Systemic orchestration of cell size throughout the body: influence of sex and rapamycin exposure in Drosophila melanogaster.

Along with differences in life histories, metazoans have also evolved vast differences in cellularity, involving changes in the molecular pathways controlling the cell cycle. The extent to which the signalling network systemically determines cellular composition throughout the body and whether tissue cellularity is organized locally to match tissue-specific functions are unclear. We cultured genetic lines of Drosophila melanogaster on food with and without rapamycin to manipulate the activity of target of rapamycin (TOR)/insulin pathways and evaluate cell-size changes in five types of adult cells: wing and leg epidermal cells, ommatidial cells, indirect flight muscle cells and Malpighian tubule epithelial cells. Rapamycin blocks TOR multiprotein complex 1, reducing cell growth, but this effect has been studied in single cell types. As adults, rapamycin-treated flies had smaller bodies and consistently smaller cells in all tissues. Regardless, females eclosed with larger bodies and larger cells in all tissues than males. Thus, differences in TOR activity and sex were associated with the orchestration of cell size throughout the body, leading to differences in body size. We postulate that the activity of TOR/insulin pathways and their effects on cellularity should be considered when investigating the origin of ecological and evolutionary patterns in life histories.

Thermal and oxygen conditions during development cause common rough woodlice (Porcellio scaber) to alter the size of their gas-exchange organs.

Terrestrial isopods have evolved pleopodal lungs that provide access to the rich aerial supply of oxygen. However, isopods occupy conditions with wide and unpredictable thermal and oxygen gradients, suggesting that they might have evolved adaptive developmental plasticity in their respiratory organs to help meet metabolic demand over a wide range of oxygen conditions. To explore this plasticity, we conducted an experiment in which we reared common rough woodlice (Porcellio scaber) from eggs to maturation at different temperatures (15 and 22 °C) combined with different oxygen levels (10% and 22% O2). We sampled animals during development (only females) and then examined mature adults (both sexes). We compared woodlice between treatments with respect to the area of their pleopod exopodites (our proxy of lung size) and the shape of Bertalanffy's equations (our proxy of individual growth curves). Generally, males exhibited larger lungs than females relative to body size. Woodlice also grew relatively fast but achieved a decreased asymptotic body mass in response to warm conditions; the oxygen did not affect growth. Under hypoxia, growing females developed larger lungs compared to under normoxia, but only in the late stage of development. Among mature animals, this effect was present only in males. Woodlice reared under warm conditions had relatively small lungs, in both developing females (the effect was increased in relatively large females) and among mature males and females. Our results demonstrated that woodlice exhibit phenotypic plasticity in their lung size. We suggest that this plasticity helps woodlice equilibrate their gas exchange capacity to differences in the oxygen supply and metabolic demand along environmental temperature and oxygen gradients. The complex pattern of plasticity might indicate the effects of a balance between water conservation and oxygen uptake, which would be especially pronounced in mature females that need to generate an aqueous environment inside their brood pouch.

An evolutionary solution of terrestrial isopods to cope with low atmospheric oxygen levels.

The evolution of current terrestrial life was founded by major waves of land invasion coinciding with high atmospheric oxygen content. These waves were followed by periods with substantially reduced oxygen concentration and accompanied by the evolution of novel traits. Reproduction and development are limiting factors for evolutionary water-land transitions, and brood care has probably facilitated land invasion. Peracarid crustaceans provide parental care for their offspring by brooding the early stages within the motherly brood pouch, the marsupium. Terrestrial isopod progeny begin ontogenetic development within the marsupium in water, but conclude development within the marsupium in air. Our results for progeny growth until hatching from the marsupium provide evidence for the limiting effects of oxygen concentration and for a potentially adaptive solution. Inclusion of air within the marsupium compensates for initially constrained growth in water through catch-up growth, and it may explain how terrestrial isopods adapted to short- and long-term changes in oxygen concentration.

Mass scaling of metabolic rates in carabid beetles (Carabidae) - the importance of phylogeny, regression models and gas exchange patterns.

The origin of the allometric relationship between standard metabolic rate (MR) and body mass (M), often described as MR=aMb , remains puzzling, and interpretation of the mass-scaling exponent, b may depend on the methodological approach, shapes of residuals, coefficient of determination (r2) and sample size. We investigated the mass scaling of MRs within and between species of Carabidae beetles. We used ordinary least squares (OLS) regression, phylogenetically generalized least squares (PGLS) regression and standardized major axis (SMA) regression to explore the effects of different model-fitting methods and data clustering caused by phylogenetic clades (grade shift) and gas exchange patterns (discontinuous, cyclic and continuous). At the interspecific level, the relationship between MR and M was either negatively allometric (b<1) or isometric (b=1), depending on the fitting method. At the intraspecific level, the relationship either did not exist or was isometric or positively allometric (b>1), and the fit was significantly improved after the analyzed dataset was split according to gas exchange patterns. The studied species originated from two distinct phylogenetic clades that had different intercepts but a common scaling exponent (OLS, 0.61) that was much shallower than the scaling exponent for the combined dataset for all species (OLS, 0.71). The best scaling exponent estimates were obtained by applying OLS while accounting for grade shifts or by applying PGLS. Overall, we show that allometry of MR in insects can depend heavily on the model fitting method, the structure of phylogenetic non-independence and ecological factors that elicit different modes of gas exchange.

The thermal environment of the nest affects body and cell size in the solitary red mason bee (Osmia bicornis L.).

Many ectotherms grow larger at lower temperatures than at higher temperatures. This pattern, known as the temperature-size rule, is often accompanied by plastic changes in cell size, which can mechanistically explain the thermal dependence of body size. However, the theory predicts that thermal plasticity in cell size has adaptive value for ectotherms because there are different optimal cell-membrane-to-cell-volume ratios at different temperatures. At high temperatures, the demand for oxygen is high; therefore, a large membrane surface of small cells is beneficial because it allows high rates of oxygen transport into the cell. The metabolic costs of maintaining membranes become more important at low temperatures than at high temperatures, which favours large cells. In a field experiment, we manipulated the thermal conditions inside nests of the red mason bee, a solitary bee that does not regulate the temperature in its nests and whose larvae develop under ambient conditions. We assessed the effect of temperature on body mass and ommatidia size (our proxy of cell size). The body and cell sizes decreased in response to a higher mean temperature and greater temperature fluctuations. This finding is in accordance with predictions of the temperature-size rule and optimal cell size theory and suggests that both the mean temperature and the magnitude of temperature fluctuations are important for determining body and cell sizes. Additionally, we observed that males of the red mason bee tend to have larger ommatidia in relation to their body mass than females, which might play an important role during mating flight.

Genetic components in a thermal developmental plasticity of the beetle Tribolium castaneum.

Low developmental temperatures cause ectotherms to retard growth, postpone maturation, and emerge at either larger or smaller adult size. In this study, we explored how these thermal responses evolved, focusing on their genetic basis. We applied a full diallel breeding design on inbred lines of the flour beetle, Tribolium castaneum. To assess the proportional contributions of genetic and non-genetic effects, each genotype, a unique combination of parental haplotypes, was reared from an egg to imago at five developmental temperatures. Faster development of females vs. males was associated with comparatively larger body masses of females (pupae and imago). In contrast, the rapid development caused by warmer environments resulted in smaller beetles (pupae and imago), but there were significant differences in this trait among genotypes. Independent effects of parental haplotypes played the major role in explaining the variance of body mass, but interactive effects of parental haplotypes explained most of the variance in developmental length. Genotypes responded to the thermal environment in a markedly uniform way. Nevertheless, we found the low statistically significant variance in the slopes of thermal reaction norms for body mass and developmental, which was mainly driven by the interactive effects of parental haplotypes. Overall, the thermal plasticity of T. castaneum follows the most common pattern among ectotherms, the so-called temperature-size rule. Detection of the low genetic variance in the shape of this response supports the idea that thermal developmental plasticity remains under a strong selective pressure in ectotherms.

Physical mechanism or evolutionary trade-off? Factors dictating the relationship between metabolic rate and ambient temperature in carabid beetles.

The tight association between ambient temperature (T) and metabolic rate (MR) is a common occurrence in ectotherms, but the determinants of this association are not fully understood. This study examined whether the relationship between MR and T is the same among individuals, as predicted by the Universal Temperature Dependence hypothesis, or whether this relationship differs between them. We used flow-through respirometry to measure standard MR and to determine gas exchange patterns for 111 individuals of three Carabidae species which differ in size (Abax ovalis, Carabus linnei and C. coriaceus), exposed to four different temperatures (ten individuals of each species measured at 6, 11, 16 and 21°C). We found a significant interaction between ln body mass and the inverse of temperature, indicating that in a given species, the effect of temperature on MR was weaker in larger individuals than in smaller individuals. Overall, this finding shows that the thermal dependence of MR is not body mass invariant. We observed three types of gas exchange patterns among beetles: discontinuous, cyclic and continuous. Additionally, the appearance of these patterns was associated with MR and T. Evolution in diverse terrestrial environments could affect diverse ventilation patterns, which accommodate changes in metabolism in response to temperature variation. In conclusion, explaining the variance in metabolism only through fundamental physical laws of thermodynamics, as predicted by the Universal Temperature Dependence hypothesis, appears to oversimplify the complexity of nature, ignoring evolutionary trade-offs that should be taken into account in the temperature - metabolism relationship.

Change in abundance of three phytophagous mite species (Acari: Eriophyidae, Tetranychidae) on quackgrass in the presence of choke disease.

Phytophagous mites and endophytic fungi may interact when sharing a host plant, potentially influencing one another's growth or population dynamics; however, interactions between them are poorly known and remain largely unexplored. In this study, quantitative associations between three species of phytophagous mites and the endophytic fungus Epichloë bromicola Leuchtm. & Schardl (Clavicipitaceae, Ascomycotina) on quackgrass, Elymus repens (L.) Gould are reported. The mites' abundance was assessed on field-collected grass shoots that were either exhibiting choke disease symptoms or without the fungus. Overall, the abundance of Tetranychus urticae and Aculodes mckenziei was significantly lower on quackgrass plants infected by E. bromicola compared to plants without the fungus. Conversely, populations of Abacarus hystrix were significantly larger on plants colonised by the fungus than on uninfected plants. Thus, the presence of this endophytic fungus may have divergent effects on different phytophagous mite species although the basis of these effects is not yet known.

Flies developed smaller cells when temperature fluctuated more frequently.

Changes in cell size might be an important component of adaptation to thermal heterogeneity. Although Drosophila melanogaster develops smaller cells at fluctuating temperatures, we do not know whether this response depends on the frequency or amplitude of thermal change. In a laboratory experiment, we exposed flies to either frequent or infrequent fluctuations between 17 and 27 °C, while controlling the total exposure to each temperature. Flies emerged from these treatments with similar body sizes, but flies at more frequent fluctuations emerged earlier and had smaller epidermal cells for a given body size. Tissue built from small cells has more nuclei for transcription, shorter distances between cell compartments, and a larger surface area for transport across membranes. Therefore, we hypothesize that physiological effects of small cells reduce lags in metabolic activity and enhance performance of flies during warming. For plasticity of cell size to confer a fitness advantage, this hypothetical benefit must outweigh the cost of maintaining a greater area of plasma membrane.

The Temperature-Size Rule in Lecane inermis (Rotifera) is adaptive and driven by nuclei size adjustment to temperature and oxygen combinations.

The evolutionary implications of the Temperature-Size Rule (TSR) in ectotherms is debatable; it is uncertain whether size decrease with temperature increase is an adaptation or a non-adaptive by-product of some temperature-dependent processes. We tested whether (i) the size of the rotifer Lecane inermis affects fecundity in a way that depends on the combination of low or high temperature and oxygen content and (ii) the proximate mechanism underlying TSR in this species is associated with nuclei size adjustment (a proxy of cell size). Small-type and large-type rotifers were obtained by culturing at different temperatures prior to the experiment and then exposed to combinations of two temperature and two oxygen conditions. Fecundity was estimated and used as a measure of fitness. Nuclei and body sizes were measured to examine the response to both environmental factors tested. The results show the following for L. inermis. (i) Body size affects fecundity in response to both temperature and oxygen, supporting a hypothesis regarding the contribution of oxygen in TSR. (ii) Large individuals are generally more fecund than small ones; however, under a combination of high temperature and poor oxygen conditions, small individuals are more fecund than large ones, in accordance with a hypothesis of the adaptive significance of TSR. (iii) The body size response to temperature is realised by nuclei size adjustment. (iv) Nuclei size changes in response to temperature and oxygen conditions, in agreement with hypotheses on the cellular mechanism underlying TSR and on a contribution of oxygen availability in TSR. These results serve as empirical evidence for the adaptive significance of TSR and validation of the cellular mechanism for the observed response.

Heat tolerance in Drosophila melanogaster is influenced by oxygen conditions and mutations in cell size control pathways.

Understanding metabolic performance limitations is key to explaining the past, present and future of life. We investigated whether heat tolerance in actively flying Drosophila melanogaster is modified by individual differences in cell size and the amount of oxygen in the environment. We used two mutants with loss-of-function mutations in cell size control associated with the target of rapamycin (TOR)/insulin pathways, showing reduced (mutant rictorΔ2) or increased (mutant Mnt1) cell size in different body tissues compared to controls. Flies were exposed to a steady increase in temperature under normoxia and hypoxia until they collapsed. The upper critical temperature decreased in response to each mutation type as well as under hypoxia. Females, which have larger cells than males, had lower heat tolerance than males. Altogether, mutations in cell cycle control pathways, differences in cell size and differences in oxygen availability affected heat tolerance, but existing theories on the roles of cell size and tissue oxygenation in metabolic performance can only partially explain our results. A better understanding of how the cellular composition of the body affects metabolism may depend on the development of research models that help separate various interfering physiological parameters from the exclusive influence of cell size. This article is part of the theme issue 'The evolutionary significance of variation in metabolic rates'.

Evolution and development of Drosophila melanogaster under different thermal conditions affected cell sizes and sensitivity to paralyzing hypoxia.

Environmental gradients cause evolutionary and developmental changes in the cellular composition of organisms, but the physiological consequences of these effects are not well understood. Here, we studied experimental populations of Drosophila melanogaster that had evolved in one of three selective regimes: constant 16 °C, constant 25 °C, or intergenerational shifts between 16 °C and 25 °C. Genotypes from each population were reared at three developmental temperatures (16 °C, 20.5 °C, and 25 °C). As adults, we measured thorax length and cell sizes in the Malpighian tubules and wing epithelia of flies from each combination of evolutionary and developmental temperatures. We also exposed flies from these treatments to a short period of nearly complete oxygen deprivation to measure hypoxia tolerance. For genotypes from any selective regime, development at a higher temperature resulted in smaller flies with smaller cells, regardless of the tissue. At every developmental temperature, genotypes from the warm selective regime had smaller bodies and smaller wing cells but had larger tubule cells than did genotypes from the cold selective regime. Genotypes from the fluctuating selective regime were similar in size to those from the cold selective regime, but their cells of either tissue were the smallest among the three regimes. Evolutionary and developmental treatments interactively affected a fly's sensitivity to short-term paralyzing hypoxia. Genotypes from the cold selective regime were less sensitive to hypoxia after developing at a higher temperature. Genotypes from the other selective regimes were more sensitive to hypoxia after developing at a higher temperature. Our results show that thermal conditions can trigger evolutionary and developmental shifts in cell size, coupled with changes in body size and hypoxia tolerance. These patterns suggest links between the cellular composition of the body, levels of hypoxia within cells, and the energetic cost of tissue maintenance. However, the patterns can be only partially explained by existing theories about the role of cell size in tissue oxygenation and metabolic performance.

Flies developed small bodies and small cells in warm and in thermally fluctuating environments.

Although plasma membranes benefit cells by regulating the flux of materials to and from the environment, these membranes cost energy to maintain. Because smaller cells provide relatively more membrane area for transport, ectotherms that develop in warm environments should consist of small cells despite the energetic cost. Effects of constant temperatures on cell size qualitatively match this prediction, but effects of thermal fluctuations on cell size are unknown. Thermal fluctuations could favour either small or large cells; small cells facilitate transport during peaks in metabolic demand whereas large cells minimize the resources needed for homeoviscous adaptation. To explore this problem, we examined effects of thermal fluctuations during development on the size of epidermal cells in the wings of Drosophila melanogaster. Flies derived from a temperate population were raised at two mean temperatures (18 and 25°C), with either no variation or a daily variation of ±4°C. Flies developed faster at a mean temperature of 25°C. Thermal fluctuations sped development, but only at 18°C. An increase in the mean and variance of temperature caused flies to develop smaller cells and wings. Thermal fluctuations reduced the size of males at 18°C and the size of females at 25°C. The thorax, the wings and the cells decreased with an increase in the mean and in the variance of temperature, but the response of cells was the strongest. Based on this pattern, we hypothesize that development of the greater area of membranes under thermal fluctuations provides a metabolic advantage that outweighs the greater energetic cost of remodelling membranes.

Rapamycin supplementation of Drosophila melanogaster larvae results in less viable adults with smaller cells.

The intrinsic sources of mortality relate to the ability to meet the metabolic demands of tissue maintenance and repair, ultimately shaping ageing patterns. Anti-ageing mechanisms compete for resources with other functions, including those involved in maintaining functional plasma membranes. Consequently, organisms with smaller cells and more plasma membranes should devote more resources to membrane maintenance, leading to accelerated intrinsic mortality and ageing. To investigate this unexplored trade-off, we reared Drosophila melanogaster larvae on food with or without rapamycin (a TOR pathway inhibitor) to produce small- and large-celled adult flies, respectively, and measured their mortality rates. Males showed higher mortality than females. As expected, small-celled flies (rapamycin) showed higher mortality than their large-celled counterparts (control), but only in early adulthood. Contrary to predictions, the median lifespan was similar between the groups. Rapamycin administered to adults prolongs life; thus, the known direct physiological effects of rapamycin cannot explain our results. Instead, we invoke indirect effects of rapamycin, manifested as reduced cell size, as a driver of increased early mortality. We conclude that cell size differences between organisms and the associated burdens of plasma membrane maintenance costs may be important but overlooked factors influencing mortality patterns in nature.

Systemic changes in cell size throughout the body of Drosophila melanogaster associated with mutations in molecular cell cycle regulators.

Along with different life strategies, organisms have evolved dramatic cellular composition differences. Understanding the molecular basis and fitness effects of these differences is key to elucidating the fundamental characteristics of life. TOR/insulin pathways are key regulators of cell size, but whether their activity determines cell size in a systemic or tissue-specific manner awaits exploration. To that end, we measured cells in four tissues in genetically modified Drosophila melanogaster (rictorΔ2 and Mnt1) and corresponding controls. While rictorΔ2 flies lacked the Rictor protein in TOR complex 2, downregulating the functions of this element in TOR/insulin pathways, Mnt1 flies lacked the transcriptional regulator protein Mnt, weakening the suppression of downstream signalling from TOR/insulin pathways. rictorΔ2 flies had smaller epidermal (leg and wing) and ommatidial cells and Mnt1 flies had larger cells in these tissues than the controls. Females had consistently larger cells than males in the three tissue types. In contrast, dorsal longitudinal flight muscle cells (measured only in males) were not altered by mutations. We suggest that mutations in cell cycle control pathways drive the evolution of systemic changes in cell size throughout the body, but additional mechanisms shape the cellular composition of some tissues independent of these mutations.

Oxygen and temperature affect cell sizes differently among tissues and between sexes of Drosophila melanogaster.

Spatio-temporal gradients in thermal and oxygen conditions trigger evolutionary and developmental responses in ectotherms' body size and cell size, which are commonly interpreted as adaptive. However, the evidence for cell-size responses is fragmentary, as cell size is typically assessed in single tissues. In a laboratory experiment, we raised genotypes of Drosophila melanogaster at all combinations of two temperatures (16 °C or 25 °C) and two oxygen levels (10% or 22%) and measured body size and the sizes of cells in different tissues. For each sex, we measured epidermal cells in a wing and a leg and ommatidial cells of an eye. For males, we also measured epithelial cells of a Malpighian tubule and muscle cells of a flight muscle. On average, females emerged at a larger body size than did males, having larger cells in all tissues. Flies of either sex emerged at a smaller body size when raised under warm or hypoxic conditions. Development at 25 °C resulted in smaller cells in most tissues. Development under hypoxia resulted in smaller cells in some tissues, especially among females. Altogether, our results show thermal and oxygen conditions trigger shifts in adult size, coupled with the systemic orchestration of cell sizes throughout the body of a fly. The nature of these patterns supports a model in which an ectotherm adjusts its life-history traits and cellular composition to prevent severe hypoxia at the cellular level. However, our results revealed some inconsistencies linked to sex, cell type, and environmental parameters, which suggest caution in translating information obtained for single type of cells to the organism as a whole.

Heat stress during development makes antlion larvae more responsive to vibrational cues.

We investigated the effects of heat stress on the responsiveness to vibrational cues, our measure of perceptual ability, in Myrmeleon bore antlion larvae (Neuroptera: Myrmeleontidae). We reared these trap-building predatory larvae under 2 heat stress regimes (mild, 30°C, and harsh, 36°C), and after they progressed from one instar stage to another, we tested their perceptual ability in common unchallenging conditions. We hypothesized that exposure to the harsh heat stress regime would impose costs resulting in handicapped vibration responsiveness. We found that the harsh heat stress regime generated more stressful conditions for the larvae, as evidenced by increased mortality and postponed molting, and the loss of body mass among larger larvae. Furthermore, among the individuals who remained alive, those originating from the harsh heat stress regime were characterized by higher vibration responsiveness. Our results suggest 2 not mutually exclusive scenarios. Costly heat stress conditions can sieve out individuals characterized by poor perceptual ability or surviving individuals can attempt to hunt more efficiently to compensate for the physiological imbalance caused by heat stress. Both of these mechanisms fit into the ongoing debate over how adaptation and plasticity contribute to shaping insect communities exposed to heat stress.

Thermal and Oxygen Flight Sensitivity in Ageing Drosophila melanogaster Flies: Links to Rapamycin-Induced Cell Size Changes.

Ectotherms can become physiologically challenged when performing oxygen-demanding activities (e.g., flight) across differing environmental conditions, specifically temperature and oxygen levels. Achieving a balance between oxygen supply and demand can also depend on the cellular composition of organs, which either evolves or changes plastically in nature; however, this hypothesis has rarely been examined, especially in tracheated flying insects. The relatively large cell membrane area of small cells should increase the rates of oxygen and nutrient fluxes in cells; however, it does also increase the costs of cell membrane maintenance. To address the effects of cell size on flying insects, we measured the wing-beat frequency in two cell-size phenotypes of Drosophila melanogaster when flies were exposed to two temperatures (warm/hot) combined with two oxygen conditions (normoxia/hypoxia). The cell-size phenotypes were induced by rearing 15 isolines on either standard food (large cells) or rapamycin-enriched food (small cells). Rapamycin supplementation (downregulation of TOR activity) produced smaller flies with smaller wing epidermal cells. Flies generally flapped their wings at a slower rate in cooler (warm treatment) and less-oxygenated (hypoxia) conditions, but the small-cell-phenotype flies were less prone to oxygen limitation than the large-cell-phenotype flies and did not respond to the different oxygen conditions under the warm treatment. We suggest that ectotherms with small-cell life strategies can maintain physiologically demanding activities (e.g., flight) when challenged by oxygen-poor conditions, but this advantage may depend on the correspondence among body temperatures, acclimation temperatures and physiological thermal limits.

Thermal Preferences of Cowpea Seed Beetles (Callosobruchus maculatus): Effects of Sex and Nuptial Gift Transfers.

The thermal environment influences insect performance, but the factors affecting insect thermal preferences are rarely studied. We studied Callosobruchus maculatus seed beetles and hypothesized that thermal preferences are influenced by water balance, with individuals with limited water reserves preferring cooler habitats to reduce evaporative water loss. Adult C. maculatus, in their flightless morph, do not consume food or water, but a copulating male provides a female with a nuptial gift of ejaculate containing nutrients and water. We hypothesized that gift recipients would prefer warmer habitats than gift donors and that both sexes would plastically adjust their thermal preferences according to the size of the transferred gift. We measured the thermal preference in each sex in individuals that were mated once or were unmated. In the mated group, we measured the sizes of the nuptial gifts and calculated proportional body mass changes in each mate during copulation. Supporting the role of water balance in thermal preference, females preferred warmer habitats than males. Nevertheless, thermal preferences in either sex were not affected by mating status or gift size. It is likely that high rates of mating and gift transfers in C. maculatus living under natural conditions promoted the evolution of constitutive sex-dependent thermal preferences.

Past thermal conditions affect hunting behaviour in larval antlions.

Some sit-and-wait predators, such as antlion larvae, construct traps to capture passing prey. The location of these traps depends on many abiotic and biotic factors, including temperature and the presence of conspecifics, which probably stimulate behaviours that minimize the costs and maximize the benefits of trap building. Here, we exposed second instar antlion larvae to elevated temperatures of 25°C (mild treatment) or 31°C (harsh treatment) for one month and then transferred them to common conditions (20°C) to examine the effects of previous thermal treatment on aggregation tendency and trap size. We predicted that antlions that experienced harsh conditions would subsequently increase the neighbouring distance and trap diameter to reduce competition with conspecifics and improve prey capture success, compensating for past conditions. In contrast with these predictions, antlions exposed to harsh conditions displayed a trend in the opposite direction, towards the decreased neighbouring distance. Furthermore, some of these antlions also built smaller traps. We discuss possible reasons for our results. The effects of previous thermal exposure have rarely been considered in terms of trap construction in antlions. Described effects may possibly apply to other sit-and-wait predators and are significant considering that many of these predators are long-lived.