Net energy availability depends on plasma corticosterone concentrations, food availability, and their interaction. Limited net energy availability requires energy trade-offs between self-maintenance and reproduction. This is important in matrotrophic viviparous animals because they provide large amounts of energy for embryos, as well as self-maintenance, for the extended period of time during gestation. In addition, gravid females may transmit environmental information to the embryos in order to adjust offspring phenotype. We investigated effects of variation in maternal plasma corticosterone concentration and maternal food availability (2 # 2 factorial design) during gestation on offspring phenotype in a matrotrophic viviparous lizard (Pseudemoia entrecasteauxii). Subsequently, we tested preadaptation of offspring phenotype to their postnatal environment by measuring risk-averse behavior and growth rate using reciprocal transplant experiments. We found that maternal net energy availability affected postpartum maternal body condition, offspring snout-vent length, offspring mass, offspring performance ability, and offspring fat reserves. Females treated with corticosterone allocated large amounts of energy to their own body condition, and their embryos allocated more energy to energy reserves than somatic growth. Further, offspring from females in high plasma corticosterone concentration showed compensatory growth. These findings suggest that while females may be selfish when gestation conditions are stressful, the embryos may adjust their phenotype to cope with the postnatal environment.

The evolution of matrotrophy (i.e., direct supply of nutrients by the mother during gestation) may be associated with high maternal energy availability during gestation. However, we lack knowledge about the selective advantages of matrotrophic viviparity (live-bearing) in reptiles. In reptiles, the interaction between body temperature and food intake affect maternal net energy gain. In the present study, we examined the effects of basking and food availability (2 by 2 factorial design) during gestation on offspring phenotype in a matrotrophic viviparous lizard (Pseudemoia entrecasteauxii). Subsequently, we investigated if the maternal effects were context-dependent using offspring growth rate as an indicator of the adaptive significance of matrotrophy. Offspring were exposed either to the same thermal conditions as their mothers experienced or to thermal conditions different from those experienced by their mothers. We provide the first evidence that an interaction between maternal thermal and maternal food conditions during gestation strongly affects offspring phenotype, including date of birth, body size and performance ability, which affect offspring fitness. Offspring growth rate was dependent on offspring thermal conditions, but was not influenced by maternal effects or offspring sex. Matrotrophic viviparity provided gravid females with the means to enhance offspring fitness through greater energetic input to offspring when conditions allowed it (i.e., extended basking opportunity with high food availability). Therefore, we suggest that selective advantages of matrotrophic viviparity in P. entrecasteauxii may be associated with high maternal energy availability during gestation.

Placentotrophy is the nourishment of embryos by resources provided via the placenta during gestation. The magnitude and timing of placental nutrient support during pregnancy are important for embryonic growth, especially in highly placentotrophic animals such as mammals. However, no study has yet investigated how placental organic nutrient support may change during pregnancy in highly placentotrophic viviparous reptiles. Amino acids are essential nutrients for embryonic growth and leucine is a common amino acid. The magnitude and timing of placental leucine transfer may affect embryonic growth and mass and, therefore, offspring phenotype. In this study, female Pseudemoia entrecasteauxii, a highly placentotrophic viviparous skink, were collected throughout gestation. We injected (3)H-leucine into these gravid females and assessed the transfer of (3)H-leucine into maternal compartments (i.e., the blood and the liver), and into embryonic compartments (i.e., the embryo, the yolk, and the amniotic fluid). At either 60 or 120 min post-injection, the radioactivity in each sample was extracted and then counted, and the transfer ratio was calculated. Our results provide direct evidence that circulating maternal leucine passes through the placenta into the embryos in this species. The relative rate of placental leucine transfer did not alter during mid to late gestation. This suggests the steady somatic growth of the embryos during mid-late pregnancy is dependent upon the placental transfer of nutrients rather than yolk stores. This pattern of placental nutrient support may determine offspring body size at birth and, therefore, offspring fitness in P. entrecasteauxii.

Summary1. The evolution of matrotrophy may be related to high resource availability during gestation and consequently increased opportunity for reproductive investment via the placenta. Thus, matrotrophic species have smaller yolk volume than lecithotrophic species. Innate immune response of offspring at birth, which is important in protection against disease and parasites, may be influenced by the degree of reproductive investment of carotenoids, such as b-carotene. However, to date, no study has investigated the effects of maternal carotenoid intake during reproduction on the innate immune responses of offspring in reptiles. 2. Some matrotrophic viviparous (live-bearing) reptiles provide significant support for embryogenesis through both yolk and placenta. These species are excellent models for research into the evolutionary transitions from lecithotrophic reproduction (i.e. supply of resources from the yolks, e.g. birds) to highly matrotrophic reproduction (i.e. direct supply of most resources from the mother, e.g. mammals). 3. We used high-performance liquid chromatography (HPLC) to examine carotenoid concentration in newly ovulated eggs of a matrotrophic viviparous lizard (Pseudemoia entrecasteauxii). Most eggs did not contain any carotenoids, including b-carotene. 4. We then experimentally manipulated maternal b-carotene availability (i.e. supplementation of b-carotene in the diet or no supplementation) during gestation and investigated innate immune response of offspring (i.e. tissue swelling and proportion of monocytes in response to antigenic stimulation) at 2 days after birth. Subsequently, we investigated the effects of postnatal b-carotene intake on innate immune response of offspring using reciprocal transplant experiments in which we manipulated offspring b-carotene availability for 2 weeks after birth. This experimental design also allowed us to assess the persistence of effects of maternal b-carotene effects on innate immune response in offspring. We found that the immune response as measured by the proportion of monocytes was enhanced by high maternal b-carotene intake during gestation, and this effect persisted for 2 weeks after birth. Furthermore, post-natal b-carotene intake enhanced innate immune response. 5. The maternally transferred ß-carotene via the placenta during embryogenesis may be an important physiological adaptation to counter the costs of a reduction in organic substances supplied in the yolk that occurs as matrotrophy evolves.

Interpopulational variation in reproductive costs may affect variation in life history traits including reproductive investment (i.e. clutch mass relative to either maternal body mass or length). While the relationships between reproductive investment and costs of reproduction, especially costs to mobility, have been well studied in squamate reptiles, how these costs relate to investment and explain patterns within and between populations is not always straightforward. In the present study, we examined the relationship between reproductive investment and costs of reproduction (gravid and postpartum sprint speeds and maternal postpartum body condition) in two populations of a viviparous skink, Pseudemoia entrecasteauxii living in different habitat types. We found that costs of reproduction (i.e. impact on gravid and postpartum sprint speeds) depended on the interaction between relative reproductive burden (RRB) and population. There was no link between relative clutch mass (RCM) and maternal sprint speeds. Maternal postpartum body condition was not related to either RRB or RCM for either population. Gravid females living in the open habitat population showed significantly slower sprint speed compared with the same females immediately postparturition, and other gravid females living in a closed habitat population. Such females are likely to experience a higher cost of reproduction in terms of changes in sprint speed as well as exposure to predators and may show a behavioural shift to crypsis in order to compensate for locomotor impairment and to reduce the risk of predation. We suggest that factors which relate to costs of reproduction (i.e. sprint speeds) are complex and may involve multiple factors such as reproductive investment and habitat characteristics.