 0 h and 3.5 or 24 h after capture. Testosterone concentrations in males at capture were moderate (10.1+/-2.2 ng/ml), and unchanged at 3.5 h after capture. The adrenocortical axis of adult bearded dragons in the breeding season seems remarkably unresponsive to capture compared with many other reptiles. Low adrenocortical responses to capture may be a feature of reptiles known to adjust well to captivity.

Seasonal patterns of plasma steroid hormones in males and females of the bearded dragon lizard, Pogona barbata.

Pogona barbata is an Australian lizard that produces several large clutches of eggs between August and December (spring to early summer). Mating takes place around ovulation. The seasonal pattern of reproductive hormones in males and females of P. barbata was determined by radioimmunoassay of plasma progesterone (P), estradiol-17beta (E-17beta), corticosterone (B), and total androgen (TA). In females, P began to rise in August and was elevated from September to December. Corticosterone and TA were detectable but low and did not vary with time of year or reproductive condition. Estradiol-17beta was only detectable in a few females and exhibited no elevation with vitellogenic activity. These results suggest that B and TA are not involved in female reproduction. Estrogens may be either so low they could not be detected or they were present in a form other than estradiol-17beta. The high sensitivity of the estradiol-17beta radioimmunoassay suggests the latter. In males, TA peaked at the beginning of spring. They then declined to a minimum during November and December. However, concentrations recovered in the postbreeding activity period, January to April (summer and autumn). These patterns are consistent with the observation of maximum spermatogenic activity in spring, followed by the cessation of spermatogenesis directly after the breeding period and testicular recrudescence in February (late summer).

Interactions between behavior and plasma steroids within the scramble mating system of the promiscuous green turtle, Chelonia mydas.

We measured plasma androgen (combined testosterone and 5alpha-dihydrotestosterone) (A) and corticosterone (B) in the promiscuous green turtle (Chelonia mydas) during courtship in the southern Great Barrier Reef. This study examined if reproductive behaviors and intermale aggression induced behavioral androgen and adrenocortical responses in reproductively active male and female green turtles. Associations between reproductive behavior and plasma steroids were investigated in green turtles across the population and within individuals. Levels across a range of both asocial and social behaviors were compared including (a) free swimming behavior; (b) initial courtship interactions; (c) mounted behavior (male and female turtles involved in copulatory activities); (d) intermale aggression (rival males that physically competed with another male turtle or mounted males recipient to these aggressive interactions); and (e) extensive courtship damage (male turtles that had accumulated excessive courtship damage from rival males). Behavioral androgen responses were detected in male turtles, in that plasma A was observed to increase with both attendant and mounted behavior. Male turtles who had been subjected to intermale aggression or who had accumulated severe courtship damage exhibited significantly lower plasma A than their respective controls. No pronounced adrenocortical response was observed after either intermale aggression or accumulation of extensive courtship damage. Female turtles exhibited a significant increase in plasma B during swimming versus mounted behavior, but no change in plasma A. We discuss our results in terms of how scramble polygamy might influence behavioral androgen interactions differently from more typical combative and territorial forms of male polygamy.

Plasma steroid hormone profiles and reproductive biology of the epaulette shark, Hemiscyllium ocellatum.

Examination of the reproductive biology of the oviparous epaulette shark, Hemiscyllium ocellatum, was conducted on a wild population. Male sharks were found to reach maturity at between 55-60 cm total length (TL) and female sharks mature around 55 cm TL. Blood samples collected from mature male and female sharks were analyzed for sex steroid hormones to examine seasonal hormone patterns. Plasma samples were analyzed via radioimmunoassay techniques with female samples measured for estradiol, progesterone, and androgen concentrations, and male samples measured for androgen concentrations. Male androgen concentrations showed a single broad peak from July to October with maximum hormone concentrations (60 ng/ml) occurring in August. Male androgen concentrations were lowest in December-February (<20 ng/ml), and appeared to correlate with reproductive activity and water temperature. Female androgen concentrations were an order of magnitude lower than those for males and showed peaks in June (6 ng/ml) and December (8 ng/ml). Estradiol concentrations in females peaked during the months of September-November (0.5 ng/ml) coinciding with the egg laying period. Progesterone concentrations ranged up to 0.5 ng/ml prior to the mating season. Observations of ova size and egg production showed eggs develop in pairs and ova are ovulated at a size of 25-27 mm. Females lay eggs from August to January. Males were observed with swollen claspers from July through December, with the highest amount of sperm storage in the epididymis occurring between August through November. Our observations indicate that epaulette sharks in the waters near Heron Island mate from July through December. J. Exp. Zool. 284:586-594, 1999.

Plasma steroid interactions during high-density green turtle nesting and associated disturbance.

Raine Island in the Northern Great Barrier Reef constitutes an extremely high-density green turtle (Chelonia mydas) rookery. On this island, competitive interactions for nesting space and subsequent disturbance of individual nesting are widespread. High-density nesting often delays successful oviposition by one or more nights. There is little information on how hormones in female reptiles interact during competitive reproductive events in such high-density nesting populations. In this three-part study we investigated the interactions between density (within and between rookery/ies), nesting success and failure, and plasma steroid profiles in green turtles. First, we compared levels of plasma corticosterone (B) and combined testosterone + 5alpha-dihydrotestosterone (T + DHT) in turtles during five stages of oviposition in both a high-nesting-density sector (1 turtle/m2) and a low-nesting-density sector (0.1 turtle/m2). Second, we investigated the relationship between increasing delays (0, 1, 2, 3, and 6 days) in successful oviposition and the plasma steroids B and T + DHT. Third, we assessed a comparative measure of steroid hormone levels of females at low-density sites on Raine Island (high-density rookery) and Number Seven Sandbank (low-density rookery). Despite a significant trend suggesting high-density nesting turtles elaborated more plasma B than turtles in low-density sectors, the magnitude of this increase was small. We suggest that this increase may be an artifact of increased metabolic demand and hence catabolism of energy substrates associated with high-density nesting. Plasma T + DHT remained stable in response to density-dependent effects associated with nesting. Furthermore, prolonging successful oviposition because of multiple nightly disturbance failed to elicit any change in either plasma B or T + DHT. These data suggest that green turtles may be exhibiting adrenal desensitization to prevent both physical and behavioral disturbances interfering with reproduction. We suspect that down-regulating the acute adrenocortical response may represent an adaptive trade-off mechanism for optimizing current reproductive success at the potential expense of survivorship.

Plasma steroid profiles in nesting loggerhead turtles (Caretta caretta) in Queensland, Australia: relationship to nesting episode and season.

Plasma levels of four hormones-progesterone (P), testosterone (T), estradiol 17-beta (E2), and corticosterone (B)-were measured in samples taken from nesting female loggerhead turtles (Caretta caretta) by using specific radioimmunoassays. Samples were taken in an early, middle, or late period during the summer nesting season from females at first, second, third, or > fourth nesting episodes, defined as successive within-season nesting events, at Mon Repos Beach, Queensland, Australia. Data on individual patterns of nesting, collected over the past 20 years by the Queensland Turtle Research Project, and the seasonal nesting data, were analyzed with respect to influences on hormonal profiles. Circulating levels of E2 were mostly undetectable, suggesting either that this estrogen is not produced at this time of nesting, or that, perhaps, another estrogen may be present that is not detected by the specific radioimmunoassay. P, T, and B profiles in the nesting females were associated with the individual turtles' progression through successive nesting episodes, with a marked decline in all three hormones by the last (> 4) nesting episode of the season. Nesting episode accounted for significant changes that were related to season, in that nesting episode and season were significantly correlated. These patterns were observed in the population, when singly sampled at each time period or nesting episode, and in individual females sampled repeatedly over time. Moreover, T and B were highly and significantly correlated in females at all nesting episodes and time periods, and in the singly and repeatedly sampled females. The magnitude of change in T and B over time was also highly and significantly correlated in repeatedly sampled females. Together these results suggest the hypothesis that T and B interact over the period of successive nesting and may be involved in reproductive functions such as the mobilization of reserves for egg production in C. caretta.

Plasma triglyceride and beta-hydroxybutyric acid levels in red-sided garter snakes (Thamnophis sirtalis parietalis) at emergence from hibernation.

Measurement of plasma levels of triglycerides and beta-hydroxybutyric acid in females and males of the red-sided garter snake (Thamnophis sirtalis parietalis) suggest that the former may provide a useful physiological marker of condition. Levels of triglycerides at emergence from hibernation during a month of natural aphagia were significantly greater in females than in males and she-males, a subset of the male population that mimics females. Higher levels of triglycerides in the females may be attributed to their greater body mass per unit length, which was correlated with the level of triglycerides. Plasma triglyceride levels declined in females within one month of emergence, at the onset of feeding, and were unrelated to mating.

Prostaglandin F2 alpha, sexual behavior and ovarian estrogen synthesis in garter snakes (Thamnophis sirtalis parietalis).

1. High levels of prostaglandin F2 alpha (PGF2 alpha; 10-18 ng) were detected in copulatory material removed from the cloacae of female Thamnophis sirtalis parietalis immediately after mating. 2. In vitro, ovarian fragments treated with PGF2 alpha secreted significantly more estrogen than control fragments. 3. Together these results suggest that a post-mating surge in estrogen secretion that occurs in this species may be influenced by a substantial rise in PGF2 alpha that occurs after mating. 4. These findings are consistent with the hypothesis that post-mating elevations in plasma levels of PGF2 alpha in females are derived from male copulatory material.