Lukáš Kubička

Cell Size but Not Genome Size Affects Scaling of Metabolic Rate in Eyelid Geckos

The metabolic theory of ecology (MTE) predicts the ubiquity of the of 3/4 scaling exponent relating metabolic rate (MR) to body mass, as well as cell‐size invariance coupled with body‐size dependence of cellular MR in quickly dividing cells. An alternative prediction is that MR scales interspecifically with a coefficient that is between 2/3 and 1, depending on the cell size and cell MR, which is mostly driven by the cell surface‐to‐volume ratio. We tested (1) the contribution of cell size to interspecific differences in MR and (2) whether the cell size–MR relationship is mediated by genome size (GS), which usually correlates positively with cell size. We tested (1) and (2) using erythrocyte area as a proxy for cell size in 14 eyelid geckos, which belong to a monophyletic group exhibiting large body‐size variation. The scaling of standard MR (SMR) was significantly lower than 3/4, whereas mass‐specific SMR correlated with erythrocyte area in both phylogenetically adjusted and conventional analyses. This points to cell‐size variation as the factor governing metabolic rate scaling, which questions predictions of the MTE. However, the nonsignificance of the correlation between mass‐specific SMR and GS undermines the strength of the relation between GS and cell size, at least in these species.

Macroevolutionary pattern of sexual size dimorphism in geckos corresponds to intraspecific temperature-induced variation

Many animal lineages exhibit allometry in sexual size dimorphism (SSD), known as ‘Rensch’s rule’. When applied to the interspecific level, this rule states that males are more evolutionary plastic in body size than females and that male‐biased SSD increases with body size. One of the explanations for the occurrence of Rensch’s rule is the differential‐plasticity hypothesis assuming that higher evolutionary plasticity in males is a consequence of larger sensitivity of male growth to environmental cues. We have confirmed the pattern consistent with Rensch’s rule among species of the gecko genus Paroedura and followed the ontogeny of SSD at three constant temperatures in a male‐larger species (Paroedura picta). In this species, males exhibited larger temperature‐induced phenotypic plasticity in final body size than females, and body size and SSD correlated across temperatures. This result supports the differential‐plasticity hypothesis and points to the role phenotypic plasticity plays in generating of evolutionary novelties.

Does the mechanism of sex determination constrain the potential for sex manipulation? A test in geckos with contrasting sex-determining systems

The concentration of yolk steroids was suggested to influence offspring gender in oviparous animals subject to both temperature-dependent sex determination (TSD) and genotypic sex determination (GSD). However, the proposed mechanisms of steroid effects are thought to differ between TSD and GSD: a direct effect of oestrogens on gonad feminisation in TSD species vs a differential induction of male-producing or female-producing gametes in GSD species. Geckos offer an ideal opportunity for testing these suggested mechanisms. Closely related gecko species differ in their modes of sex determination. They lay clutches of two synchronously formed eggs; both eggs share equal steroid levels. If identical hormonal composition and environment during vitellogenesis, gravidity and incubation determine the sex of the progeny, siblings should share the same gender in both TSD and GSD geckos. We found strong support for this prediction in a TSD gecko species. Among clutches that were incubated at the temperature that produced both sexes, there were no clutches with siblings of the opposite sex. On the other hand, about half of the clutches yielded siblings of the opposite sex in four GSD species. These results suggest that sex-determining systems constrain the ability of the female to produce single-sex siblings and, hence, it seems that the GSD mechanism constrains the opportunities for sex ratio manipulation in geckos via yolk steroid manipulation.

Neither male gonadal androgens nor female reproductive costs drive development of sexual size dimorphism in lizards

SUMMARY Sexual size dimorphism (SSD) is an extensively studied phenomenon in animals, including reptiles, but the proximate mechanism of its development is poorly understood. The most pervasive candidates are: (1) androgen-mediated control of growth, i.e. a positive effect of gonadal androgens (testosterone) on male growth in male-larger species, and a negative effect in female-larger species; and (2) sex-specific differences in energy allocation to growth, e.g. sex with larger reproductive costs should result in smaller body size. We tested these hypotheses in adults of the male-larger lizard Paroedura picta by conducting castrations with and without testosterone implants in males and manipulating reproductive status in females. Castration or testosterone replacement had no significant effect on final body length in males. High investment to reproduction had no significant effect on final body length in intact females. Interestingly, ovariectomized females and females with testosterone implants grew to larger body size than intact females. We did not find support for either of the above hypotheses and suggest that previously reported effects of gonadal androgens on growth in male lizards could be a consequence of altered behaviour or social status in manipulated individuals. Exogenous testosterone in females led to decreased size of ovaries; its effect on body size may be caused by interference with normal ovarian function. We suggest that ovarian factors, perhaps estrogens, not reproductive costs, can modify growth in female lizards and may thus contribute to the development of SSD. This hypothesis is largely supported by published results on the effect of testosterone treatment or ovariectomy on body size in female squamates.

Elevated testosterone is required for male copulatory behavior and aggression in Madagascar ground gecko (Paroedura picta)

Elevated levels of gonadal androgens are often required for the expression of male-specific behavioral and morphological traits in all classes of vertebrates, including reptiles. Here, we tested the role of male gonadal androgens in the control of male sexual behavior, aggressive behavior, and size of the hemipenes in the gecko Paroedura picta. We performed hormonal manipulations involving castration with and without testosterone (T) replacement in males and application of exogenous T and ovariectomy in females. Castration suppressed sexual behavior and hemipenes size in males, and these effects were fully rescued by exogenous T. Sexual behavior and growth of the hemipenes were masculinized by male-typical levels of T in females, while ovariectomized females retained female-typical expression of these traits. These results indicate that the development of male sexual behavior in adult females does not require early or pubertal organization. Elevated T increased the likelihood of aggressive behavior directed toward a male intruder, but aggression occurred only rarely. Elevated T is necessary and sufficient for enlargement of the hemipenes and the expression of male sexual behavior in both males and females of Paroedura picta. In contrast to sexual behavior, the expression of aggressive behavior is apparently more dependent on other factors in addition to T itself.

Ontogeny of pronounced female-biased sexual size dimorphism in the Malaysian cat gecko (Aeluroscalabotes felinus: Squamata: Eublepharidae): A test of the role of testosterone in growth regulation

Species differences in the effect of male gonadal androgens on male growth are considered a possible mechanism allowing shifts in magnitude and even direction of sexual size dimorphism (SSD) in squamate reptiles. According to the bipotential growth regulation hypothesis, the androgen testosterone (T) enhances male growth in species with male-biased SSD and conversely inhibits male growth in males of female-larger species. In the present study, we describe the ontogeny of the pronounced female-biased SSD and report the effect of T on growth via hormonal manipulations in males and females of the Malaysian cat gecko (Aeluroscalabotes felinus). In accord with the predictions of the bipotential growth regulation hypothesis, growth was inhibited by replacement of T in castrated males. Additionally, exogenous T inhibited growth of females to male-typical levels. Nevertheless, male castration alone did not significantly affect growth, contrary to the prediction of the bipotential growth regulation hypothesis, which contradicts the generality of this hypothesis. Application of exogenous T to females can interfere with normal ovarian function. Therefore, although not directly tested in this study, we suggest that ovarian effects on the ontogeny of SSD in A. felinus are consistent with our results. The development of SSD is a function of differential growth between the sexes, and potential sex-specific growth regulation in both males and females should be taken into account as possible proximate mechanisms responsible for SSD.

Endogenous control of sexual size dimorphism: Gonadal androgens have neither direct nor indirect effect on male growth in a Madagascar ground gecko (Paroedura picta).

Changes in the effect of gonadal androgens on male growth are considered as a possible mechanism allowing shifts in magnitude and even direction of sexual size dimorphism in vertebrates, particularly squamate reptiles. Positive effects of gonadal androgens on male growth were found in several male-larger species of lizards. Contrastingly, we document that in the male-larger Madagascar ground gecko (Paroedura picta) gonadal androgens do not affect male growth under constant thermal conditions. However, the absence of a thermal gradient might prevent the potential indirect effect of gonadal androgens on growth via the influence of circulating hormones on an individuals thermoregulation and hence metabolic rate. In order to study this, we monitored the growth and body temperature of socially isolated sham-operated and castrated males of the same species in a thermal gradient. We also compared the oxygen consumption and activity between the treatment groups in the open field to test the effect of gonadal hormones on these traits potentially affecting growth. Even under a thermal gradient we found no effect of gonadal androgens on growth rate or final body dimensions. Castration also did not significantly affect oxygen consumption or activity in the open field test. Together with our previous findings, we can exclude both the direct effect of male gonadal androgens on the ontogeny of sexual size dimorphism via the influence on the growth axis, and the indirect influence of gonadal androgens acting on the ontogeny of SSD through the effect on thermoregulation, metabolic rate and activity.

Ovarian control of growth and sexual size dimorphism in a male-larger gecko

ABSTRACT Sexual size dimorphism (SSD) reflects sex-specific solutions to the allocation of energy among growth, reproduction and survival; however, the proximate mechanisms behind these solutions are still poorly known even in vertebrates. In squamates, sexual differences in body size used to be attributed to direct energy allocation to energetically demanding processes, largely to reproduction. In addition, SSD is assumed to be controlled by specific endogenous mechanisms regulating growth in a sex-specific manner, namely masculinization by male gonadal androgens or feminization by ovarian hormones. We performed a manipulative growth experiment in females of the male-larger gecko Paroedura picta in order to test the reproductive cost hypothesis, the male androgen hypothesis and the ovarian hormone hypothesis. Specifically, we investigated the effect of total ovariectomy, prepubertal ovariectomy, unilateral ovariectomy, and total ovariectomy followed by exogenous estradiol, dihydrotestosterone or testosterone treatment, on female growth in comparison to males and reproductively active females. The present results and the results of our previous experiments do not support the hypotheses that SSD reflects direct energy allocation to reproduction and that male gonadal androgens are involved. However, all lines of evidence, particularly the comparable growth of reproducing intact and unilaterally ovariectomized females, were concordant with the control of SSD by ovarian hormones. We suggest that feminization of growth by female gonadal hormones should be taken into consideration as an endogenous pathway responsible for the ontogeny of SSD in squamates. Summary: Sexual size dimorphism in a male-larger lizard is influenced by neither male gonadal androgens nor female reproductive allocation, but instead is controlled by ovarian hormones.

Androgenic control of male-typical behavior, morphology and sex recognition is independent of the mode of sex determination: A case study on Lichtenfelder's gecko (Eublepharidae: Goniurosaurus lichtenfelderi).

Previous work on lizards has shown that many sexually dimorphic traits depend on testosterone (T), but the details of this control can vary among species. Here, we tested the role of T on the expression of morphological, physiological, and behavioral traits in Lichtenfelders gecko (Goniurosaurus lichtenfelderi), from the lizard family Eublepharidae notable for interspecific variation in sexually dimorphic traits and the mode of sex determination. Experiments included three groups of males (intact control, surgically castrated, castrated with T replacement) and two groups of females (intact control, T supplemented). In males, castration caused reductions in 1) the size of hemipenes, 2) offensive aggression, 3) male sexual behavior in a neutral arena, 4) activity of precloacal glands, and 5) loss of male chemical cues for sex recognition. These reductions were not observed in castrated males with T replacement. Interestingly, castrated males performed sexual behavior in their home cages, which shows that the effect of T depends on the environmental context. Notably, tail vibration, previously reported as a courtship behavior in other eublepharids, is displayed by males of G. lichtenfelderi during interactions with conspecifics of both sexes, suggesting an evolutionary shift in the meaning of this signal. In females, T induced growth of hemipenes and male-typical courtship but did not induce precloacal pore activity, aggression, or mounting. In comparison to previous reports on Eublepharis macularius, our results indicate that effects of T do not depend on the mode of sex determination. Further, our results extend our understanding of the complexity of control of male traits and illustrate how lability in the effects of T can be a general mechanism causing evolutionary changes in the components of suites of functionally correlated traits.

Overlap of Female Reproductive Cycles Explains Shortened Interclutch Interval in a Lizard with Invariant Clutch Size (Squamata: Gekkonidae: Paroedura picta)

Variable clutch size is unambiguously an ancestral state in reptiles. Only several lizard lineages have evolved so-called invariant clutch size, where all females lay just one or two eggs per clutch. This mode of reproduction is characteristic for geckos. In some gecko lineages, decreased fecundity in a single clutch is compensated by conspicuous shortening of interclutch intervals. The proximate mechanism of high clutch frequency in these geckos is not known. Here, we document that three subsequently laid clutches develop simultaneously in females of the Madagascar ground gecko (Paroedura picta). The extremely short interclutch intervals in this species—even as short as a week—thus could be attributed to the overlap of female reproductive cycles. Such overlap should be associated with altered female hormonal cycles. Based on measurements of hormonal levels, we suggest that cycles of estradiol and progesterone during reproductive cycles of females in P. picta are largely independent. Thus, in contrast to the presumable ancestral reptile state, higher levels of progesterone do not seem to interfere with vitellogenesis in this species. We discuss potential consequences of this derived mode of reproduction, such as possible simultaneous maternal transfer of nutrients and other yolk components to several subsequent clutches.