Yehonatan Alcalay

Consequences of the instar stage for behavior in a pit-building antlion

Pit-building antlion larvae are opportunistic predators that dig conical pits in loose soils, and prey on small arthropods that fall into their traps. We investigated different behavioral traits of second and third instar larvae selected for similar body masses, while also exploring the behavioral consistency and personalities of the third instar stage. Second instar larvae constructed smaller pits than third instar larvae. The former also responded more slowly to prey and exploited prey less efficiently. Notably, all these instar-based differences disappeared after molting into the third instar stage. In addition, third instar larvae exhibited consistent behavior in their pit size, response times to prey and to less extent in relocation distances. We detected two axes of behavior. The first axis included a correlation between pit size, response time and prey exploitation efficiency, thus reflecting investment in foraging activity. The second axis seemed to represent a trade-off between response time and relocation distance, implying that individuals that responded more slowly to prey, relocated over larger distances. These results point to coordinated behavior reflecting different levels of investment in foraging, while also emphasizing the importance of instar stage, in addition to body mass, when studying the behavior of such organisms characterized by a complex life cycle.

Multi-Axis Niche Examination of Ecological Specialization: Responses to Heat, Desiccation and Starvation Stress in Two Species of Pit-Building Antlions

Classical ecological studies discussing specialization usually focus on species’ performance along one niche axis. This approach may overlook niche differentiation evident in another dimension which could explain species co-occurrence. The present research exemplifies a comprehensive approach to examining local adaptation. Specifically, we examined multiple niche axes by subjecting a model organism to various experimental conditions to monitor responses to extreme stress associated with heat, desiccation and starvation. Our model system comprised two pit-building antlions: the habitat generalist Myrmeleon hyalinus and the habitat specialist Cueta lineosa. Previous research has shown that the foraging performance of the generalist is better than that of the specialist, even in the latter’s characteristic habitat. We illustrate that this apparent superiority of the habitat generalist does not manifest itself along other niche axes; rather, the habitat specialist holds a set of traits that provide an advantage under harsh environmental conditions. Specifically, C. lineosa has an advantage over M. hyalinus at high temperatures, exhibiting a higher survival rate and improved foraging success (i.e., high-temperature specialist). C. lineosa is also more efficient in its energy budget, losing less mass during starvation and gaining mass more efficiently during feeding. This superior efficiency is a result of physiological adaptations as well as behavioural responses to harsh conditions. In conclusion, our results imply that the habitat specialization of C. lineosa has not led it towards an evolutionary dead-end.

Behavioral repeatability and personality in pit-building antlion larvae under differing environmental contexts

Over the last decades, there has been growing interest among behavioral ecologists in exploring animal personalities. However, while the foraging behavior of active foragers has been extensively studied, only little is known about that of sit-and-wait predators within the personality framework. We investigated the existence of repeatability and personality in pit-building antlion larvae in the context of foraging (pit construction) and habitat selection (relocation distance and direction) over time and under three environmental contexts: thermal conditions, sand depth, and soil type. Over time, repeatability was much stronger for relocation distance than for movement directionality. Additionally, we observed positive correlations across the two levels of sand depth and soil type but not between thermal conditions. Change in substrate type may induce faster decision-making in these sand-dwelling insects or could be perceived by such insects as a more drastic alteration in their habitat. We suggest that different individuals indeed possess distinct personalities. We also suggest that repeatability should be measured at two levels: the amount of energy expenditure (distances and pit construction) and the pattern of energy expenditure (directionality). Finally, our study illustrates how differing environmental conditions can result in differing levels of plasticity, while largely preserving individual personalities.

A new mode of mitochondrial transport and polarized sorting regulated by Dynein, Milton and Miro

Intrinsic cell microtubule (MT) polarity, together with molecular motors and adaptor proteins, determines mitochondrial polarized targeting and MT-dependent transport. In polarized cells, such as neurons, mitochondrial mobility and transport require the regulation of kinesin and dynein by two adaptor proteins, Milton and Miro. Recently, we found that dynein heavy chain 64C (Dhc64C) is the primary motor protein for both anterograde and retrograde transport of mitochondria in the Drosophila bristle. In this study, we show that a molecular lesion in the Dhc64C allele that reduced bristle mitochondrial velocity generated a variant that acts as a ‘slow’ dynein in an MT-gliding assay, indicating that dynein directly regulates mitochondrial transport. We also showed that in milton-RNAi flies, mitochondrial flux into the bristle shaft, but not velocity, was significantly reduced. Surprisingly, mitochondria retrograde flux, but not net velocity, was significantly decreased in miro-RNAi flies. We thus reveal a new mode of mitochondrial sorting in polarized cell growth, whereby bi-directional mitochondrial transport undertaken exclusively by dynein is regulated by Milton in the anterograde direction and by a Miro-dependent switch to the retrograde direction. Summary: Bi-directional mitochondrial transport driven by Dynein is regulated by Milton at primary anterograde sorting, whereas Miro regulates a retrograde transport switch.

Larval stress alters dengue virus susceptibility in Aedes aegypti (L.) adult females

In addition to genetic history, environmental conditions during larval stages are critical to the development, success and phenotypic fate of the Aedes aegypti mosquito. In particular, previous studies have shown a strong genotype-by-environment component to adult mosquito body size in response to optimal vs stressed larval conditions. Here, we expand upon those results by investigating the effects of larval-stage crowding and nutritional limitation on the susceptibility of a recent field isolate of Aedes aegypti to dengue virus serotype-2. Interestingly, female mosquitoes from larvae subjected to a stressed regime exhibited significantly reduced susceptibility to disseminated dengue infection 14days post infection compared to those subjected to optimal regimes. Short term survivorship post-infected blood feeding was not significantly different. As with body size, dengue virus susceptibility of a mosquito population is determined by a combination of genetic and environmental factors and is likely maintained by balancing selection. Here, we provide evidence that under different environmental conditions, the innate immune response of field-reared mosquitoes exhibits a large range of phenotypic variability with regard to dengue virus susceptibility. Further, as with body size, our results suggest that mosquitoes reared under optimal laboratory conditions, as employed in all mosquito-pathogen studies to date, may not always be realistic proxies for natural populations.

Foraging syndromes and trait variation in antlions along a climatic gradient

Behavioral syndromes arise when individual behavior is correlated over time and/or across environmental contexts, often resulting in inter-population behavioral differences. Three main hypotheses have been suggested to explain the evolution of behavioral syndromes. The constraint hypothesis suggests that behaviors originate from a shared mechanism with a strong genetic or physiological basis. In contrast, according to the adaptive hypothesis, behavioral syndromes depend on specific selective pressures in each environment, and thus should evolve when specific behavioral combinations are advantageous. Finally, behavioral syndromes can also arise owing to neutral stochastic processes. We tested here for variation in the foraging syndromes of pit-building antlions originating from different populations along a climatic gradient. Although inter-population variation existed in some traits, foraging syndromes were similar across populations, supporting the constraint hypothesis. These findings suggest that stabilizing selection, acting on the foraging behavior of antlions during their larval phase, outweighs local selection pressures, resulting in “constraint syndromes.” We also explored behavioral repeatability of foraging-related traits within and among habitats (natural, novel and disturbed habitats), and detected different levels of repeatability: pit diameter was more repeatable than response time to prey, followed by prey exploitation efficiency. Behavioral repeatability of the same trait differed according to context, suggesting that repeatability is a trait in itself and should not be considered identical even when studying the same behavioral trait.

Slow growth improves compensation ability: examining growth rate and starvation endurance in pit-building antlions from semi-arid and hyper-arid regions

Different environments are expected to exert differential selective pressures, often generating distinct sets of traits in organisms inhabiting different geographic regions. Starvation endurance is an important trait for organisms in harsh (i.e., extreme climate and/or biotically poor) and unpredictable environments. This is especially true for sit-and-wait predators, such as antlions, which experience stronger fluctuations in prey arrivals than do actively searching predators. We conducted an experimental comparison of starvation endurance in pit-building antlions, originating from semi-arid and hyper-arid environments. We hypothesized that individuals from the climatically harsher and biotically poor environment (i.e., hyper-arid) should be better adapted to endure long starvation periods. Additionally, we posited that faster-growing individuals are expected to be more sensitive to starvation because of their need to sustain higher metabolic rates. We found that antlions originating from the semi-arid region maintained higher activity levels, which led to slightly higher mass loss rates during starvation, but enabled faster recovery when food supply was renewed. Conversely, antlions originating from the hyper-arid region had lower activity levels, consistent with their lower rate of mass loss during starvation, but this came at the expense of decreased response to prey and lower growth rate when food became available again. Each strategy holds its advantages for coping with long starvation periods, and we cannot say decisively which strategy is better. Results from both regions were consistent with the predictions of the growth compensation phenomenon: antlions that were fed less frequently pre-starvation grew at faster rates when food supply was renewed. Our study demonstrates that individuals originating from different environments adopt different strategies in order to endure starvation, exemplifying antlions’ ability to compensate for mass lost during starvation.

Dynamic microtubule organization and mitochondrial transport are regulated by distinct Kinesin-1 pathways

ABSTRACT The microtubule (MT) plus-end motor kinesin heavy chain (Khc) is well known for its role in long distance cargo transport. Recent evidence showed that Khc is also required for the organization of the cellular MT network by mediating MT sliding. We found that mutations in Khc and the gene of its adaptor protein, kinesin light chain (Klc) resulted in identical bristle morphology defects, with the upper part of the bristle being thinner and flatter than normal and failing to taper towards the bristle tip. We demonstrate that bristle mitochondria transport requires Khc but not Klc as a competing force to dynein heavy chain (Dhc). Surprisingly, we demonstrate for the first time that Dhc is the primary motor for both anterograde and retrograde fast mitochondria transport. We found that the upper part of Khc and Klc mutant bristles lacked stable MTs. When following dynamic MT polymerization via the use of GFP-tagged end-binding protein 1 (EB1), it was noted that at Khc and Klc mutant bristle tips, dynamic MTs significantly deviated from the bristle parallel growth axis, relative to wild-type bristles. We also observed that GFP-EB1 failed to concentrate as a focus at the tip of Khc and Klc mutant bristles. We propose that the failure of bristle tapering is due to defects in directing dynamic MTs at the growing tip. Thus, we reveal a new function for Khc and Klc in directing dynamic MTs during polarized cell growth. Moreover, we also demonstrate a novel mode of coordination in mitochondrial transport between Khc and Dhc. Summary: Highly polarized Drosophila bristle cells reveal that dynamic microtubule organization and mitochondrial transport are regulated by distinct Kinesin-1 pathways, and a novel mode of coordination between Khc and Dhc in mitochondrial transport.