Information out of the blue: phenotypic correlates of abdominal color patches in Sceloporus lizards.

Colorful ornaments are important visual signals for animal communication that can provide critical information about the quality of the signaler. In this study, we focused on different color characteristics of the abdominal patches of males of six lizard species from the genus Sceloporus. We addressed three main objectives. First, we examined if size, brightness, saturation, and conspicuousness of these ornaments are indicative of body size, condition, immune function, or levels of testosterone and corticosterone. Second, we evaluated if the distinct components of these abdominal patches (blue or green patches and black stripes) transmit similar information about the signaler, which would support the redundant signal hypothesis, or if these components are related to different phenotypic traits, which would support the multiple message hypothesis. Third, we compared the phenotypic correlates of these ornaments among our six species to understand the degree of conservatism in the signaling patterns or to find species-specific signals. Using data collected from males in natural conditions and a multi-model inference framework, we found that in most species the area of the patches and the brightness of the blue component are positively related to body size. Thus, these color characteristics are presumably indicative of the physical strength and competitive ability of males and these shared signals were likely inherited from a common ancestor. In half of the species, males in good body condition also exhibit relatively larger blue and black areas, suggesting that the expression of these ornaments is condition-dependent. Abdominal patches also provide information about immunocompetence of the males as indicated by different correlations between certain color characteristics and ectoparasite load, counts of heterophils, and the heterophil:lymphocyte ratio. Our findings reveal that area and brightness of the abdominal patches signal the size and body condition of males, whereas blue saturation and conspicuousness with respect to the surrounding substrate are indicative of immune condition, thus supporting the multiple message hypothesis. However, some of these correlations were not shared by all species and, hence, point to intriguing species-specific signals.

Steroid hormones, ectoparasites, and color: Sex, species, and seasonal differences in Sceloporus lizards.

Testosterone, in addition to promoting the expression of sexual ornaments can negatively affect immune function, leaving individuals more susceptible to parasites (immunocompetence handicap hypothesis). Immunosuppressive effects of testosterone also can occur indirectly, through increased glucocorticoid hormones (corticosterone, cortisol). Therefore, the expression of sexual ornaments and the ability to respond to parasites and diseases may be influenced by the interaction between testosterone and corticosterone. In this study we examined correlations of both testosterone and corticosterone with ectoparasite load and with expression of colorful sexual ornaments (patches on belly and throat) in three species of Sceloporus lizards (S. grammicus, S. megalepidurus, S. torquatus). In addition, we evaluated contributions of sex, body condition, and reproductive season. We expected that individuals with higher testosterone and lower corticosterone levels would have more colorful ornaments than individuals with higher corticosterone levels. In addition, if testosterone has negative effects on immune function but only at higher corticosterone levels, individuals with higher levels of testosterone and corticosterone should have higher ectoparasite loads. Contrary to these expectations, we did not detect an interaction between testosterone and corticosterone statistically affecting either ectoparasite load or the expression of colorful ornaments. Further, we did not find a positive association of either testosterone or corticosterone on ectoparasite loads in any of the three study species. Only in S. grammicus males was a hormone statistically associated with ectoparasite load, but it was a negative association with testosterone. The relationships between both hormones and different aspects of the colorful patches (brightness and chroma) varied drastically among the three species as well as between sexes. Hence, even among congeneric species, we did not observe consistent patterns among color, steroid hormones and ectoparasites. Different associations between these variables may reflect different physiological strategies for the production of colorful signals and immune defense.

Melanization, α-melanocyte stimulating hormone and steroid hormones in male western fence lizards from nine populations.

Hormones can mediate suites of correlated traits. Melanocortins regulate melanin synthesis and elements of the melanocortin system can directly, and indirectly, affect a number of other traits, such as stress reactivity. Trait correlations within the melanocortin system have been studied mainly in birds and mammals but less so in reptiles. We examined adult male western fence lizards (Sceloporus occidentalis) and if melanization was correlated with plasma levels of three hormones, including peptide hormone α-melanocyte stimulating hormone (α-MSH), testosterone and corticosterone, and ectoparasite loads. This lizard is darker at higher elevations in California, and we compared five high-elevation and four low-elevation populations during comparable periods of the breeding season at each site. We first validated use of an α-MSH assay kit with lizard plasma. Since Anolis carolinensis is one of the few species with published values for α-MSH plasma levels, we assayed both Anolis and Sceloporus plasma and compared hormone values to those we generated for Anolis to the publish values. We also evaluated effects of different methods of storing spiked plasma pools on resulting α-MSH concentrations. Plasma levels of α-MSH did not differ significantly, but some populations differed significantly in mean corticosterone and mean testosterone. Combining all individuals from the nine populations, we found that individual variation in α-MSH was not associated with individual variation in melanization, but levels of α-MSH were positively associated with plasma testosterone and negatively associated with corticosterone. The lack of association between individual levels of melanization and expression of most other traits differs from a growing number of within-population studies of melanization, and we discuss what differences in physiological mechanisms could produce different hypothetical patterns. Circulating levels of -MSH are only one element of the melanocortin system; in situ synthesis of α-MSH by the skin and the diversity of melanocortin receptors could also contribute to variation in traits mediated by the melanocortin system and should be examined.

Correlates of melanization in multiple high- and low-elevation populations of the lizard, Sceloporus occidentalis: Behavior, hormones, and parasites.

Hormones mediate the expression of suites of correlated traits and hence may act either to facilitate or constrain adaptive evolution. Selection on one trait within a hormone-mediated suite of traits may lead to a change in the strength of the hormone signal, causing changes in correlated traits. Growing evidence suggests that melanization, which is in part regulated by hormonal signals, is tightly linked to other traits, such as aggression and stress physiology. Here, we examine six populations of Sceloporus occidentalis lizards differing in degree of melanization (three dark higher-elevation populations and three less-melanized lower-elevation populations) to investigate potential correlations between behavior, hormones, and parasites. We measured aggression by recording behavioral responses of males to staged territorial intrusions; behavior was summarized by two principal components. Analysis revealed that males in the three darker populations signaled aggression less often and made more physical contact than males in the lighter populations. Analyses of plasma steroid hormones (corticosterone and testosterone) revealed significant population differences, but counter to expectation higher aggression was associated with lower testosterone compared across populations. Finally, the three darker populations had higher mean mite loads than the three lighter populations. Overall, this array of phenotypic correlations does not parallel patterns of within-population differences in melanization found in other vertebrates, suggesting that hormonal correlations do not constrain phenotype variation across populations in this species. Given this contradiction between population- and individual-level variation, we urge more study at both levels of variation in traits potentially associated with melanization in other vertebrates.

Populations of the Lizard, Sceloporus occidentalis, that Differ in Melanization have Different Rates of Wound Healing.

Mechanisms underlying production of animal coloration can affect key traits besides coloration. Melanin, and molecules regulating melanin, can directly and indirectly affect other phenotypic traits, such as immune function. We asked whether melanization and a whole-organism measure of immune function are associated with wound healing. Working with two populations of adult male western fence lizards, Sceloporus occidentalis, we compared one high-elevation and one low-elevation population in California where individuals are increasingly darker at higher elevations, measuring wound healing rates. Because of potential interactions of steroid hormones and immune function, we also measured plasma levels of testosterone and corticosterone. Mean healing rates differed significantly, with males in the darker high-elevation population healing more quickly compared to lighter low-elevation males. Males in the low-elevation population had significantly higher mean baseline steroid concentrations. These steroid hormones were also negatively associated with wound healing. We discuss potential differences in selective regime that could produce different patterns. These data also suggest that hormonal pleiotropy does not constrain phenotypic variation.

Phenotypic correlates of melanization in two Sceloporus occidentalis (Phrynosomatidae) populations: Behavior, androgens, stress reactivity, and ectoparasites.

Mechanisms underlying production of animal coloration can affect key traits besides coloration. Melanin, and molecules regulating melanin, can directly and indirectly affect other phenotypic traits including aggression, stress-reactivity, and immune function. We studied correlation of melanization with these other traits, comparing within- and between-population differences of adult male western fence lizards, Sceloporus occidentalis. We compared one high- and one low-elevation population in California where individuals are increasingly darker at higher elevations, working during comparable periods of the breeding season at each site (first egg clutch). We measured agonistic behaviors of free-ranging males in response to staged territorial intrusions (STIs). In other sets of males we measured baseline testosterone and corticosterone levels, and hormonal-reactivity to a stress handling paradigm. We counted ectoparasite loads for all males. There were no significant associations between individual variation in melanization and individual variation in any of the variables measured. However, analysis of behavior from the STIs revealed that males in the darker high-elevation population responded with more aggressive behavior compared to males in the lighter low-elevation population. Males in the low-elevation population had significantly higher mean baseline testosterone, but the two populations did not differ in adrenal function (baseline corticosterone or corticosterone after 1-h confinement stress). Males in the darker high-elevation population had higher mean mite loads compared to males in the lighter population. This array of phenotypic differences between the two populations, and the absence of trait associations when assessing individual variation, do not parallel the patterns in other vertebrates. We describe potential differences in selective regimes that could produce these different patterns across vertebrates. These data suggest that hormonal pleiotropy does not constrain phenotypic variation.

Influence of stressor duration on leukocyte and hormonal responses in male Southeastern five-lined skinks (Plestiodon inexpectatus).

Environmental stressors elicit elevations of plasma glucocorticoid hormones. Stressors also result in increases in the circulating heterophil to lymphocyte ratio (H:L ratio), which has been linked to increased glucocorticoids. Recently, there has been debate about the extent to which stress-induced elevations of corticosterone are tied to increases in the H:L ratio and about whether one measure of physiological stress might be used in lieu of the other. Few studies have examined the responses of both variables to standardized stressors in free-living animals, especially reptiles. Stressors may also affect levels of other hormones, such as prostaglandins or testosterone, that may influence leukocytes. Therefore, the H:L ratio, corticosterone, prostaglandin E(2) (PGE(2) ), and testosterone were monitored in free-living male southeastern five-lined skinks, Plestiodon inexpectatus, in response to various durations of confinement stress. Skinks confined for 1 hr had elevated H:L ratios and elevated corticosterone concentrations. In addition, corticosterone was positively correlated with the H:L ratio. In contrast, there were no effects of 2 hr of confinement on the H:L ratio, in spite of maximal corticosterone. Plasma PGE(2) was not affected by 15, 30, or 60 min of confinement but was positively correlated with monocytes and negatively correlated with lymphocytes in the 1 hr of confinement experiment. Consistent with many other studies of reptiles, confinement stress did not result in suppressed testosterone. Overall, the results indicate that, in reptiles at least, the H:L ratio and plasma corticosterone do not covary consistently and one measure of stress cannot be used in lieu of the other.