Thermal tolerance plasticity and dynamics of thermal tolerance in Eublepharis macularius: Implications for future climate-driven heat stress

The intensity and duration of heat waves, as well as average global temperatures, are expected to increase due to climate change. Heat waves can cause physiological stress and reduce fitness in animals. Species can reduce overheating risk through phenotypic plasticity, which allows them to raise their thermal tolerance limits over time. This mechanism could be important for ectotherms whose body temperatures are directly influenced by available environmental temperatures. Geckos are a large, diverse group of ectotherms that vary in their thermal habitats and times of daily activity, which could affect how they physiologically adjust to heat waves. Data on thermal plasticity are scarce for reptiles, with only one study in geckos. Understanding thermal tolerance and plasticity, and their relationship, is essential for understanding how some species are able to adjust or adapt to changing temperatures. In this study, we estimated thermal tolerance and plasticity, and their interaction, in the crepuscular gecko, Eublepharis macularius, a species that is emerging as a model for reptile biology. After estimating basal thermal tolerance for 28 geckos, thermal tolerance was measured for each individual a second time at several time points (3, 6, or 24 h) to determine thermal tolerance plasticity. We found that thermal tolerance plasticity (1) does not depend on the basal thermal tolerance of the organism, (2) was highest after 6 hours from initial heat shock, and (3) was negatively influenced by individual body mass. Our findings contribute to the increasing body of work focused on understanding the influence of biological and environmental factors on thermal tolerance plasticity in organisms and provide phenotypic data to further investigate the molecular basis of thermal tolerance plasticity in organisms.

Methods
In this study, we established the critical thermal maximum (basal upper thermal tolerance), inferred the thermal tolerance plasticity, and assessed their relationship in the gecko, Eublepharis macularius. Thermal tolerance plasticity through heat hardening has been measured as the change in the critical thermal maximum (CTmax) over time. CTmax corresponds to the upper thermal tolerance of an organism, which is the internal body temperature at which the organism loses physiological function and cannot tolerate any higher temperatures. In addition to measuring heat hardening capacity and thermal tolerance within E. macularius, in this work, we also investigate the influence of individual body mass, body size, and sex on heating rates across CTmax experiments.