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Animals spend a considerable proportion of their life span at rest. However, resting status has often been overlooked when investigating how species respond to environmental conditions. This may induce a large bias in understanding the local adaptation of species across environmental gradients and their vulnerability to potential environmental change. Here, we conducted an empirical study on montane agamid lizards, combined with mechanistic modeling, to compare elevational variations in body temperature and metabolisms (cumulative digestion and maintenance cost) between resting and active status. Our study on three populations of an agamid lizard along an elevational gradient revealed a trend of decreasing body temperature toward higher elevations, the main contributor of which was resting status of the lizards. Using population-specific reaction norms, we predicted greater elevational variation in hourly and cumulative digestion for resting lizards than for active lizards. Climate-change impacts, estimated as the change in cumulative digestion, also show greater elevational variation when resting status is factored into the analysis. Further, our global analysis of 98 agamid species revealed that in about half of their combined distributional range, the contribution of resting status in determining the elevational variation in cumulative digestion and maintenance cost of lizards was greater than the contribution made by a lizard's active status. Our study highlights the importance of considering resting status when investigating how species respond to environmental conditions, especially for those distributed over tropical and subtropical mountain areas., (© 2024 The Ecological Society of America.)

Survival of the immobile embryo in response to rising temperature is important to determine a species' vulnerability to climate change. However, the collective effects of 2 key thermal characteristics associated with climate change (i.e., rising average temperature and acute heat events) on embryonic survival remain largely unexplored. We used empirical measurements and niche modeling to investigate how chronic and acute heat stress independently and collectively influence the embryonic survival of lizards across latitudes. We collected and bred lizards from 5 latitudes and incubated their eggs across a range of temperatures to quantify population-specific responses to chronic and acute heat stress. Using an embryonic development model parameterized with measured embryonic heat tolerances, we further identified a collective impact of embryonic chronic and acute heat tolerances on embryonic survival. We also incorporated embryonic chronic and acute heat tolerance in hybrid species distribution models to determine species' range shifts under climate change. Embryos' tolerance of chronic heat (T-chronic) remained consistent across latitudes, whereas their tolerance of acute heat (T-acute) was higher at high latitudes than at low latitudes. Tolerance of acute heat exerted a more pronounced influence than tolerance of chronic heat. In species distribution models, climate change led to the most significant habitat loss for each population and species in its low-latitude distribution. Consequently, habitat for populations across all latitudes will shift toward high latitudes. Our study also highlights the importance of considering embryonic survival under chronic and acute heat stresses to predict species' vulnerability to climate change., (© 2024 The Authors. Conservation Biology published by Wiley Periodicals LLC on behalf of Society for Conservation Biology.)

Environmental temperatures are increasing worldwide, threatening desert ectotherms already living at their thermal limits. Organisms with flexible thermoregulatory behaviours may be able to mitigate the effects of extreme temperatures by moving among microhabitats, yet little work has tracked movement patterns of desert ectotherms in the wild over diurnal scales or compared behaviour among seasons. Here, we used camera traps to track the thermoregulatory behaviour and microhabitat choices of 30 desert lizards (Messalina bahaldini) in custom, outdoor arenas that provided access to open, rock, and bush microhabitats. We found that in the summer, lizards preferred to move to the shaded microhabitats and remain there under warmer conditions. During winter, however, lizards' activity was not related to temperature, and lizards mostly chose to remain in the open habitat. Interestingly, in both seasons, lizards tended to remain in their current microhabitat and moved infrequently between certain combinations of microhabitats. Our study shows that thermoregulation (shade-seeking behaviour) is a major factor during summer, helping lizards to avoid extreme temperatures, but not during winter, and shows a novel effect of current microhabitat on movement, suggesting that other biotic or abiotic factors may also drive microhabitat choice. Understanding the complex factors at play in microhabitat choice is critical for developing conservation programs that effectively mitigate the negative impacts of climate change on desert animals., Competing Interests: Declaration of competing interest There are no competing interests among all authors for this manuscript., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Local adaptation is critical in speciation and evolution, yet comprehensive studies on proximate and ultimate causes of local adaptation are generally scarce. Here, we integrated field ecological experiments, genome sequencing, and genetic verification to demonstrate both driving forces and molecular mechanisms governing local adaptation of body coloration in a lizard from the Qinghai-Tibet Plateau. We found dark lizards from the cold meadow population had lower spectrum reflectance but higher melanin contents than light counterparts from the warm dune population. Additionally, the colorations of both dark and light lizards facilitated the camouflage and thermoregulation in their respective microhabitat simultaneously. More importantly, by genome resequencing analysis, we detected a novel mutation in Tyrp1 that underpinned this color adaptation. The allele frequencies at the site of SNP 459# in the gene of Tyrp1 are 22.22% G/C and 77.78% C/C in dark lizards and 100% G/G in light lizards. Model-predicted structure and catalytic activity showed that this mutation increased structure flexibility and catalytic activity in enzyme TYRP1, and thereby facilitated the generation of eumelanin in dark lizards. The function of the mutation in Tyrp1 was further verified by more melanin contents and darker coloration detected in the zebrafish injected with the genotype of Tyrp1 from dark lizards. Therefore, our study demonstrates that a novel mutation of a major melanin-generating gene underpins skin color variation co-selected by camouflage and thermoregulation in a lizard. The resulting strong selection may reinforce adaptive genetic divergence and enable the persistence of adjacent populations with distinct body coloration., Competing Interests: Conflict of interest statement. The authors declare no conflict of interest., (© The Author(s) 2024. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

After drawing comparisons between the reaction pathways of cytochrome c oxidase (CcO, Complex 4) and the preceding complex cytochrome bc 1 (Complex 3), both being proton pumping complexes along the electron transport chain, we provide an analysis of the reaction pathways in bacterial ba 3 class CcO, comparing spectroscopic results and kinetics observations with results from DFT calculations. For an important arc of the catalytic cycle in CcO, we can trace the energy pathways for the chemical protons and show how these pathways drive proton pumping of the vectorial protons. We then explore the proton loading network above the Fe heme a 3 -Cu B catalytic center, showing how protons are loaded in and then released by combining DFT-based reaction energies with molecular dynamics simulations over states of that cycle. We also propose some additional reaction pathways for the chemical and vector protons based on our recent work with spectroscopic support., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Noodleman, Götz, Han Du and Hunsicker-Wang.)

Worldwide habitat loss, land-use changes, and climate change threaten biodiversity, and we urgently need models that predict the combined impacts of these threats on organisms. Current models, however, overlook microhabitat diversity within landscapes and so do not accurately inform conservation efforts, particularly for ectotherms. Here, we built and field-parameterized a model to examine the effects of habitat loss and climate change on activity and microhabitat selection by a diurnal desert lizard. Our model predicted that lizards in rock-free areas would reduce summer activity levels (e.g. foraging, basking) and that future warming will gradually decrease summer activity in rocky areas, as even large rocks become thermally stressful. Warmer winters will enable more activity but will require bushes and small rocks as shade retreats. Hence, microhabitats that may seem unimportant today will become important under climate change. Modelling frameworks should consider the microhabitat requirements of organisms to improve conservation outcomes., (© 2023 The Authors. Global Change Biology published by John Wiley & Sons Ltd.)

Climate change often includes increases in the occurrence of extreme environmental events. Among these, heatwaves affect the pace of life and performance of wildlife, particularly ectothermic animals, owing to their low thermoregulatory abilities. However, the underlying mechanisms by which this occurs remain unclear. Evidence shows that heatwaves alter the redox balance of ectotherms, and oxidative stress is a major mediator of life-history trade-offs. Therefore, oxidative stress may mediate the effect of extreme thermal conditions on the life histories of ectotherms. To test this hypothesis, a 2 × 2 experiment was conducted to manipulate the redox balance (through a mitochondrial uncoupler that alleviates oxidative stress) of the desert toad-headed agama ( Phrynocephalus przewalskii ) exposed to heatwave conditions. We recorded lizard growth and survival rates and quantified their redox and immune statuses. In control lizards (unmanipulated redox balance), heatwave conditions decreased growth and survival and induced oxidative damage and immune responses. By contrast, lizards with alleviated oxidative stress showed close-to-normal growth, survival, and immune status when challenged with heatwaves. These results provide mechanistic insight into the role of oxidative stress in mediating the effects of extreme temperatures on ectothermic vertebrates, which may have major eco-evolutionary implications.

Higher temperatures enhance ectothermic metabolism and development, which can reduce individual health and life expectancy, and therefore increase their vulnerability to climate warming. However, the mechanistic causes and consequences of such a temperature-driven impact remain unclear. Our study aimed to address two questions: (1) does climate warming alter early-life growth and physiology, and, if so, what are the associated carry-over effects in terms of reduced survival, increased oxidative stress and telomere shortening? (2) can oxidative stress and telomere dynamics at early life stages predict the effect of climate warming on individual survival? To answer these questions, we conducted a longitudinal experiment under semi-natural conditions where we exposed multiocellated racerunner (Eremias multiocellata) to warming conditions from juvenile to adult stages. We found that exposure to climate warming enhanced growth rates, induced oxidative stress, and shortened telomere length of juvenile lizards. Warming conditions did not induce carry-over effects in terms of altered growth rate or physiology but resulted in increased mortality risk in the later life. Intriguingly, telomere shortening in young individuals was associated with mortality risk later in life. This study improves our mechanistic understanding of how global warming impacts on ectotherms' life-history traits, which encourages the inclusion of physiological information in assessing species vulnerability to climate change., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

A range of abiotic parameters within a reptile nest influence the viability and attributes (including sex, behaviour and body size) of hatchlings that emerge from that nest. As a result of that sensitivity, a reproducing female can manipulate the phenotypic attributes of her offspring by laying her eggs at times and in places that provide specific conditions. Nesting reptiles shift their behaviour in terms of timing of oviposition, nest location and depth of eggs beneath the soil surface across spatial and temporal gradients. Those maternal manipulations affect mean values and variances of both temperature and soil moisture, and may modify the vulnerability of embryos to threats such as predation and parasitism. By altering thermal and hydric conditions in reptile nests, climate change has the potential to dramatically modify the developmental trajectories and survival rates of embryos, and the phenotypes of hatchlings. Reproducing females buffer such effects by modifying the timing, location and structure of nests in ways that enhance offspring viability. Nonetheless, our understanding of nesting behaviours in response to climate change remains limited in reptiles. Priority topics for future studies include documenting climate-induced changes in the nest environment, the degree to which maternal behavioural shifts can mitigate climate-related deleterious impacts on offspring development, and ecological and evolutionary consequences of maternal nesting responses to climate change. This article is part of the theme issue 'The evolutionary ecology of nests: a cross-taxon approach'.

Climate warming can substantially impact embryonic development and juvenile growth in oviparous species. Estimating the overall impacts of climate warming on oviparous reproduction is difficult because egg-laying events happen throughout the reproductive season. Successful egg laying requires the completion of embryonic development as well as hatching timing conducive to offspring survival and energy accumulation. We propose a new metric-egg-laying opportunity (EO)-to estimate the annual hours during which a clutch of freshly laid eggs yields surviving offspring that store sufficient energy for overwintering. We estimated the EO within the distribution of a model species, Sceloporus undulatus, under recent climate condition and a climate-warming scenario by combining microclimate data, developmental functions, and biophysical models. We predicted that EO will decline as the climate warms at 74.8% of 11,407 sites. Decreasing hatching success and offspring energy accounted for more lost EO hours (72.6% and 72.9%) than the occurrence of offspring heat stress (59.9%). Nesting deeper (at a depth of 12 cm) may be a more effective behavioral adjustment for retaining EO than using shadier (50% shade) nests because the former fully mitigated the decline of EO under the considered warming scenario at more sites (66.1%) than the latter (28.3%). We advocate for the use of EO in predicting the impacts of climate warming on oviparous animals because it encapsulates the integrative impacts of climate warming on all stages of reproductive life history., (© 2023 Society for Conservation Biology.)

Ongoing climate change has profoundly affected global biodiversity, but its impacts on populations across elevations remain understudied. Using mechanistic niche models incorporating species traits, we predicted ecophysiological responses (activity times, oxygen consumption and evaporative water loss) for lizard populations at high-elevation (<3600 m asl) and extra-high-elevation (≥3600 m asl) under recent (1970-2000) and future (2081-2100) climates. Compared with their high-elevation counterparts, lizards from extra-high-elevation are predicted to experience a greater increase in activity time and oxygen consumption. By integrating these ecophysiological responses into hybrid species distribution models (HSDMs), we were able to make the following predictions under two warming scenarios (SSP1-2.6, SSP5-8.5). By 2081-2100, we predict that lizards at both high- and extra-high-elevation will shift upslope; lizards at extra-high-elevation will gain more and lose less habitat than will their high-elevation congeners. We therefore advocate the conservation of high-elevation species in the context of climate change, especially for those populations living close to their lower elevational range limits. In addition, by comparing the results from HSDMs and traditional species distribution models, we highlight the importance of considering intraspecific variation and local adaptation in physiological traits along elevational gradients when forecasting species' future distributions under climate change., (© 2023 John Wiley & Sons Ltd.)

Thickness reduction or loss of the calcareous eggshell is one of major phenotypic changes in the transition from oviparity to viviparity. Whether the reduction of eggshells in viviparous squamates is associated with specific gene losses is unknown. Taking advantage of a newly generated high-quality genome of the viviparous Chinese crocodile lizard (Shinisaurus crocodilurus), we found that ovocleidin-17 gene (OC-17), which encodes an eggshell matrix protein that is essential for calcium deposition in eggshells, is not intact in the crocodile lizard genome. Only OC-17 transcript fragments were found in the oviduct transcriptome, and no OC-17 peptides were identified in the eggshell proteome of crocodile lizards. In contrast, OC-17 was present in the eggshells of the oviparous Mongolia racerunner (Eremias argus). Although the loss of OC-17 is not common in viviparous species, viviparous squamates show fewer intact eggshell-specific proteins than oviparous squamates. Our study implies that functional loss of eggshell-matrix protein genes may be involved in the reduction of eggshells during the transition from oviparity to viviparity in the crocodile lizard., (© 2022 European Society for Evolutionary Biology.)

Microhabitats provide ecological and physiological benefits to animals, sheltering them from predation and extreme temperatures and offering an additional supply of water and food. However, most studies have assumed no energetic costs of searching for microhabitats or moving between them, or considered how the availability of microhabitats may affect the energy reserves of animals and how such effects may differ between seasons. To fill these gaps, we studied how the body condition of lizards is affected by microhabitat availability in the extreme environment of the Judean Desert. In particular, we quantified how vegetation and rock cover in the vicinity of these lizards affect their body condition during summer and winter. First, we used aerial imagery to map the vegetation/rock cover at two study sites. Next, we collected 68 adult lizards and examined how their body condition varies across seasons and availability of vegetation and rock cover. In addition, we examined how vegetation and rock cover may differ in their effective distance (i.e., the distance that best explains body condition of lizards). We found that lizards body condition was better if they were collected closer to a higher availability of vegetation or rocks. However, while close proximity (within 10 m) was the best predictor for the positive effect of rocks, a greater distance (up to 90 m) was the best predictor for the effect of the vegetation cover. Moreover, the positive effect of vegetation was 12-fold higher than the effect of rocks. Interestingly, although the lizards' body condition during winter was poorer than during summer, the positive effects of rock and vegetation cover remained constant between the seasons. This similarity of benefits across seasons suggests that shaded microhabitats have important additional ecological roles regardless of climate, and that they may provide thermoregulatory benefits in winter too. We also found a synergic effect of vegetation and rock cover on the lizards' body condition, suggesting that their roles are complementary rather than overlapping. Our research has revealed the importance of shade- and shelter-providing microhabitats in both summer and winter. We suggest that proximity to microhabitat diversity may contribute to better body condition in lizards or, alternatively, facilitates competition and attracts lizards with better body condition. Comprehending the complex interactions between animals and different microhabitats is critical for developing better conservation plans, understanding the risks of climate change, and suggesting mitigation strategies., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Society for Integrative and Comparative Biology.)

Adaptations of post-hatching animals have attracted far more study than have embryonic responses to environmental challenges, but recent research suggests that we have underestimated the complexity and flexibility of embryos. We advocate a dynamic view of embryos as organisms capable of responding - on both ecological and evolutionary timescales - to their developmental environments. By viewing embryos in this way, rather than assuming an inability of pre-hatching stages to adapt and respond, we can broaden the ontogenetic breadth of evolutionary and ecological research. Both biotic and abiotic factors affect embryogenesis, and embryos exhibit a broad range of behavioural and physiological responses that enable them to deal with changes in their developmental environments in the course of interactions with their parents, with other embryos, with predators, and with the physical environment. Such plasticity may profoundly affect offspring phenotypes and fitness, and in turn influence the temporal and spatial dynamics of populations and communities. Future research in this field could benefit from an integrated framework that combines multiple approaches (field investigations, manipulative experiments, ecological modelling) to clarify the mechanisms and consequences of embryonic adaptations and plasticity., (© 2022 Cambridge Philosophical Society.)

Mössbauer isomer shift and quadrupole splitting properties have been calculated using the OLYP-D3(BJ) density functional method on previously obtained (W.-G. Han Du, et al., Inorg Chem. 2020, 59, 8906-8915) geometry optimized Fe a3 3+ -H 2 O-Cu B 2+ dinuclear center (DNC) clusters of the resting oxidized (O state) "as-isolated" cytochrome c oxidase (CcO). The calculated results are highly consistent with the available experimental observations. The calculations have also shown that the structural heterogeneities of the O state DNCs implicated by the Mössbauer experiments are likely consequences of various factors, particularly the variable positions of the central H 2 O molecule between the Fe a3 3+ and Cu B 2+ sites in different DNCs, whether or not this central H 2 O molecule has H-bonding interaction with another H 2 O molecule, the different spin states having similar energies for the Fe a3 3+ sites, and whether the Fe a3 3+ and Cu B 2+ sites are ferromagnetically or antiferromagnetically spin-coupled., (© 2022 The Authors. ChemPhysChem published by Wiley-VCH GmbH.)

The purging of deleterious alleles has been hypothesized to mitigate inbreeding depression, but its effectiveness in endangered species remains debatable. To understand how deleterious alleles are purged during population contractions, we analyzed genomes of the endangered Chinese crocodile lizard (Shinisaurus crocodilurus), which is the only surviving species of its family and currently isolated into small populations. Population genomic analyses revealed four genetically distinct conservation units and sharp declines in both effective population size and genetic diversity. By comparing the relative genetic load across populations and conducting genomic simulations, we discovered that seriously deleterious alleles were effectively purged during population contractions in this relict species, although inbreeding generally enhanced the genetic burden. However, despite with the initial purging, our simulations also predicted that seriously deleterious alleles will gradually accumulate under prolonged bottlenecking. Therefore, we emphasize the importance of maintaining a minimum population capacity and increasing the functional genetic diversity in conservation efforts to preserve populations of the crocodile lizard and other endangered species., (© The Author(s) 2021. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.)

Temperature sensing is essential for the survival of living organisms. Some reptile embryos can reposition themselves within the egg to seek optimal temperatures, but the molecular sensors involved in this temperature detection remain unknown. Here, we show that such thermotaxic behavior is directly determined by the activation of two heat-sensitive ion channels of the turtle: the transient receptor potential ankyrin 1 (MrTRPA1) and transient receptor potential vanilloid-1 (MrTRPV1). These two TRP channels were found to exhibit distinctive distributions among turtle dorsal root ganglion (DRG) neurons. Additionally, our laser irradiation assays illustrated that the heat activation thresholds of MrTRPA1 and MrTRPV1 are consistent with the mild (28-33°C) and noxious (>33°C) heat determined by behavioral tests, respectively. Further pharmacological studies have demonstrated that ligand-induced intervention of MrTRPA1 or MrTRPV1 is sufficient to mimic heat stimuli or block temperature signaling, causing changes in embryo movement. These findings indicate that the initiation of thermotaxic response in turtle embryos relies on a delicate functional balance between the heat activation of MrTRPA1 and MrTRPV1. Our study reveals, for the first time, a unique molecular mechanism underlying thermal detection: the two TRP channels act as a physiological tandem to control the thermotaxic behavior of turtle embryos., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

While the effects of incubation environment on embryonic development and offspring traits have been extensively studied in oviparous vertebrates, studies into how genetic inheritance (population origin), maternal effects, and incubation environment interact to produce varying phenotypes, are rare. To elucidate the interactive role of those three factors during incubation in shaping offspring phenotypes through hydric conditions, we conducted a fully factorial experiment [arid and semiarid populations × maternal dry and wet treatments (MDT and MWT) × embryonic dry and wet treatments (EDT and EWT)] with a desert-dwelling lacertid lizard (Eremias argus). Female lizards in dry conditions produced larger clutch sizes but smaller eggs. The incubation period and hatching success were significantly affected by embryonic but not by maternal moisture treatments. Eggs in the EDT hatched later than those in the EWT in both arid and semiarid populations. Hatching success was lower in EDT than in EWT in the semiarid population, but not in the arid population. Hatchlings from the EDT had a slower post-hatch increase in body mass than those from the EWT. EDT would decrease the survival rates of hatchlings in the semiarid population only. In addition, structural equation models revealed that population had a stronger effect on embryonic and offspring survival than maternal and embryonic moisture. Our study demonstrates locally adaptive strategies of drought resistance at multiple life-history stages in lizard populations from diverse hydric habitats and highlights the importance of genetic factors in determining embryonic drought resistance in oviparous lizards.

As postulated by life-history theory, not all life-history traits can be maximized simultaneously. In ectothermic animals, climate warming is predicted to increase growth rates, but at a cost to overall life span. Maternal effects are expected to mediate this life-history trade-off, but such effects have not yet been explicitly elucidated. To understand maternal effects on the life-history responses to climate warming in lizard offspring, we conducted a manipulative field experiment on a desert-dwelling viviparous lacertid lizard Eremias multiocellata, using open-top chambers in a factorial design (maternal warm climate and maternal present climate treatments × offspring warm climate and offspring present climate treatments). We found that the maternal warm climate treatment had little impact on the physiological and life-history traits of adult females (i.e. metabolic rate, reproductive output, growth and survival). However, the offspring warm climate treatment significantly affected offspring growth, and both maternal and offspring warm climate treatments interacted to affect offspring survival. Offspring from the warm climate treatment grew faster than those from the present climate treatment. However, the offspring warm climate treatment significantly decreased the survival rate of offspring from maternal present climate treatment, but not for those from the maternal warm climate treatment. Our study demonstrates that maternal effects mediate the trade-off between growth and survival of offspring lizards, allowing them to grow fast without a concurrent cost of low survival rate (short life span). These findings stress the importance of adaptive maternal effects in buffering the impact of climate warming on organisms, which may help us to accurately predict the vulnerability of populations and species to future warming climates., (© 2021 British Ecological Society.)

Identifying intrinsic and extrinsic sources of variation in life history traits among populations has been well-studied at the post-embryonic stage but rarely at the embryonic stage. To reveal these sources of variation in the developmental success of embryos, we measured the physical characteristics of nest environments and conducted reciprocal egg-swap experiments in two populations of the toad-headed agamid lizard (Phrynocephalus przewalskii), isolated from each other by a mountain range. We determined the effects of population origin and nest environment on embryonic and offspring traits related to developmental success, including incubation period, hatching success, and offspring growth and survival. Females from the northern population constructed deeper nests that were colder and wetter than those from the southern population. Northern embryos had higher hatching success than the southern embryos when incubated at the northern nest environment, but not when they were incubated at the southern nest environment. The southern hatchlings grew faster than the northern hatchlings when incubated at the southern nest environment, but not after incubation at the northern nest environment. These phenomena likely reflect local adaptation of embryonic development to their nest environments among populations in lizards. In addition, the southern hatchlings had higher survivorship than the northern hatchlings regardless of nest environment, suggesting the southern population has evolved a superior phenotype at the hatchling stage to maximize its fitness.

Montane reptiles are predicted to move to higher elevations in response to climate warming. However, whether upwards-shifting reptiles will be physiologically constrained by hypoxia at higher elevations remains unknown. We investigated the effects of hypoxic conditions on preferred body temperatures (T pref ) and thermal tolerance capacity of a montane lizard ( Phrynocephalus vlangalii ) from two populations on the Qinghai-Tibet Plateau. Lizards from 2600 m a.s.l. were exposed to O 2 levels mimicking those at 2600 m (control) and 3600 m (hypoxia treatment). Lizards from 3600 m a.s.l. were exposed to O 2 levels mimicking those at 3600 m (control) and 4600 m (hypoxia treatment). The T pref did not differ between the control and hypoxia treatments in lizards from 2600 m. However, lizards from 3600 m selected lower body temperatures when exposed to the hypoxia treatment mimicking the O 2 level at 4600 m. Additionally, the hypoxia treatment induced lower critical thermal minimum (CT min ) in lizards from both populations, but did not affect the critical thermal maximum (CT max ) in either population. Our results imply that upwards-shifting reptiles may be constrained by hypoxia if a decrease in T pref reduces thermally dependent fitness traits, despite no observed effect on their heat tolerance.

After a general introduction to the features and mechanisms of cytochrome c oxidases (CcOs) in mitochondria and aerobic bacteria, we present DFT calculated physical and spectroscopic properties for the catalytic reaction cycle compared with experimental observations in bacterial ba3 type CcO, also with comparisons/contrasts to aa3 type CcOs. The Dinuclear Complex (DNC) is the active catalytic reaction center, containing a heme a3 Fe center and a near lying Cu center (called CuB) where by successive reduction and protonation, molecular O2 is transformed to two H2O molecules, and protons are pumped from an inner region across the membrane to an outer region by transit through the CcO integral membrane protein. Structures, energies and vibrational frequencies for Fe-O and O-O modes are calculated by DFT over the catalytic cycle. The calculated DFT frequencies in the DNC of CcO are compared with measured frequencies from Resonance Raman spectroscopy to clarify the composition, geometry, and electronic structures of different intermediates through the reaction cycle, and to trace reaction pathways. X-ray structures of the resting oxidized state are analyzed with reference to the known experimental reaction chemistry and using DFT calculated structures in fitting observed electron density maps. Our calculations lead to a new proposed reaction pathway for coupling the PR → F → OH (ferryl-oxo → ferric-hydroxo) pathway to proton pumping by a water shift mechanism. Through this arc of the catalytic cycle, major shifts in pKa's of the special tyrosine and a histidine near the upper water pool activate proton transfer. Additional mechanisms for proton pumping are explored, and the role of the CuB+ (cuprous state) in controlling access to the dinuclear reaction site is proposed.

Understanding the factors that may affect behavioural thermoregulation of endangered reptiles is important for their conservation because thermoregulation determines body temperatures and in turn physiological functions of these ectotherms. Here we measured seasonal variation in operative environmental temperature (T e ), body temperature (T b ), and microhabitat use of endangered crocodile lizards (Shinisaurus crocodilurus) from a captive population, within open and shaded enclosures, to understand how they respond to thermally challenging environments. T e was higher in open enclosures than in shaded enclosures. The T b of lizards differed between the open and shaded enclosures in summer and autumn, but not in spring. In summer, crocodile lizards stayed in the water to avoid overheating, whereas in autumn, crocodile lizards perched on branches seeking optimal thermal environments. Crocodile lizards showed higher thermoregulatory effectiveness in open enclosures (with low thermal quality) than in shaded enclosures. Our study suggests that the crocodile lizard is capable of behavioural thermoregulation via microhabitat selection, although overall, it is not an effective thermoregulator. Therefore, maintaining diverse thermal environments in natural habitats for behavioural thermoregulation is an essential measure to conserve this endangered species both in the field and captivity., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Although the dinuclear center (DNC) of the resting oxidized "as-isolated" cytochrome c oxidase (C c O) is not a catalytically active state, its detailed structure, especially the nature of the bridging species between the Fe a3 3+ and Cu B 2+ metal sites, is still both relevant and unsolved. Recent crystallographic work has shown an extended electron density for a peroxide type dioxygen species (O1-O2) bridging the Fe a3 and Cu B centers. In this paper, our density functional theory (DFT) calculations show that the observed peroxide type electron density between the two metal centers is most likely a mistaken analysis due to overlap of the electron density of a water molecule located at different positions between apparent O1 and O2 sites in DNCs of different C c O molecules with almost the same energy. Because the diffraction pattern and the resulting electron density map represent the effective long-range order averaged over many molecules and unit cells in the X-ray structure, this averaging can lead to an apparent observed superposition of different water positions between the Fe a3 3+ and Cu B 2+ metal sites.

Captivity is an important measure for conservation of an endangered species, and it is becoming a hot topic in conservation biology, which integrates gut microbiota and endangered species management in captivity. As an ancient reptile, the crocodile lizard ( Shinisaurus crocodilurus ) is facing extreme danger of extinction, resulting in great significance to species conservation in the reserve. Thus, it is critical to understand the differences in gut microbiota composition between captive and wild populations, as it could provide fundamental information for conservative management of crocodile lizards. Here, fecal samples of crocodile lizards were collected from two wild and one captive populations with different ages (i.e., juveniles and adults) and were analyzed for microbiota composition by 16S ribosomal RNA (rRNA) gene amplicon sequencing. This study showed that the lizard gut microbiota was mainly composed of Firmicutes and Proteobacteria. The gut microbiota composition of crocodile lizard did not differ between juveniles and adults, as well as between two wild populations. Interestingly, captivity increased community richness and influenced community structures of gut microbiota in crocodile lizards, compared with wild congeners. This was indicated by higher abundances of the genera Epulopiscium and Glutamicibacter . These increases might be induced by complex integration of simple food resources or human contact in captivity. The gut microbiota functions of crocodile lizards are primarily enriched in metabolism, environmental information processing, genetic information processing, and cellular processes based on the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. This study provides fundamental information about the gut microbiota of crocodile lizards in wild and captive populations. In the future, exploring the relationship among diet, gut microbiota, and host health is necessary for providing animal conservation strategies., (Copyright © 2020 Tang, Liang, Yang, Wang, Chen, Du, Li and Sun.)

Natural nests of egg-laying birds and reptiles exhibit substantial thermal variation, at a range of spatial and temporal scales. Rates and trajectories of embryonic development are highly sensitive to temperature, favouring an ability of embryos to respond adaptively (i.e. match their developmental biology to local thermal regimes). Spatially, thermal variation can be significant within a single nest (top to bottom), among adjacent nests (as a function of shading, nest depth etc.), across populations that inhabit areas with different weather conditions, and across species that differ in climates occupied and/or nest characteristics. Thermal regimes also vary temporally, in ways that generate differences among nests within a single population (e.g. due to seasonal timing of laying), among populations and across species. Anthropogenic activities (e.g. habitat clearing, climate change) add to this spatial and temporal diversity in thermal regimes. We review published literature on embryonic adaptations to spatio-temporal heterogeneity in nest temperatures. Although relatively few taxa have been studied in detail, and proximate mechanisms remain unclear, our review identifies many cases in which natural selection appears to have fine-tuned embryogenesis to match local thermal regimes. Developmental rates have been reported to differ between uppermost versus lower eggs within a single nest, between eggs laid early versus late in the season, and between populations from cooler versus warmer climates. We identify gaps in our understanding of thermal adaptations of early (embryonic) phases of the life history, and suggest fruitful opportunities for future research.

Sessile organisms with thermally sensitive developmental trajectories are at high risk from climate change. For example, oviparous reptiles with temperature-dependent sex determination (TSD) may experience strong (potentially disastrous) shifts in offspring sex ratio if reproducing females are unable to predict incubation conditions at the time of oviposition. How then have TSD reptile taxa persisted over previous periods of extreme climatic conditions? An ability of embryos to move within the egg to select optimal thermal regimes could buffer ambient extremes, but the feasibility of behavioral thermoregulation by embryos has come under strong challenge. To test this idea, we measured thermal gradients within eggs in semi-natural nests of a freshwater turtle species with TSD, manipulated embryonic thermoregulatory ability, and modeled the effects of embryonic thermoregulation on offspring sex ratios. Behavioral thermoregulation by embryos accelerated development and influenced offspring sex ratio, expanding the range of ambient conditions under which nests produce equal numbers of male and female offspring. Model projections suggest that sex ratio shifts induced by global warming will be buffered by the ability of embryos to influence their sexual destiny via behavioral thermoregulation., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Parental effects may produce adaptive or maladaptive plasticity that either facilitates persistence or increases the extinction risk of species and populations in a changing climate. However, empirical evidence of transgenerational adaptive plastic responses to climate change is still scarce. Here we conducted thermal manipulation experiments with a factorial design in a Chinese lacertid lizard ( Takydromus septentrionalis ) to identify the fitness consequences of parental effects in response to climate warming. Compared to present climate conditions, a simulated warming climate significantly advanced the timing of oviposition, depressed the immune capability of post-partum females, and decreased the hatching success of embryos, but did not affect female reproductive output (clutch size and egg mass). These results indicate that maternal warming negatively affects female health, and embryonic hatchability. More interestingly, we found that offspring from parents exposed to warming environments survived well under a simulated warming climate, but not under a present climate scenario. Accordingly, our study demonstrates anticipatory parental effects in response to a warming climate in an ectothermic vertebrate. However, the fitness consequences of this parental effect will depend on future climate change scenarios., Competing Interests: Research was performed under approvals from the Animal Ethics Committee at the Institute of Zoology, Chinese Academy of Sciences (IOZ14001).The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Background: Ectothermic animals living in cold (high latitude or high elevation) regions are predicted to grow slower due to limited thermal opportunities for activity and food resources than those living in warm regions. However, the Qinghai toad-headed lizards (Phrynocephalus vlangalii) grow faster and reach a larger adult size at a high-elevation site than at a low-elevation site. In this study, we aimed to identify the genetic and environmental causes of this between-population difference in growth rate by conducting mark-recapture and common garden experiments on juvenile growth rate, and investigating the thermal environment, lizard body temperature, potential prey availability at the two elevation sites., Results: Compared with low-elevation individuals, high-elevation juvenile lizards had higher growth rates in the field, but grew at similar rates in the laboratory. High-elevation lizards had higher active body temperatures than low-elevation lizards despite similar air temperatures in the period of field investigation. The high-elevation site had relatively more and larger preys than the low-elevation site., Conclusions: Inter-population difference in growth rate of P. vlangalii may primarily result from developmental plasticity in response to the difference in environmental resources, rather than genetic differentiation. The higher growth rate of high-elevation lizards is likely associated with higher potential food availability and higher active body temperatures.

How temperature influences development has direct relevance to ascertaining the impact of climate change on natural populations. Reptiles have served as empirical models for understanding how the environment experienced by embryos can influence phenotypic variation, including sex ratio, phenology and survival. Such an understanding has important implications for basic eco-evolutionary theory and conservation efforts worldwide. While there is a burgeoning empirical literature of experimental manipulations of embryonic thermal environments, addressing widespread patterns at a comparative level has been hampered by the lack of accessible data in a format that is amendable to updates as new studies emerge. Here, we describe a database with nearly 10, 000 phenotypic estimates from 155 species of reptile, collected from 300 studies manipulating incubation temperature (published between 1974-2016). The data encompass various morphological, physiological, behavioural and performance traits along with growth rates, developmental timing, sex ratio and survival (e.g., hatching success). This resource will serve as an important data repository for addressing overarching questions about thermal plasticity of reptile embryos.

Temperature variability is predicted to increase in the coming century due to climate change. However, the biological impact of increased temperature variability on animals remains largely unexplored. Here, we experimentally exposed gravid viviparous lizards (Eremias multiocellata) to two thermal environments [constant daily maximum (CDM) versus variable daily maximum (VDM) treatment with the same average temperature] to address maternal and offspring responses to increased variability in ambient temperature. Females from the VDM treatment delayed parturition, but produced similar litter sizes and litter masses as did CDM females. Offspring from the VDM treatment selected higher body temperatures, had higher metabolic rates and higher growth rates, and grew to a similar size as those from the CDM treatment despite having a shorter growth period prior to hibernation. Therefore, phenotypic plasticity may be critical for lizards to respond effectively to climate change, and its role in responding to increasingly variable temperatures warrants further attention.

Early life environments shape phenotypic development in important ways that can lead to long-lasting effects on phenotype and fitness. In reptiles, one aspect of the early environment that impacts development is temperature (termed 'thermal developmental plasticity'). Indeed, the thermal environment during incubation is known to influence morphological, physiological, and behavioral traits, some of which have important consequences for many ecological and evolutionary processes. Despite this, few studies have attempted to synthesize and collate data from this expansive and important body of research. Here, we systematically review research into thermal developmental plasticity across reptiles, structured around the key papers and findings that have shaped the field over the past 50 years. From these papers, we introduce a large database (the 'Reptile Development Database') consisting of 9,773 trait means across 300 studies examining thermal developmental plasticity. This dataset encompasses data on a range of phenotypes, including morphological, physiological, behavioral, and performance traits along with growth rate, incubation duration, sex ratio, and survival (e.g., hatching success) across all major reptile clades. Finally, from our literature synthesis and data exploration, we identify key research themes associated with thermal developmental plasticity, important gaps in empirical research, and demonstrate how future progress can be made through targeted empirical, meta-analytic, and comparative work., (© 2018 Wiley Periodicals, Inc.)

Scientific interest in developmental plasticity spans many disciplines, and research on reptiles has provided many insights into this field. We highlight these contributions, review the field's history, and introduce the special issue on this topic ., (© 2018 Wiley Periodicals, Inc.)

The vulnerability of species to climate warming varies along latitudinal and elevational clines, but how sympatric species vary in vulnerability to climate warming remains largely unknown. We experimentally simulated nest temperatures of two sympatric lizards with divergent microhabitat preferences (Phrynocephalus przewalskii and Eremias argus), under climate warming senarios, to determine the response of embryos to increased mean temperatures and heat waves. Our study demonstrated that simulated climate warming reduced hatching success and hatchling size and growth in E. argus (that prefers closed microhabitats), but had less effect in P. przewalskii (that occupies open microhabitats). The reduced growth rate of E. argus hatchlings was associated with a decrease in metabolic rate, which was more evident in hatchling E. argus than in P. przewalskii. Our results suggest lizards that prefer closed microhabitats may be more vulnerable to climate warming than those that prefer open microhabitats; further studies are needed to test this hypothesis. More generally, the divergent responses of sympatric species to climate warming highlights the importance of distinguishing the thermal sensitivity of behavior and physiology for each species of a community, in order to make predictions about the impacts of climate warming at regional scales., (© 2018 Wiley Periodicals, Inc.)

The debate about behavioral thermoregulation inside reptile eggs centers on the frequency (and hence, biological significance) of the phenomenon, not about its validity. Both sides of the debate agree that large eggs in shallow nests laid in sun-exposed soil will experience clines in mean temperature and (especially) diel thermal variance; that embryos in the middle phase of development have the ability to reposition themselves, and room to do so; and that small changes in developmental temperatures can influence offspring fitness. Equally, all protagonists agree that thermal clines will be too low in some other kinds of nests, and that embryonic repositioning is impossible very early and very late in development. Based on an array of other fitness-enhancing behaviors exhibited by tetrapod embryos, and general principles for recognizing adaptation, we conclude that behavioral thermoregulation inside the egg likely is adaptive in some but not all reptile species. We identify productive directions for empirical research to resolve points of contention., (© 2018 Wiley Periodicals, Inc.)

Broken-symmetry density functional calculations have been performed on the [Fe a3 4+ ,Cu B 2+ ] state of the dinuclear center (DNC) for the P R → F part of the catalytic cycle of ba 3 cytochrome c oxidase (CcO) from Thermus thermophilus (Tt), using the OLYP-D3-BJ functional. The calculations show that the movement of the H 2 O molecules in the DNC affects the pK a values of the residue side chains of Tyr237 and His376 + , which are crucial for proton transfer/pumping in ba 3 CcO from Tt. The calculated lowest energy structure of the DNC in the [Fe a3 4+ ,Cu B 2+ ] state (state F) is of the form Fe a3 4+ ═O 2- ···Cu B 2+ , in which the H 2 O ligand that resulted from protonation of the OH - ligand in the P R state is dissociated from the Cu B 2+ site. The calculated Fe a3 4+ ═O 2- distance in F (1.68 Å) is 0.03 Å longer than that in P R (1.65 Å), which can explain the different Fe a3 4+ ═O 2- stretching modes in P (804 cm -1 ) and F (785 cm -1 ) identified by resonance Raman experiments. In this F state, the Cu B 2+ ···O 2- (ferryl-oxygen) distance is only around 2.4 Å. Hence, the subsequent O H state [Fe a3 3+ -OH - -Cu B 2+ ] with a μ-hydroxo bridge can be easily formed, as shown by our calculations.

Obligate brood parasites have evolved unusually thick-shelled eggs, which are hypothesized to possess a variety of functions such as resistance to puncture ejection by their hosts. In this study, we tested the hypothesis that obligate brood parasites lay unusually thick-shelled eggs to retain more heat for the developing embryo and thus contribute to early hatching of parasite eggs. By doing so, we used an infrared thermal imaging system as a non-invasive method to quantify the temperature of eggshells of common cuckoos (Cuculus canorus) and their Oriental reed warbler (Acrocephalus orientalis) hosts in an experiment that artificially altered the duration of incubation. Our results showed that cuckoo eggshells had higher temperature than host eggs during incubation, but also less fluctuations in temperature during incubation disturbance. Therefore, there was a thermal and hence a developmental advantage for brood parasitic cuckoos of laying thick-shelled eggs, providing another possible explanation for the unusually thick-shelled eggs of obligate brood parasites and earlier hatching of cuckoo eggs compared to those of the host.

The thermal environment of embryos differs significantly along a latitudinal cline, and the mechanism by which embryos respond to this geographic temperature variation has attracted increasing attention recently. Here, we carried out a common-garden experiment of egg incubation at two fluctuating temperature regimes to elucidate the latitudinal pattern and thermal dependence of the embryonic development rate and offspring performance in the Chinese soft-shelled turtle (Pelodiscus sinensis). Our results demonstrated significant temperature-by-population interactions on variations in the incubation period and hatchling righting response. The incubation period was shorter and the daily number of heart beats was higher in the medium- and high-latitude populations than in the low-latitude population at low incubation temperatures but not at high temperatures. Offspring from the medium- and high-latitude populations showed a faster righting response than those from the low-latitude population when incubated at low temperatures, whereas offspring from the high-latitude population showed a faster righting response than those from the medium-latitude population when incubated at high temperatures. This indicates that turtle embryos from different latitudinal locations may have evolved divergent thermal sensitivities of embryonic development in response to their respective nest environments.

Body size is directly linked to key life history traits such as growth, fecundity, and survivorship. Identifying the causes of body size variation is a critical task in ecological and evolutionary research. Body size variation along altitudinal gradients has received considerable attention; however, the underlying mechanisms are poorly understood. Here, we compared the growth rate and age structure of toad-headed lizards ( Phrynocephalus vlangalii ) from two populations found at different elevations in the Qinghai-Tibetan Plateau. We used mark-recapture and skeletochronological analysis to identify the potential proximate causes of altitudinal variation in body size. Lizards from the high-elevation site had higher growth rates and attained slightly larger adult body sizes than lizards from the low-elevation site. However, newborns produced by high-elevation females were smaller than those by low-elevation females. Von Bertalanffy growth estimates predicted high-elevation individuals would reach sexual maturity at an earlier age and have a lower mean age than low-elevation individuals. Relatively lower mean age for the high-elevation population was confirmed using the skeletochronological analysis. These results support the prediction that a larger adult body size of high-elevation P. vlangalii results from higher growth rates, associated with higher resource availability.

How ectotherms exploit thermal resources has important implications for their habitat utilization and thermal vulnerability to climate warming. To address this issue, we investigated thermal relations of three sympatric lizard species (Eremias argus, Eremias multiocellata, and Phrynocephalus przewalskii) in the desert steppe of Inner Mongolia, China. We determined the thermoregulatory behavior, body temperature (T b ), operative temperature (T e ), selected body temperature (T sel ), and critical thermal maximum (CT max ) of adult lizards. Based on these physiological parameters, we quantified the accuracy and effectiveness of thermoregulation as well as thermal-safety margin for these species. The three species were accurate and effective thermoregulators. The P. przewalskii preferred open habitats, and had a higher T b than the two Eremias lizards, which preferred shade habitats and shuttled more frequently between the shade and sun. This indicated that the three sympatric lizards have different thermoregulatory behavior and thermal physiology, which might facilitate their coexistence in the desert steppe ecosystem. In addition, the P. przewalskii had higher T sel and CT max , and a wider thermal-safety margin than the two Eremias lizards, suggesting that the two Eremias lizards would be more vulnerable to climate warming than P. przewalskii.

Cultivated soybeans may lose some useful genetic loci during domestication. Introgression of genes from wild soybeans could broaden the genetic background and improve soybean agronomic traits. In this study, through whole-genome sequencing of a recombinant inbred line population derived from a cross between a wild soybean ZYD7 and a cultivated soybean HN44, and mapping of quantitative trait loci for seed weight, we discovered that a phosphatase 2C-1 (PP2C-1) allele from wild soybean ZYD7 contributes to the increase in seed weight/size. PP2C-1 may achieve this function by enhancing cell size of integument and activating a subset of seed trait-related genes. We found that PP2C-1 is associated with GmBZR1, a soybean ortholog of Arabidopsis BZR1, one of key transcription factors in brassinosteroid (BR) signaling, and facilitate accumulation of dephosphorylated GmBZR1. In contrast, the PP2C-2 allele with variations of a few amino acids at the N-terminus did not exhibit this function. Moreover, we showed that GmBZR1 could promote seed weight/size in transgenic plants. Through analysis of cultivated soybean accessions, we found that 40% of the examined accessions do not have the PP2C-1 allele, suggesting that these accessions can be improved by introduction of this allele. Taken together, our study identifies an elite allele PP2C-1, which can enhance seed weight and/or size in soybean, and pinpoints that manipulation of this allele by molecular-assisted breeding may increase production in soybean and other legumes/crops., (Copyright © 2017 The Author. Published by Elsevier Inc. All rights reserved.)

Food availability significantly affects an animal's energy metabolism, and thus its phenotype, survival, and reproduction. Maternal and offspring responses to food conditions are critical for understanding population dynamics and life-history evolution of a species. In this study, we conducted food manipulation experiments in field enclosures to identify the effect of food restriction on female reproductive traits and postpartum body condition, as well as on hatchling phenotypes, in a lacertid viviparous lizard from the Inner Mongolian desert steppe of China. Females under low-food availability treatment (LFT) had poorer immune function and body condition compared with those under high-food availability treatment (HFT). The food availability treatments significantly affected the litter size and litter mass of the females, but not their gestation period in captivity or brood success, or the body size, sprint speed, and sex ratio of the neonates. Females from the LFT group had smaller litter sizes and, therefore, lower litter mass than those from the HFT group. These results suggest that female racerunners facing food restriction lay fewer offspring with unchanged body size and locomotor performance, and incur a cost in the form of poor postpartum body condition and immune function. The flexibility of maternal responses to variable food availability represents an important life strategy that could enhance the resistance of lizards to unpredictable environmental change.

Despite widespread temperature-dependent sex determination (TSD) in reptiles, it is still unclear how the molecular network responds to temperature variation and drives the sexual fate. Profiling of sex-related genes is the first step in understanding the sex determination system in reptiles. In this study, we cloned the full-length coding sequences of Cyp19a1, Foxl2, Rspo1, Sf1, and Sox9 in an Asian freshwater turtle (Mauremys reevesii) with TSD and identified the expression patterns of these genes and Dmrt1 at different incubation temperatures to understand their roles in urogenital development. Our results showed that Cyp19a1, Foxl2, and Rspo1 were expressed in the adrenal-kidney-gonadal complex at a high level in females, while Sf1 and Dmrt1 were highly expressed in males. In addition, Foxl2 and Rspo1 showed sex-dimorphic expression in the presumed early thermosensitive period (TSP), Dmrt1 was upregulated at the beginning of the presumed TSP, and Sox9 did not show sex-dimorphic expression until the end of the presumed TSP. These results suggest that Foxl2 and Rspo1 are probably upstream genes involved in female sex determination and that Dmrt1 may be a key factor in male sex determination. Therefore, our study provides a solid foundation for further investigations on the molecular mechanism underlying sex determination in M. reevesii., (© 2017 S. Karger AG, Basel.)

Light is an environmental factor that is known to profoundly affect embryonic development in some oviparous vertebrates, but such effects are unstudied in reptiles. We investigated the light sensitivity of lizard embryos by examining the thickness and light transmittance of eggshells as well as the effect of light on embryonic development and hatchling traits in four lizard species, the Chinese skink (Plestiodon chinensis), the northern grass lizard (Takydromus septentrionalis), the oriental leaf-toed gecko (Hemidactylus bowringii) and the Japanese gecko (Gekko japonicus). The eggshells were thinner and thus had higher light transmittance in Chinese skink than the other three species. Light exposure during incubation significantly accelerated the embryonic development in all species, with higher light intensity resulting in faster embryonic development. Interestingly, light stimulation negatively influenced hatchling size and survival in skinks, but had no effect in lacertids and geckos. This interspecific discrepancy not only relates to the differences in thickness and light transmittance of eggshells, but might also reflect the differences in the reproductive habits of these species. Given the diversity of light conditions that reptile embryos face during development, studies on the response of reptile embryos to light may offer a unique opportunity to understand the mechanisms of embryonic light sensitivity in animals.

Extreme high temperatures are occurring more frequently with ongoing anthropogenic climate warming, but the experimental tests of the effects of high temperatures on terrestrial vertebrates in natural conditions are rare. In this study, we investigated the effects of extreme high temperatures on female reproduction and offspring traits of multi-ocellated racerunners (Eremias multiocellata) kept in field enclosures in the desert steppe of Inner Mongolia. Our studies indicate that high temperatures significantly affect the gestation period and reproductive output of females and the offspring sex ratio, but have little impact on offspring body size and mass. More interestingly, we found that the effect of extreme high temperatures on female reproductive output was not consistent between two consecutive years that differed in precipitation. Low precipitation may aggravate the impact of climate warming on lizards and negatively affect the survival of lizards in the desert steppe. Our results provide evidence that temperature interacts with precipitation to determine the life history of lizards, and they suggest that a drier and hotter environment, such as the future climate in arid mid-latitude areas, will likely impose severe pressure on lizard populations, which are an important component of the food web in desert areas around the world.

The adaptive significance of temperature-dependent sex determination (TSD) has attracted a great deal of research, but the underlying mechanisms by which temperature determines the sex of a developing embryo remain poorly understood. Here, we manipulated the level of a thyroid hormone (TH), triiodothyronine (T 3 ), during embryonic development (by adding excess T 3 to the eggs of the red-eared slider turtle Trachemys scripta, a reptile with TSD), to test two competing hypotheses on the proximate basis for TSD: the developmental rate hypothesis versus the hormone hypothesis Exogenous TH accelerated embryonic heart rate (and hence metabolic rate), developmental rate, and rates of early post-hatching growth. More importantly, hyperthyroid conditions depressed expression of Cyp19a1 (the gene encoding for aromatase) and levels of oestradiol, and induced more male offspring. This result is contrary to the direction of sex-ratio shift predicted by the developmental rate hypothesis, but consistent with that predicted by the hormone hypothesis Our results suggest an important role for THs in regulating sex steroid hormones, and therefore, in affecting gonadal sex differentiation in TSD reptiles. Our study has implications for the conservation of TSD reptiles in the context of global change because environmental contaminants may disrupt the activity of THs, and thereby affect offspring sex in TSD reptiles., (© 2016 The Author(s).)

Cytochrome c oxidase (CcO) is a vital enzyme that catalyzes the reduction of molecular oxygen to water and pumps protons across mitochondrial and bacterial membranes. While proton uptake channels as well as water exit channels have been identified for A-type CcOs, the means by which water and protons exit B-type CcOs remain unclear. In this work, we investigate potential mechanisms for proton transport above the dinuclear center (DNC) in ba3-type CcO of Thermus thermophilus. Using long-time scale, all-atom molecular dynamics (MD) simulations for several relevant protonation states, we identify a potential mechanism for proton transport that involves propionate A of the active site heme a3 and residues Asp372, His376 and Glu126(II), with residue His376 acting as the proton-loading site. The proposed proton transport process involves a rotation of residue His376 and is in line with experimental findings. We also demonstrate how the strength of the salt bridge between residues Arg225 and Asp287 depends on the protonation state and that this salt bridge is unlikely to act as a simple electrostatic gate that prevents proton backflow. We identify two water exit pathways that connect the water pool above the DNC to the outer P-side of the membrane, which can potentially also act as proton exit transport pathways. Importantly, these water exit pathways can be blocked by narrowing the entrance channel between residues Gln151(II) and Arg449/Arg450 or by obstructing the entrance through a conformational change of residue Tyr136, respectively, both of which seem to be affected by protonation of residue His376., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Broken-symmetry density functional calculations have been performed on the [Fea3, CuB] dinuclear center (DNC) of ba3 cytochrome c oxidase from Thermus thermophilus in the states of [Fea3(3+)-(HO2)(-)-CuB(2+), Tyr237(-)] and [Fea3(4+)[double bond, length as m-dash]O(2-), OH(-)-CuB(2+), Tyr237˙], using both PW91-D3 and OLYP-D3 functionals. Tyr237 is a special tyrosine cross-linked to His233, a ligand of CuB. The calculations have shown that the DNC in these states strongly favors the protonation of His376, which is above propionate-A, but not of the carboxylate group of propionate-A. The energies of the structures obtained by constrained geometry optimizations along the O-O bond cleavage pathway between [Fea3(3+)-(O-OH)(-)-CuB(2+), Tyr237(-)] and [Fea3(4+)[double bond, length as m-dash]O(2-)HO(-)-CuB(2+), Tyr237˙] have also been calculated. The transition of [Fea3(3+)-(O-OH)(-)-CuB(2+), Tyr237(-)] → [Fea3(4+)[double bond, length as m-dash]O(2-)HO(-)-CuB(2+), Tyr237˙] shows a very small barrier, which is less than 3.0/2.0 kcal mol(-1) in PW91-D3/OLYP-D3 calculations. The protonation state of His376 does not affect this O-O cleavage barrier. The rate limiting step of the transition from state A (in which O2 binds to Fea3(2+)) to state PM ([Fea3(4+)[double bond, length as m-dash]O(2-), OH(-)-CuB(2+), Tyr237˙], where the O-O bond is cleaved) in the catalytic cycle is, therefore, the proton transfer originating from Tyr237 to O-O to form the hydroperoxo [Fea3(3+)-(O-OH)(-)-CuB(2+), Tyr237(-)] state. The importance of His376 in proton uptake and the function of propionate-A/neutral-Asp372 as a gate to prevent the proton from back-flowing to the DNC are also shown.