Kylie A. Robert

Viviparous lizard selects sex of embryos

No one suspected that temperature-dependent sex determination (TSD), whereby the sex of embryos depends on the temperature at which they develop, might occur in viviparous (live-bearing) reptiles, because thermoregulation in the mother results in relatively stable, raised gestation temperatures. But here we show that developing embryos of the actively thermoregulating viviparous skink Eulamprus tympanum are subject to TSD, offering the mother the chance to select the sex of her offspring and a mechanism to help to balance sex ratios in wild populations.

Evolution of senescence in nature: physiological evolution in populations of garter snake with divergent life histories.

Evolutionary theories of aging are linked to life‐history theory in that age‐specific schedules of reproduction and survival determine the trajectory of age‐specific mutation/selection balances across the life span and thus the rate of senescence. This is predicted to manifest at the organismal level in the evolution of energy allocation strategies of investing in somatic maintenance and robust stress responses in less hazardous envirnments in exchange for energy spent on growth and reproduction. Here we report experiments from long‐studied populations of western terrestrial garter snakes (Thamnophis elegans) that reside in low and high extrinsic mortality environments, with evolved long and short life spans, respectively. Laboratory common‐environment colonies of these two ecotypes were tested for a suite of physiological traits after control and stressed gestations. In offspring derived from control and corticosterone‐treated dams, we measured resting metabolism; mitochondrial oxygen consumption, ATP and free radical production rates; and erythrocyte DNA damage and repair ability. We evaluated whether these aging biomarkers mirrored the evolution of life span and whether they were sensitive to stress. Neonates from the long‐lived ecotype (1) were smaller, (2) consumed equal amounts of oxygen when corrected for body mass, (3) had DNA that damaged more readily but repaired more efficiently, and (4) had more efficient mitochondria and more efficient cellular antioxidant defenses than short‐lived snakes. Many ecotype differences were enhanced in offspring derived from stress‐treated dams, which supports the conclusion that nongenetic maternal effects may further impact the cellular stress defenses of offspring. Our findings reveal that physiological evolution underpins reptilian life histories and sheds light on the connectedness between stress response and aging pathways in wild‐dwelling organisms.

The effects of maternal corticosterone levels on offspring behavior in fast- and slow-growth garter snakes (Thamnophis elegans)

During embryonic development, viviparous offspring are exposed to maternally circulating hormones. Maternal stress increases offspring exposure to corticosterone and this hormonal exposure has the potential to influence developmental, morphological and behavioral traits of the resulting offspring. We treated pregnant female garter snakes (Thamnophis elegans) with low levels of corticosterone after determining both natural corticosterone levels in the field and pre-treatment levels upon arrival in the lab. Additional measurements of plasma corticosterone were taken at days 1, 5, and 10 during the 10-day exposure, which occurred during the last third of gestation (of 4-month gestation). These pregnant snakes were from replicate populations of fast- and slow-growth ecotypes occurring in Northern California, with concomitant short and long lifespans. Field corticosterone levels of pregnant females of the slow-growth ecotype were an order of magnitude higher than fast-growth dams. In the laboratory, corticosterone levels increased over the 10 days of corticosterone manipulation for animals of both ecotypes, and reached similar plateaus for both control and treated dams. Despite similar plasma corticosterone levels in treated and control mothers, corticosterone-treated dams produced more stillborn offspring and exhibited higher total reproductive failure than control dams. At one month of age, offspring from fast-growth females had higher plasma corticosterone levels than offspring from slow-growth females, which is opposite the maternal pattern. Offspring from corticosterone-treated mothers, although unaffected in their slither speed, exhibited changes in escape behaviors and morphology that were dependent upon maternal ecotype. Offspring from corticosterone-treated fast-growth females exhibited less anti-predator reversal behavior; offspring from corticosterone-treated slow-growth females exhibited less anti-predator tail lashing behavior.

Testing the 'free radical theory of aging' hypothesis: physiological differences in long-lived and short-lived colubrid snakes

We test the ‘free radical theory of aging’ using six species of colubrid snakes (numerous, widely distributed, non‐venomous snakes of the family Colubridae) that exhibit long (> 15 years) or short (< 10 years) lifespans. Because the ‘rate of living theory’ predicts metabolic rates to be correlated with rates of aging and oxidative damage results from normal metabolic processes we sought to answer whether physiological parameters and locomotor performance (which is a good predictor of survival in juvenile snakes) mirrored the evolution of lifespans in these colubrid snakes. We measured whole animal metabolic rate (oxygen consumption ), locomotor performance, cellular metabolic rate (mitochondrial oxygen consumption), and oxidative stress potential (hydrogen peroxide production by mitochondria). Longer‐lived colubrid snakes have greater locomotor performance and reduced hydrogen peroxide production than short‐lived species, while whole animal metabolic rates and mitochondrial efficiency did not differ with lifespan. We present the first measures testing the ‘free radical theory of aging’ using reptilian species as model organisms. Using reptiles with different lifespans as model organisms should provide greater insight into mechanisms of aging.

Energy consumption by embryos of a viviparous lizard, Eulamprus tympanum, during development.

Energy consumption during development has been measured in many oviparous lizards, but not in viviparous lizards in utero. It has always been assumed that energy consumption by embryos of viviparous lizards during development is similar to that of oviparous species. Estimation of energy consumption of viviparous lizards in vivo are confounded by the possible influence of pregnancy on maternal metabolism. Here we separated maternal and embryonic metabolism in measurements of pregnant Eulamprus tympanum throughout pregnancy. Our data support the hypothesis that the energetic cost of development in viviparous lizards (19.8 kJ g(-1)) is similar to that in oviparous lizards (mean 16.2 kJ g(-1)), at least for a species with a simple placenta. An increase in maternal metabolism of 29% above that for non-pregnant E. tympanum goes to maintain pregnancy, and represents an important component of the reproductive effort in E. tympanum.

Artificial light at night desynchronizes strictly seasonal reproduction in a wild mammal.

Change in day length is an important cue for reproductive activation in seasonally breeding animals to ensure that the timing of greatest maternal investment (e.g. lactation in mammals) coincides with favourable environmental conditions (e.g. peak productivity). However, artificial light at night has the potential to interfere with the perception of such natural cues. Following a 5-year study on two populations of wild marsupial mammals exposed to different night-time levels of anthropogenic light, we show that light pollution in urban environments masks seasonal changes in ambient light cues, suppressing melatonin levels and delaying births in the tammar wallaby. These results highlight a previously unappreciated relationship linking artificial light at night with induced changes in mammalian reproductive physiology, and the potential for larger-scale impacts at the population level.

Influence of Feeding on the Metabolic Rate of the Lizard, Eulamprus tympanum

Abstract We measured the increase in metabolic rate following feeding, or the specific dynamic action (SDA) of feeding, in the lizard Eulamprus tympanum that feeds opportunistically and frequently. The SDA of most reptiles studied to date is pronounced in relation to other vertebrates, but most species for which data are available consume large meals infrequently. We chose to measure SDA in E. tympanum because it is typical of most lizards in that it feeds frequently. The SDA pattern of E. tympanum is typical of other animals, where metabolic rate increases to a peak following feeding then gradually decreases to prefeeding values. Metabolic rate in E. tympanum increases by up to 2.4 times the prefeeding values and remains elevated for up to 48 h. The SDA effect is much less in a lizard that feeds opportunistically and frequently than in sit-and-wait foraging reptiles. The duration of the SDA is greater in E. tympanum than in many other vertebrates and may be related to an ectothermic lifestyle, meal size, or diet composition. On the basis of diet and feeding strategy, E. tympanum is probably typical of lizards that feed frequently, but there are too few data to make detailed comparisons.

Offspring sex varies with maternal investment ability: empirical demonstration based on cross-fostering

Despite decades of interest, adaptive explanations for biased offspring sex ratios in mammals remain contentious, largely because direct tests of the underlying fitness assumptions of adaptive hypotheses are rarely conducted. These tests are complicated by the difficulty of manipulating offspring sex prior to significant maternal investment owing to the biological constraints of viviparity. We test the adaptive advantage of sex allocation through cross-fostering offspring by sex in tammar wallabies. We examine whether offspring sex is correlated with maternal investment ability (i.e. Trivers–Willard hypothesis, TWH). In addition, we test the assumption that maternal investment has a greater influence on the fitness of sons than of daughters. We failed to find statistical support for maternal investment ability influencing a sons weaning success or body size more than a daughters, although this result was probably owing to small sample sizes. In support of the TWH, females that gave birth to a son had higher investment ability (likelihood of weaning an offspring) regardless of the sex of offspring fostered.

Facultative sex allocation in the viviparous lizard Eulamprus tympanum, a species with temperature-dependent sex determination

Females of the Australian scincid lizard Eulamprus tympanum can manipulate the sex of their offspring in response to gender imbalances in the population using temperature-dependent sex determination. Here we show that when adult males are scarce females produced male-biased litters and when adult males were common females produced female-biased litters. The cues used by a female to assess the adult population are not known but presumably depend upon her experience throughout the breeding season. Maternal manipulation of the sex ratio of the offspring in E. tympanum illustrates a selective advantage of temperature-dependent sex determination in a viviparous species.

Milk Composition during Lactation Suggests a Mechanism for Male Biased Allocation of Maternal Resources in the Tammar Wallaby (Macropus eugenii)

Recent research has found empirical evidence in support of the Trivers-Willard Hypothesis that offspring sex allocation is correlated with maternal investment. Tammar wallabies birthing sons have higher investment ability; however a mechanism for sex specific differential allocation of maternal resources in wallabies remains elusive. In metatherians the majority of maternal investment is during lactation. To examine if differential allocation occurs during lactation, we measured total milk protein, lipid and carbohydrates, from mothers with male and female pouch young, during phase 2B (100–215 days post partum) and phase 3 (215–360 days post partum) of lactation. Mothers of sons allocated significantly higher levels of protein than mothers of daughters during phase 2B of lactation, however no sex specific difference in maternal allocation was found for lipids, carbohydrates, or any milk component during phase 3 of lactation. We were unable to measure milk production to establish any differences in the amount of milk allocated. However, with the production of more milk comes a dilution effect on milk components. Given that we find no apparent dilution of milk components may suggest equality in milk production. Offspring body weight at 14 months of age was related to protein allocation during phase 2B of lactation, providing a maternal mechanism for differential allocation with fitness consequences. We believe collection of earlier phase 2A (0–100 days post partum) milk may yield important results given that differential investment in metatherians may be most apparent early in lactation, prior to any significant maternal investment, when a decision on termination of investment can be made with very little energetic loss to the mother. Interestingly, small mothers did not birth sons and better maternal condition was associated with raising sons. These data are in support of TWH and demonstrate a potential mechanism through which condition dependent and sex specific maternal investment may occur.