Arginine vasotocin-induced prostaglandin synthesis in vitro by the reproductive tract of the viviparous lizard Sceloporus jarrovi

Two experiments were performed to determine whether arginine vasotocin (AVT) stimulates synthesis of prostaglandins (PGs) in reptilian oviducts. Homogenized oviducal tissue from female Sceloporus jarrovi in early and late pregnancy were cultured with radiolabeled (14C) prostaglandin precursor, arachidonic acid (AA). In late pregnancy, oviducts exposed to AVT exhibited a greater conversion of AA to PGF2 alpha than did controls, whereas in early pregnancy there was no difference. The conversion of AA to other prostaglandins (PGA2, PGD2, PGE2, PGI2) was not influenced by AVT. The second experiment examined whether endogenous in vitro synthesis of PGF and PGE2 from intact, pregnant oviducts was stimulated by AVT (50 ng/ml; 100 ng/ml). Both doses of AVT induced a similar, significant rise in PGF concentrations within 30 min whereas no significant increase was noted in PGE2 concentrations until 90 min after treatment. Indomethacin pretreatment blocked synthesis of both PGF and PGE2 for 30 min following AVT treatment. These data indicate that AVT induces a highly specific rise in the synthesis of PGF from the oviduct of female S. jarrovi in late pregnancy. Furthermore, the prostaglandin-stimulating effect of AVT in reptiles appears homologous with the effect of oxytocin in mammals and AVT in birds. We hypothesize that this interaction is an evolutionarily conserved relationship found in all amniote vertebrates.     

Neurofilament and glycogen changes during cold acclimation in the trochlear nucleus of lizards (Sceloporus undulatus)

Brain Cell Biology - In lizards (Sceloporus undulatus), long term (13 or 19 weeks) acclimation to an environment of 6 °C produces a striking increase in the argyrophilic neurofibrillar network...     

An experimental manipulation of food and water: growth and time-space utilization of hatchling lizards (Sceloporus undulatus)

Summary At a site in western Nebraska, we studied the effects of supplemental food and water on growth, survival, and habitat utilization of hatchling iguanid lizards, Sceloporus undulatus. Hatchlings were randomly assigned to one of four experimental groups and received either (1) supplemental food only, (2) supplemental water only, (3) supplemental food and water, or (4) neither supplemental food nor water (control). Neither supplemental food nor supplemental water affected growth rate in length, growth rate in mass, or size-specific body mass (i.e. robustness) at the end of the treatment period. Over the 45 day treatment pierod, there were no significant differences among treatment groups in survival, home-range size, or habitat utilization. There were, however, significant differences in the index of activity. Hatchlings in the treatment groups receiving supplemental water were sighted more often than hatchlings in the control group. We conclude that, during our study, neither food nor water limited growth but that water limited activity.     

Changes in oviducal vascularity during the reproductive cycle of three oviparous lizards (Eumeces obsoletus, Sceloporus undulatus and Crotaphytus collaris)

Histologically derived estimates and ink suspension vascular casts were used to examine oviducal vascular changes. Vascularity peaked during gravidity and was correlated with maximal plasma progesterone concentrations. The vascular increase in the oviducal tissue was attributed exclusively to increased capillary densities. The greatest change occurred in the anterior uterus where incubation and egg shell secretion occur. Similar patterns of change in vascularity occurred in the infundibulum, although not as extreme as that seen in the anterior uterus, whereas no significant alterations were noted in the posterior uterus. These modifications mimic the pattern of vascular change occurring in viviparous lizards during simple placentation. We suggest that major changes in uterine vascularity may not be required for the evolution of simple chorioallantoic placentae in lizards.     

Testosterone inhibits growth in juvenile male eastern fence lizards (Sceloporus undulatus): implications for energy allocation and sexual size dimorphism

In the eastern fence lizard, Sceloporus undulatus, female-larger sexual size dimorphism develops because yearling females grow faster than males before first reproduction. This sexual growth divergence coincides with maturational increases in male aggression, movement, and ventral coloration, all of which are influenced by the sex steroid testosterone (T). These observations suggest that male growth may be constrained by energetic costs of activity and implicate T as a physiological regulator of this potential trade-off. To test this hypothesis, we used surgical castration and subsequent administration of exogenous T to alter the physiological and behavioral phenotypes of field-active males during the period of sexual growth divergence. As predicted, T inhibited male growth, while castration promoted long-term growth. Males treated with T also exhibited increased daily activity period, movement, and home range area. Food consumption did not differ among male treatments or sexes, suggesting that the inhibitory effects of T on growth are mediated by patterns of energy allocation rather than acquisition. On the basis of estimates derived from published data, we conclude that the energetic cost of increased daily activity period following T manipulation is sufficient to explain most (79%) of the associated reduction in growth. Further, growth may have been constrained by additional energetic costs of increased ectoparasite load following T manipulation. Similar studies of the proximate behavioral, ecological, and physiological mechanisms involved in growth regulation should greatly improve our understanding of sexual size dimorphism.     

Reproductive ecology of red-chinned lizards (Sceloporus undulatus erythrocheilus) in Southcentral Colorado: comparisons with other populations of a wide-ranging species

Summary We studied the reproductive ecology of a population of Sceloporus undulatus erythrocheilus near Walsenburg, CO during the summers of 1987–1990. Reproductive activity commences soon after emergence in early May and continues until the middle of July. Females mature in their second year following hatching at an age of 20–21 months and a size of 60–63 mm snout-vent length (SVL). Two clutches, averaging10.9 eggs, are produced per year. Larger females produce larger clutches with an increase of one egg per 3 mm SVL. Average SVL of females was 71 mm. Eggs are reasonably large (0.32 g) and relative clutch mass was the largest reported for the species (34% of body mass). A principal component analysis was used to examine relationships among reproductive characters in 12 populations of Sceloporus undulatus. The analysis suggested that patterns of reproduction in this wide-ranging species result from several factors reflecting both adaptive and phylogenetic sources of variation.     

Salmonellosis in laboratory-housed iguanid lizards (Sceloporus spp.).

Fifteen wild-caught iguanid lizards (14 Sceloporus variabilis and one S. malachiticus) were used in a 3 mo study on thermal acclimation. Over a 2 mo period, five of the lizards showed decreased activity, anorexia and enlarged joints, and were either found moribund or were euthanatized due to their poor condition. Specimens taken from lesions in four of the five lizards were cultured and were infected with Salmonella spp. Salmonella spp. was cultured from cloacal swabs in six of the 10 surviving lizards. Standard metabolic rates of those that were infected did not differ significantly from those that were not infected. We postulate that the lizards were inapparent carriers of Salmonella spp. at the time of capture and, as a result of stress, five developed active overwhelming systemic infections.     

Seasonal specificity of hormonal, behavioral, and coloration responses to within- and between-sex encounters in male lizards (Sceloporus undulatus).

This study reports the gender and seasonal specificity of hormonal, behavioral, and coloration responses displayed by "resident" male lizards (Sceloporus undulatus) exposed to male or female "intruders" during staged encounters in outdoor enclosures. Resident males were engaged in staged encounters with males or females for 1 h per day on 9 consecutive days during the breeding and postbreeding seasons. Male-specific responses occurred during the breeding but not the postbreeding season. These included (1) a transient increase in plasma testosterone (T) that was evident on Day 4 and had subsided by Day 10, (2) behavioral displays of aggression (full shows and chases), and (3) a lightening of dorsal integumental color. Female-specific behavioral responses (nod sets) were displayed in both seasons. Season-specific responses consisted only of a transient increase in plasma corticosterone (B) during the breeding season that was evident on Day 4 and had subsided by Day 10. Pushups were displayed in response to both genders during both seasons, although the frequency of pushups was significantly higher in response to females than to males during the postbreeding season. The coloration of residents did not change in response to male intruders during the postbreeding season or to females during either season. These results define the gender and seasonal specificity of hormonal, behavioral, and coloration responses of resident male S. undulatus in social interactions with conspecifics. Thus, our results clarify the biological significance of these responses in terms of potentially aggressive versus courtship interactions and breeding versus postbreeding contexts.     

The Ontogeny of Display Behaviour in Sceloporus undulatus: (Sauria: Iguanidae)

A longitudinal study (from neonate to adult) was made of the display behaviour of 36 fence lizards from 8 clutches. A frame-by-frame analysis of 941 video-taped displays revealed that the hatchlings possessed almost all of their adult display patterns by the first two days of life. Of the total variance in the display behaviour, 7% or less was attributed to experience. Of the variance among lizards, approximately half came from between clutches and half from between clutchmates. Our data indicate that the visual communication signals (head bob displays) of this vertebrate species are almost totally innate.     

Characterization of gastrointestinal chitinase in the lizard Sceloporus undulatus garmani (Reptilia: Phrynosomatidae)

Most studies on chitinase activity in lizards have been concerned with Palaearctic (European) and Laurasian (Middle Eastern and Asian) taxa. Several genera of Old World lizards, Anguis, Uromastix, Chamaeleo and Lacerta, have been shown to possess chitinolytic activity. To date, only one New World lizard, Anolis carolinensis, has been reported to exhibit chitinolytic activity. In the present study, chitinase activity was characterized in a second New World taxon, Sceloporus undulatus garmani, a New World, phrynosomatid lizard. Chitinolytic activity was measured by incubating tissue extracts with a radioactive chitin substrate, acetyl-[H³]chitin and determining acid soluble radioactivity as an estimate for chitin hydrolysis. Chitinolytic activity was present in stomach, small intestine and pancreas extracts, with the stomach and pancreas having the highest specific activities. Chitinolytic activity was higher at pH 4.5 than at pH 7.5. The stomach chitinase is immunologically similar to the gastric chitinase previously described for rainbow trout. Western blot analysis showed anti-chitinase cross-reactivity in the extracts of the stomach, but no cross-reactivity in the pancreatic or intestinal extracts, suggesting different isoforms of chitinase. There was no detected lysozyme activity (less than 0.01 mg/ml lysozyme) present in the extracts of the stomach, small intestine and pancreas. The localization of chitinolytic activity in S. u. garmani is in agreement with earlier reptilian reports on the distribution of chitinase.     

Scanning electron microscopy of the placental interface in the viviparous lizard Sceloporus jarrovi (Squamata: Phrynosomatidae)

Placental membranes mediate maternal‐fetal exchange in all viviparous reptilian sauropsids. We used scanning electron microscopy to examine the placental interface in the mountain spiny lizard, Sceloporus jarrovi (Phrynosomatidae). From the late limb bud stage until birth, the conceptus is surrounded by placental membranes formed from the chorioallantois and yolk sac omphalopleure. The chorioallantois lies directly apposed to the uterine lining with no intervening shell membrane. Both fetal and maternal sides of the chorioallantoic placenta are lined by continuous layers of flattened epithelial cells that overlie dense capillary networks. The chorioallantoic placenta shows specializations that enhance respiratory exchange, as well as ultrastructural evidence of maternal secretion and fetal absorption. The yolk sac placenta contains enlarged fetal and maternal epithelia with specializations for histotrophic nutrient transfer. This placenta lacks intrinsic vascularity, although the vascular allantois lies against its inner face, contributing to an omphallantoic placenta. In a specialized region at the abembryonic pole, uterine and fetal tissues are separated by a compact mass of shed shell membrane, yolk droplets, and cellular debris. The omphalopleure in this region develops elongate folds that may contribute to sequestration and absorption of this material. Fetal membrane morphogenesis and composition in S. jarrovi are consistent with those of typical squamates. However, this species exhibits unusual placental specializations characteristic of highly placentotrophic lizards. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.     

Convergent evolution of embryonic growth and development in the eastern fence lizard (Sceloporus undulatus)

Theory predicts that cold environments will select for strategies that enhance the growth of ectotherms, such as early emergence from nests and more efficient use of resources. We used a common garden experiment to detect parallel clines in rates of embryonic growth and development by eastern fence lizards (Sceloporus undulatus). Using realistic thermal conditions, we measured growth efficiencies and incubation periods of lizards from five populations representing two distinct clades. In both clades, embryos from cold environments (Indiana, New Jersey, and Virginia) grew more efficiently and hatched earlier than embryos from warm environments (Florida and South Carolina). Because eggs from cold environments were larger than eggs from warm environments, we experimentally miniaturized eggs from one population (Virginia) to determine whether rapid growth and development were caused by a greater maternal investment. Embryos in miniaturized eggs grew as efficiently and incubated for the same duration as embryos in unmanipulated eggs. Taken together, our results suggest countergradient variation has evolved at least twice in S. undulatus.     

Physaloptera retusa (Nematoda, Physalopteridae) in naturally infected sagebrush lizards, Sceloporus graciosus (Iguanidae)

Of 292 adult sagebrush lizards (Sceloporus graciosus) examined for the stomach nematode, Physaloptera retusa, 81 (28%) were infected. Attached nematodes caused erosive inflammatory foci in the stomach mucosa. Early lesions were U-shaped. Degenerating gastric glands occurred in the adjacent mucosa. Healed lesions were fibrous connective tissue scars; regenerating gastric glands occurred at the periphery. There were no externally visible signs of infection.     

Delimiting species using DNA and morphological variation and discordant species limits in spiny lizards (Sceloporus)

Haplotype phylogenies based on DNA sequence data are increasingly being used to test traditional species-level taxonomies based on morphology. However, few studies have critically compared species limits based on morphological and DNA data, and the methods used to delimit species using either type of data are only rarely explained. In this paper, we review three approaches for species delimitation (tree-based with DNA data and tree-based and character-based with morphological data) and propose explicit protocols for each. We then compare species limits inferred from these approaches, using morphological and mtDNA data for the Yarrow's spiny lizard (Sceloporus jarrovii), a traditionally polytypic species from the southwestern United States and Mexico. All three approaches support division of S. jarrovii into five species, but only two species are the same among the three approaches. We find the greatest support for the five species that are delimited based on mtDNA data, and we argue that mtDNA data may have important (and previously unappreciated) advantages for species delimitation. Because different data and approaches can disagree so extensively, our results demonstrate that the methodology of species delimitation is a critical issue in systematics.     

Hormones, performance and fitness: Natural history and endocrine experiments on a lizard (Sceloporus undulatus)

We used the "morphology-performance-fitness" paradigm (Arnold, 1983) as our framework to investigate endocrine control of performance and fitness in Sceloporus undulatus (Eastern Fence Lizard). Focusing on males, we used the "natural experiments" of seasonal, sexual, and developmental variation in growth and in exercise endurance to identify testosterone and corticosterone as potential modulators of performance and related traits of interest. We followed with experimental manipulations of testosterone to investigate functional relationships, both in the laboratory and in the field. Further, we used focal observations and demographic studies, coupled with genetic determination of paternity, to test associations between performance and fitness, measured as reproductive success. We found that in males, endurance and plasma concentrations of testosterone and corticosterone are at their peaks in the spring breeding season, when lizards are most actively engaged in patrolling home ranges and in reproductive behavior. At that time, plasma concentrations of testosterone are correlated with body size; plasma concentrations of corticosterone and parameters of home range, including area and the number of overlapped females, are correlated with home-range overlap between males and females. During prereproductive development, males (but not females) experience a maturational increase in plasma testosterone. At about the same time, they become more active, expand their home ranges, and grow less quickly than do females, suggesting a trade-off in the allocation of energy, mediated by testosterone. Experimentally, testosterone has positive effects on fitness by stimulating endurance and reproductive activity and increasing home-range area, but it exacts costs in fitness by increasing ectoparasitism, decreasing growth, and decreasing survivorship. We found evidence of selection on body size, endurance, and home-range size (and thus access to potential mates). Despite having positive effects on performance traits, plasma concentrations of testosterone were not correlated with number of offspring sired by males. However, we found a strong correlation between the level of plasma corticosterone and the number of offspring sired. We also found evidence of size-assortative mating, indicating that for males, both the number and the size (and thus, fecundity) of their mates increase with body size. Our studies exemplify the power of natural history combined with experimental endocrine manipulations to identify hormonal regulators of performance and linkages to fitness. Furthermore, our results illustrate ecological and evolutionary significance of individual variation in endocrine traits.     

Genetic, prenatal, and postnatal correlates of dispersal in hatchling fence lizards (Sceloporus occidentalis)

Similarity of dispersal behavior among siblings is common invertebrates. However, little is known about the factors (genetic,prenatal, postnatal) generating this similarity. Here we analyzedpotential influences on the dispersal patterns of multiple familiesof hatchling fence lizards, Sceloporus occidentalis. We capturednear-term females from the field, incubated their eggs in thelaboratory, measured various traits of the hatchlings and dams,and then released the hatchlings at a number of sites in nature.We recaptured hatchlings 5–6 weeks later and measuredthe direct distance to the release site. Because we treatedhatchlings (from eggs to release) randomly with respect to sibship,we eliminated the possibility that any observed sibling similarityin dispersal is merely an artifact of common postnatal influences.To analyze dispersal, we developed a new method that does notmake an arbitrary choice of a threshold distance separatingdispersers from nondispersers. We found a significant familyeffect on dispersal. We suspect that this family effect originatesfrom genetic influences rather than from prenatal ones. Indeed,hatchling dispersal was remarkably unrelated to numerous traits(of clutches, mothers, or hatchlings) that might reflect prenataleffects. However, we did find that males were more likely todisperse than females, as predicted for polygynous species.Finally, characteristics of the release site did not appearto influence dispersal.     

Pterin-based ornamental coloration predicts yolk antioxidant levels in female striped plateau lizards (Sceloporus virgatus)

1. Maternal investment in egg quality can have important consequences for offspring fitness. For example, yolk antioxidants can affect embryonic development as well as juvenile and adult phenotype. Thus, females may be selected to advertise their yolk antioxidant deposition to discriminatory males via ornamental signals, perhaps depending on the reproductive costs associated with signal production. 2. Female striped plateau lizards (Sceloporus virgatus) develop pterin-based orange colour patches during the reproductive season that influence male behaviour and that are positively associated with the phenotypic quality of the female and her offspring. Here, we assessed one potential developmental mechanism underlying the relationship between offspring quality and female ornamentation in S. virgatus, by examining the relationship between ornament expression and yolk antioxidant levels. 3. As expected, concentrations of the yolk antioxidants vitamin A, vitamin E and carotenoids (lutein and zeaxanthin) were strongly positively intercorrelated. Eggs from larger clutches had fewer antioxidants than eggs from smaller clutches, suggesting that females may be limited in antioxidant availability or use. Fertilized and unfertilized eggs did not differ in yolk antioxidant levels. 4. The size of a female's ornament was positively related to both the concentration and total amount of yolk antioxidants, and ornament colour was positively related to yolk antioxidant concentration. Thus, in S. virgatus, female ornaments may advertise egg quality. In addition, these data suggest that more ornamented females may produce higher-quality offspring, in part because their eggs contain more antioxidants. As the colour ornament of interest is derived from pterins, not carotenoids, direct resource trade-offs between ornaments and eggs may be eliminated, reducing reproductive costs associated with signalling. 5. This is the first example of a positive relationship between female ornamentation and yolk antioxidants in reptiles and may indicate the general importance of these patterns in oviparous vertebrates.     

Locomotor performance of hatchling fence lizards (Sceloporus occidentalis): Quantitative genetics and morphometric correlates

Summary We examined heritabilities and correlations among measures of locomotor performance (speed, stamina) and among possible morphometric determinants of performance (hindlimb span, tail length) in families of hatchling lizards (Sceloporus occidentalis). We were particularly interested in determining whether these traits were heritable and thus might potentially respond genetically to selection. Moreover, we wished to determine whether speed and stamina are negatively genetically correlated, as suggested bya priori physiological and empirical considerations. All four traits appeared to be significantly heritable. Broadsense heritabilities were 0.33–0.36 for speed, 0.35–0.36 for stamina, 0.45–0.51 for hindlimb span, and 0.46–0.47 for tail length. Contrary to expectations, speed and stamina were not negatively genetically correlated. Hindlimb span and tail length, however, were negatively genetically correlated (but not phenotypically correlated). Hindlimb span and stamina were positively phenotypically correlated. Thus, for example, selection for longer hindlimb span could potentially result in shorter tails, contrary to evolutionary predictions based only on phenotypic correlations.     

Seasonal alterations in adrenocortical cell function associated with stress-responsiveness and sex in the eastern fence lizard (Sceloporus undulatus)

We characterized steroidogenic properties of dispersed adrenocortical cells from field-active male and female eastern fence lizards (Sceloporus undulatus) to investigate whether alterations in cell function could, in part, explain seasonal variation in baseline and stress-induced plasma corticosterone (B). Lizards were collected during the breeding and postbreeding seasons and shortly prior to hibernation. Dispersed cells in vitro produced B, aldosterone (ALDO), and progesterone in response to 8-Br-cAMP, 25-(OH)cholesterol, adrenocorticotropin (ACTH; as little as 100 fM), and angiotensin II. Maximal progesterone, B, and ALDO responses to ACTH were roughly 1000%, 500%, and 100% greater than corresponding basal values. Angiotensin II was an effective steroidogenic stimulant but much less so than ACTH. Corticosteroid production exhibited considerable steroid-specific variation among seasons. Maximal ACTH-induced B production was lower in the postbreeding season than at either of the other two measurement points, essentially opposite to the pattern for ALDO. Males and females generally produced B at similar rates, but ALDO and progesterone showed numerous sex differences that usually covaried between the two steroids. Cellular sensitivity to 25-(OH)cholesterol and angiotensin II showed few sex differences or seasonal changes. In contrast, sensitivity to ACTH decreased markedly from the breeding to the postbreeding season in males, corresponding to the decrease in stress-responsiveness, and in both sexes was considerably lower prior to hibernation than during the breeding season. Under some conditions, plasma B may be limited by the production capacity of adrenocortical cells. In summary, seasonal variations in body condition, reproductive activity, and baseline and stress-induced plasma B may be attributed at least in part to alterations in adrenocortical cell steroidogenic function.