The breeding season duration hypothesis: acute handling stress and total plasma concentrations of corticosterone and androgens in male and female striped plateau lizards (Sceloporus virgatus).

Acute glucocorticoid elevations can be adaptations to short-term stressors. The breeding season hypothesis predicts reduced glucocorticoid responsiveness to acute stressors in populations or species with short breeding seasons. The striped plateau lizard (Sceloporus virgatus) has a short breeding season in Arizona. We measured plasma corticosterone and total androgen levels (dihydrotestosterone and testosterone) following one of the four stress-handling treatments (0, 10, 60, or 180 min). In both sexes, longer handling stress yielded higher corticosterone; females had higher corticosterone than males at all time points. Androgens did not vary with handling duration, in either sex. Combining treatments, plasma androgens correlated positively with corticosterone (CORT) in females but not in males; plasma CORT and body mass residuals were negatively correlated in both sexes, suggesting lizards in poor body condition and/or not investing heavily in reproduction (follicle mass) have higher acute corticosterone. Total plasma androgens and body mass residuals were positively associated in males, but showed no association in females. The maximal CORT elevation after handling stress in this single-clutching species was of comparable magnitude to responses in related multi-clutching lizard species with longer breeding seasons. Using data from studies of multiple populations of three Sceloporus species, we found no relationship between the relative magnitude of the CORT increase and either latitude or elevation, two variables in the literature correlated with duration of the breeding season, and only weak relationships with geographic elevation and actual (not relative) stress-elevated CORT values in this multi-population comparison.

Fighting from the right side of the brain: left visual field preference during aggression in free-ranging male tree lizards (Urosaurus ornatus).

In a field study on male tree lizards (Urosaurus ornatus), we tested the hypothesis that aggression is lateralized, and predicted that resident males should preferentially use the left-visual field (LVF)/right hemisphere in aggressive interactions with other males. This prediction was based on results for a small but taxonomically diverse number of vertebrates indicating a left-eye preference/ right hemisphere specialization for aggression. Many lizards use postural displays to signal aggression and lateral eye placement prevents binocular vision of such displays. The fullshow is an aggressive display that is typically performed with the body held perpendicular to that of the opponent. We staged male-male encounters by introducing stimulus males to free-ranging males (hereafter called focal males), and then recorded both the focal male's behavior and the visual field that he used to view the intruder while performing fullshow displays. When intruders were introduced in the right visual field (RVF), focal males were equally likely to use either the LVF or RVF to perform their first fullshow. However, in these RVF introductions focal males were more likely to turn and use the LVF if the intruder was smaller. In contrast to RVF introductions, in LVF introductions focal males were significantly more likely than expected by chance to continue to use the LVF to view the intruder when performing the first fullshow. Furthermore, if focal males performed their first fullshow with the intruder in their LVF, they subsequently gave more aggressive displays than focal males that performed their first display with the intruder in their RVF. Additionally, charge (a highly aggressive behavior that usually leads to direct physical contact and a bite) was significantly more likely to occur while the LVF was being used to view the opponent. These results are consistent with the hypothesis that aggression is lateralized to the right hemisphere. This study is one of only a few examples of laterality of brain function demonstrated in a free-ranging vertebrate.

Pollination, breeding system, and genetic structure in two sympatric Delphinium (Ranunculaceae) species.

Two sympatric Delphinium species, D. barbeyi and D. nuttallianum, are ecologically and morphologically similar. However, D. barbeyi has multiple, large inflorescences while D. nuttallianum has a single, small inflorescence. These differences in floral display should result in greater intraplant pollen transfer in D. barbeyi, leading to higher rates of self-pollination through geitonogamy. Reduced gene flow by pollen should in turn produce greater population differentiation among populations of D. barbeyi relative to D. nuttallianum. We tested these predictions by comparing pollinator behavior, breeding systems, outcrossing rates, and population genetic structure of sympatric populations of the two species in Colorado. Bumble bee and hummingbird pollinators visit more flowers and inflorescences per foraging bout in D. barbeyi than in D. nuttallianum. The species differed in breeding system; D. barbeyi produced more seeds by autogamy (9 vs. 2%) than D. nuttallianum and suffered no reduction in seed set in hand-self vs. outcross pollinations, in contrast to a 41% decline in D. nuttallianum. The outcrossing rate in one D. barbeyi population was 55%, but ranged from 87 to 97% in four D. nuttallianum populations. Genetic differentiation among population subdivisions estimated by hierarchical F statistics was >10 times greater in D. barbeyi ( = 0.055-0.126) than D. nuttallianum ( = 0.004-0.009) at spatial scales ranging from metres to 3.5 km. Spatial autocorrelation analysis also indicated more pronounced local genetic structure in D. barbeyi than D. nuttallianum populations. Fixation indices (F(IS)) of D. barbeyi adults were much lower than expected based on mating system equilibrium and suggest that differences in the degree of self-compatibility and/or the timing of postpollination selection/inbreeding depression between the two species further contribute to the genetic differences between them.

Signals and behavioural responses are not coupled in males: aggression affected by replacement of an evolutionarily lost colour signal.

Male Sceloporus virgatus lack the blue abdominal patches which are used during aggressive encounters in other Sceloporus lizards. Herein we report that, despite having lost this signal, males have retained a behavioural response to experimentally restored blue abdominal patches. We tested two adaptive hypotheses: selection acted primarily upon signallers or selection acted upon both signallers and receivers. The first predicts that only the signal is lost and that male interactions should be affected by the restoration of blue patches. The latter predicts that both the signal and behavioural response are lost and the display of the restored blue patches should have no effect on male-male interactions. We compared the behaviour of receivers in paired encounters where one male (signaller) had blue-painted abdominal patches to a set of trials where both males had white-painted abdomens, unmanipulated abdomens or a novel-painted pattern. The receivers of the blue-painted signal were more likely to display submissive behaviour. The receivers in either the unmanipulated, white-painted or novel-painted signal trials were more likely to display neutral behaviour. These results support the hypothesis that receivers have retained a behavioural response and selection has acted primarily on the signaller. We believe this is the first documentation of males responding to an evolutionarily lost signal in conspecific males.

A critical period for the organization of alternative male phenotypes of tree lizards by exogenous testosterone?

Male tree lizards (Urosaurus ornatus) exhibit permanent adult differences in color and size that are functionally linked with behavioral differences: males with a central blue throat patch are territorial, more aggressive, and smaller than males lacking the patch, who are nomads or satellites foregoing territory defense. Gonadectomy or long-lasting hormone implants in hatchlings affect the development of these permanent adult differences, but similar manipulations in adults are ineffective. Analogous early hormone actions in vertebrate sexual differentiation typically must occur during a critical period. Whether there is a critical period for early hormone actions affecting development of male types in tree lizards is unclear. We followed a protocol identical to our previous experiments on hatchlings, but we manipulated testosterone (T) in later-aged juveniles to determine if they remained sensitive to such hormone manipulations. Testosterone implants given at 60 days posthatching (day 60) did not alter adult throat color, indicating that sensitivity of throat color to T declines by day 60. This contrasts to our earlier work where adult color was affected by T manipulations on day 30. However, size and throat color responded differently to exogenous T, as juveniles given T implants at day 60 grew less than control-implanted males.

Influence of androgens on differentiation of secondary sex characters in tree lizards, Urosaurus ornatus.

Vertebrates species vary in the degree to which the sexes differ in their expression of secondary sex characters, which can be expressed in one sex but not the other, fully expressed in both sexes, or expressed to different degrees in the two sexes. Sex steroid hormones contribute to the development of sex differences, either through action early in life (organization), following sexual maturation (activation), or both. However, relatively little is known about the contributions of sex steroid hormones to species-level variation in sexual dimorphism. We began to address this by assessing in tree lizards, Urosaurus ornatus, the effects of testosterone (T) and dihydrotestosterone (DHT) on expression of the male-typical traits: femoral pore secretions, accessory sex tissues, patches of ventrolateral blue, intensity and hue of throat color, and body size. We administered long-lasting hormone implants of these androgens to intact or ovariectomized adult females, intact hatchling females, and intact hatchling males to determine the relative contribution of organizational and activational influences of androgens on sexual differentiation of these traits. Waxy femoral pore secretions and full intensity of the orange background color of the throat fan (dewlap) required only activation and both androgens were equally effective. Both androgens caused hypertrophy of accessory tissues associated with the hemipenes, but only in hatchling males. Full expression of ventrolateral blue patches required organization by DHT. Androgens had complex organizational actions on growth. In the doses used in the experiment, DHT inhibited body-length growth but not mass growth of hatchling males. Only T inhibited the growth (length and mass) of hatchling females. Because earlier work found that castration also reduced the growth of hatchling males, the organizational effects of androgen on growth might be dose-related. Results for hormonal control of dewlap type were complex. In this population dewlap color is solid orange in females, but polymorphic in males with 50% expressing solid orange (O) and 50% orange-blue (orange with a central blue patch, OB). These color morphs represent permanent male alternatives: OB males are territorial and more aggressive than O males who do not defend territories. Results of the hormone manipulations are most consistent with the interpretation that dewlap morph type requires both organization and activation by androgen. T appears to be more important in organizing morph differences, whereas DHT appears to be more important in activating expression of underlying morph differences. Females only expressed male-typical OB dewlaps when given DHT, and only about 50% did so.

Early exposure to androgens affects adult expression of alternative male types in tree lizards.

Males of many species exhibit strongly dimorphic reproductive behavior and morphology associated with alternative reproductive tactics. Little is known about the physiological control of these individual differences. The relative plasticity hypothesis proposes that such within-sex differences arise from either organizational or activational actions of sex steroid hormones depending on whether adults can switch tactics or not. This hypothesis predicts that differences between individuals in a species where adults cannot switch between tactics (are "fixed") should be organized by early actions of hormones. Three experiments explored the possible organizational role of testosterone (T) on the development of male alternatives in the tree lizard (Urosaurus ornatus), which has two fixed male morphs. Orange (O) males have a mostly orange throat fan (dewlap), are nonterritorial nomads, and are larger than orange-blue (OB) males which have an orange dewlap with a central blue patch and are territorial. In the first experiment intact males on the day of hatching were (1) sham-operated and implanted with empty capsules, (2) castrated, or (3) testosterone-implanted. As adults, the castration group had more O males than the control and the T-implanted group had more OB males than the control. Adult body size in castrated and T-implanted groups parallelled naturally occurring morph differences, but both were smaller than controls. A second experiment with a lower dose yielded similar results for dewlap type and growth. In a third experiment, intact males were given an empty capsule or a T-implant at 30 days posthatching. Again, the frequency of OB males in the T-implanted group was significantly greater than that in the empty implant group, indicating that either the critical period extends past Day 30 or there is no well-defined critical period. Together, these results support the hypothesis that the organizational action of T or one of its metabolites contributes to the differentiation of these within-sex differences.

An investigation of the alarm response in Bufo boreas and Rana cascadae tadpoles.

Tadpoles of the western toad (Bufo boreas) and of the Cascades frog (Rana cascadae) show an alarm reaction to an extract containing chemical cues from damaged conspecifics. The mean time spent by individual B. boreas tadpoles in the half of the test tank to which the extract solution was added was significantly lower than expected by chance. Activity was also significantly greater in Bufo extract tests than in control tests. Tadpoles did not avoid an extract of another tadpole species (Hyla regilla). Rana cascadae tadpoles did not avoid areas containing Rana extract but did significantly increase their level of activity in response to the extract. These results suggest that the R. cascadae tadpole alarm reaction exists but differs from the B. boreas reaction.