Matthew B. Lovern

FASTER LIZARDS SIRE MORE OFFSPRING: SEXUAL SELECTION ON WHOLE-ANIMAL PERFORMANCE

Sexual selection operates by acting on variation in mating success. However, since selection acts on wholeorganism manifestations (i.e., performance) of underlying morphological traits, tests for phenotypic effects of sexual selection should consider whole-animal performance as a substrate for sexual selection. Previous studies have revealed positive relationships between performance and survival, that is, natural selection, but none have explicitly tested whether performance may influence reproductive success (through more matings), that is, sexual selection. Performance predicts dominance in some species, implying the effects of sexual selection, but how it does so has not been established, nor is it certain whether performance might be a by-product of selection for something else, for example, elevated circulating testosterone levels. We investigated the potential for sexual selection on sprint speed performance in collared lizards (Crotaphytus collaris), considering the potential mediating effects of circulating hormone levels. Among territorial, adult male collared lizards, only sprint speed significantly predicted territory area and number of offspring sired as determined by genetic paternity analysis. Body size, head size, and hind limb length had no effect. Neither plasma testosterone levels nor corticosterone levels correlated with sprint speed, territory area, or number of offspring sired. Thus, our results provide a direct link between whole-animal performance and reproductive success, suggesting that intrasexual selection can act directly on sprint speed performance and drive the evolution of underlying morphological traits.

Maternal nutrition affects reproductive output and sex allocation in a lizard with environmental sex determination

Life-history traits such as offspring size, number and sex ratio are affected by maternal feeding rates in many kinds of animals, but the consequences of variation in maternal diet quality (rather than quantity) are poorly understood. We manipulated dietary quality of reproducing female lizards (Amphibolurus muricatus; Agamidae), a species with temperature-dependent sex determination, to examine strategies of reproductive allocation. Females maintained on a poor-quality diet produced fewer clutches but massively (twofold) larger eggs with lower concentrations of yolk testosterone than did conspecific females given a high-quality diet. Although all eggs were incubated at the same temperature, and yolk steroid hormone levels were not correlated with offspring sex, the nutrient-deprived females produced highly male-biased sex ratios among their offspring. These responses to maternal nutrition generate a link between sex and offspring size, in a direction likely to enhance maternal fitness if large body size enhances reproductive success more in sons than in daughters (as seems plausible, given the mating system of this species). Overall, our results show that sex determination in these animals is more complex, and responsive to a wider range of environmental cues, than that suggested by the classification of ‘environmental sex determination’.

Elevated plasma corticosterone increases metabolic rate in a terrestrial salamander

Plasma glucocorticoid hormones (GCs) increase intermediary metabolism, which may be reflected in whole-animal metabolic rate. Studies in fish, birds, and reptiles have shown that GCs may alter whole-animal energy expenditure, but results are conflicting and often involve GC levels that are not physiologically relevant. A previous study in red-legged salamanders found that male courtship pheromone increased plasma corticosterone (CORT; the primary GC in amphibians) concentrations in males, which could elevate metabolic processes to sustain courtship behaviors. To understand the possible metabolic effect of elevated plasma CORT, we measured the effects of male courtship pheromone and exogenous application of CORT on oxygen consumption in male red-legged salamanders (Plethodon shermani). Exogenous application of CORT elevated plasma CORT to physiologically relevant levels. Compared to treatment with male courtship pheromone and vehicle, treatment with CORT increased oxygen consumption rates for several hours after treatment, resulting in 12% more oxygen consumed (equivalent to 0.33 J) during our first 2h sampling period. Contrary to our previous work, treatment with pheromone did not increase plasma CORT, perhaps because subjects used in this study were not in breeding condition. Pheromone application did not affect respiration rates. Our study is one of the few to evaluate the influence of physiologically relevant elevations in CORT on whole-animal metabolism in vertebrates, and the first to show that elevated plasma CORT increases metabolism in an amphibian.

The maternal environment affects offspring viability via an indirect effect of yolk investment on offspring size.

Environmental conditions that reproductive females experience can influence patterns of offspring provisioning and fitness. In particular, prey availability can influence maternal reproduction and, in turn, affect the viability of their offspring. Although such maternal effects are widespread, the mechanisms by which these effects operate are poorly understood. We manipulated the amount of prey available to female brown anole lizards (Anolis sagrei) to evaluate how this factor affects patterns of reproductive investment (total egg output, egg size, yolk steroids) and offspring viability (morphology, growth, survival). Experimental reduction of yolk in a subset of eggs enabled us to evaluate a potential causal mechanism (yolk investment) that mediates the effect of maternal prey availability on offspring viability. We show that limited prey availability significantly reduced egg size, which negatively influenced offspring size, growth, and survival. Experimental yolk removal from eggs directly reduced offspring size, which, in turn, negatively affected offspring growth and survival. These findings show that maternal environments (i.e., low prey) can affect offspring fitness via an indirect effect of yolk investment on offspring size and highlight the complex set of indirect effects by which maternal effects can operate.

Evolutionary origin of Tbr2-expressing precursor cells and the subventricular zone in the developing cortex

The subventricular zone (SVZ) is greatly expanded in primates with gyrencephalic cortices and is thought to be absent from vertebrates with three‐layered, lissencephalic cortices, such as the turtle. Recent work in rodents has shown that Tbr2‐expressing neural precursor cells in the SVZ produce excitatory neurons for each cortical layer in the neocortex. Many excitatory neurons are generated through a two‐step process in which Pax6‐expressing radial glial cells divide in the VZ to produce Tbr2‐expressing intermediate progenitor cells, which divide in the SVZ to produce cortical neurons. We investigated the evolutionary origin of SVZ neural precursor cells in the prenatal cerebral cortex by testing for the presence and distribution of Tbr2‐expressing cells in the prenatal cortex of reptilian and avian species. We found that mitotic Tbr2+ cells are present in the prenatal cortex of lizard, turtle, chicken, and dove. Furthermore, Tbr2+ cells are organized into a distinct SVZ in the dorsal ventricular ridge (DVR) of turtle forebrain and in the cortices of chicken and dove. Our results are consistent with the concept that Tbr2+ neural precursor cells were present in the common ancestor of mammals and reptiles. Our data also suggest that the organizing principle guiding the assembly of Tbr2+ cells into an anatomically distinct SVZ, both developmentally and evolutionarily, may be shared across vertebrates. Finally, our results indicate that Tbr2 expression can be used to test for the presence of a distinct SVZ and to define the boundaries of the SVZ in developing cortices. J. Comp. Neurol. 524:433–447, 2016.

Social regulation of adult neurogenesis in a eusocial mammal

The present study examined the effects of social status on adult neurogenesis in an extreme cooperative breeder: the naked mole rat. These animals live in large colonies of up to 300 individuals, with a strict reproductive dominance hierarchy; one female and one to three males breed, and all other members are socially subordinate and reproductively suppressed. We examined the effects of social and gonadal cues on doublecortin (DCX; a marker for immature neurons) immunoreactivity in the dentate gyrus (DG), piriform cortex (PCx) and basolateral amygdala (BLA) by comparing dominant breeding animals to non-breeding subordinates from intact colonies. We also examined DCX expression in subordinate animals that had been removed from their colony and paired with an opposite- or same-sex conspecific for 6months. Compared to subordinates, dominant breeders had significantly reduced DCX immunoreactivity in all brain areas, with BLA effects confined to females. By contrast, the effects of same- versus opposite-sex housing were region-specific. In the DG and PCx, more DCX immunoreactivity was observed for opposite- than same-sex-paired subordinates. Conversely, same-sex-paired females had more DCX immunoreactivity than opposite-sex-paired females in the BLA. Gonadectomy did not affect DCX expression in opposite-sex-paired animals, and no significant relationships between gonadal steroids and DCX immunoreactivity were detected, suggesting that group differences in neurogenesis are independent of gonadal hormones. The apparent lower neurogenic capacity displayed by breeders contrasts previous reports on neurogenesis and social rank, challenging the conventional view that subordination is stressful and impairs neurogenesis. Future work will clarify whether the present findings can be attributed to status-dependent differences in stress, behavioral plasticity, or life stage.

Socially regulated reproductive development: analysis of GnRH-1 and kisspeptin neuronal systems in cooperatively breeding naked mole-rats (Heterocephalus glaber).

In naked mole‐rat (NMR) colonies, breeding is monopolized by the queen and her consorts. Subordinates experience gonadal development if separated from the queen. To elucidate the neuroendocrine factors underlying reproductive suppression/development in NMRs, we quantified plasma gonadal steroids and GnRH‐1‐ and kisspeptin‐immunoreactive (ir) neurons in subordinate adults and in those allowed to develop into breeders, with or without subsequent gonadectomy. In males and females, respectively, plasma testosterone and progesterone are higher in breeders than in subordinates. No such distinction occurs for plasma estradiol; its presence after gonadectomy and its positive correlation with adrenal estradiol suggest an adrenal source. Numbers of GnRH‐1‐ir cell bodies do not differ between gonad‐intact breeders and subordinates within or between the sexes. As in phylogenetically related guinea pigs, kisspeptin‐ir processes pervade the internal and external zones of the median eminence. Their distribution is consistent with actions on GnRH‐1 neurons at perikaryal and/or terminal levels. In previously investigated species, numbers of kisspeptin‐ir cell bodies vary from substantial to negligible according to sex and/or reproductive state. NMRs are exceptional: irrespective of sex, reproductive state, or presence of gonads, substantial numbers of kisspeptin‐ir cell bodies are detected in the rostral periventricular region of the third ventricle (RP3V) and in the anterior periventricular (PVa), arcuate, and dorsomedial hypothalamic nuclei. Nevertheless, the greater number in the RP3V/PVa of female breeders compared with female subordinates or male breeders suggests that emergence from a hypogonadotrophic state in females may involve kisspeptin‐related mechanisms similar to those underlying puberty or seasonal breeding in other species. J. Comp. Neurol. 521: 3003–3029, 2013.

Experimentally decoupling reproductive investment from energy storage to test the functional basis of a life-history trade-off

The ubiquitous life-history trade-off between reproduction and survival has long been hypothesized to reflect underlying energy-allocation trade-offs between reproductive investment and processes related to self-maintenance. Although recent work has questioned whether energy-allocation models provide sufficient explanations for the survival cost of reproduction, direct tests of this hypothesis are rare, especially in wild populations. This hypothesis was tested in a wild population of brown anole lizards (Anolis sagrei) using a two-step experiment. First, stepwise variation in reproductive investment was created using unilateral and bilateral ovariectomy (OVX) along with intact (SHAM) control. Next, this manipulation was decoupled from its downstream effects on energy storage by surgically ablating the abdominal fat stores from half of the females in each reproductive treatment. As predicted, unilateral OVX (intermediate reproductive investment) induced levels of growth, body condition, fat storage and breeding-season survival that were intermediate between the high levels of bilateral OVX (no reproductive investment) and the low levels of SHAM (full reproductive investment). Ablation of abdominal fat bodies had a strong and persistent effect on energy stores, but it did not influence post-breeding survival in any of the three reproductive treatments. This suggests that the energetic savings of reduced reproductive investment do not directly enhance post-breeding survival, with the caveat that only one aspect of energy storage was manipulated and OVX itself had no overall effect on post-breeding survival. This study supports the emerging view that simple energy-allocation models may often be insufficient as explanations for the life-history trade-off between reproduction and survival.

Structural coloration signals condition, parental investment, and circulating hormone levels in Eastern bluebirds ( Sialia sialis )

Many of the brilliant plumage coloration displays of birds function as signals to conspecifics. One species in which the function of plumage ornaments has been assessed is the Eastern bluebird (Sialia sialis). Studies of a population breeding in Alabama (USA) have established that plumage ornaments signal quality, parental investment, and competitive ability in both sexes. Here we tested the additional hypotheses that (1) Eastern bluebird plumage ornamentation signals nest defense behavior in heterospecific competitive interactions and (2) individual variation in plumage ornamentation reflects underlying differences in circulating hormone levels. We also tested the potential for plumage ornaments to signal individual quality and parental investment in a population breeding in Oklahoma (USA). We found that Eastern bluebirds with more ornamented plumage are in better condition, initiate breeding earlier in the season, produce larger clutches, have higher circulating levels of the stress hormone corticosterone, and more ornamented males have lower circulating androgen levels. Plumage coloration was not related to nest defense behavior. Thus, plumage ornamentation may be used by both sexes to assess the physiological condition and parental investment of prospective mates. Experimental manipulations of circulating hormone levels during molt are needed to define the role of hormones in plumage ornamentation.

Temperature Effects During Early Life Stages of the Alligator Snapping Turtle (Macrochelys temminckii)

Abstract Alligator snapping turtle (Macrochelys temminckii) populations have declined across much of the southeastern United States in recent decades, due at least in part to overcollection. Recently, however, legal protection from large-scale harvesting has been granted to the species in all states where it is native, thereby drastically reducing one of the greatest threats to its survival. There is growing interest in captive propagation of alligator snapping turtles for reintroduction where populations have been decimated. In conjunction with one such effort, we analyzed the physiological effects of temperature on embryonic and posthatching development. Results indicate that extreme high and low incubation temperatures negatively affected embryo survival, and high incubation temperatures corresponded with shorter incubation time but also produced smaller hatchlings. The effects of temperature on gonadal differentiation indicated that the upper pivotal temperature was approximately 27.5°C. Posthatching growth was faster at warmer water temperatures, and there was little to no acclimation of metabolic rate to exposure to either incubation or water temperature. We conclude that intermediate (27.5°–28.5°C) incubation temperatures produce a female-biased mixed sex ratio and maximize hatching success and hatchling size while increasing incubation duration only slightly over that at the higher temperatures. In addition, posthatching growth was positively influenced by hatchling body temperature; therefore, warmer water temperatures (∼30°C) decreased the time required to rear turtles to a size suitable for reintroduction.