Novel brain gene-expression patterns are associated with a novel predaceous behaviour in tadpoles

Novel behaviours can spur evolutionary change and sometimes even precede morphological innovation, but the evolutionary and developmental contexts for their origins can be elusive. One proposed mechanism to generate behavioural innovation is a shift in the developmental timing of gene-expression patterns underlying an ancestral behaviour, or molecular heterochrony. Alternatively, novel suites of gene expression, which could provide new contexts for signalling pathways with conserved behavioural functions, could promote novel behavioural variation. To determine the relative contributions of these alternatives to behavioural innovation, I used a species of spadefoot toad, Spea bombifrons . Based on environmental cues, Spea larvae develop as either of two morphs: ‘omnivores' that, like their ancestors, feed on detritus, or ‘carnivores' that are predaceous and cannibalistic. Because all anuran larvae undergo a natural transition to obligate carnivory during metamorphosis, it has been proposed that the novel, predaceous behaviour in Spea larvae represents the accelerated activation of gene networks influencing post-metamorphic behaviours. Based on comparisons of brain transcriptional profiles, my results reject widespread heterochrony as a mechanism promoting the expression of predaceous larval behaviour. They instead suggest that the evolution of this trait relied on novel patterns of gene expression that include components of pathways with conserved behavioural functions.

Differential encoding of signals and preferences by noradrenaline in the anuran brain

ABSTRACTSocial preferences enable animals to selectively interact with some individuals over others. One influential idea for the evolution of social preferences is that preferred signals evolve because they elicit greater neural responses from sensory systems. However, in juvenile plains spadefoot toad (Spea bombifrons), a species with condition-dependent mating preferences, responses of the preoptic area, but not of the auditory midbrain, mirror adult social preferences. To examine whether this separation of signal representation from signal valuation generalizes to other anurans, we compared the relative contributions of noradrenergic signalling in the preoptic area and auditory midbrain of S. bombifrons and its close relative Spea multiplicata. We manipulated body condition in juvenile toads by controlling diet and used high pressure liquid chromatography to compare call-induced levels of noradrenaline and its metabolite MHPG in the auditory midbrain and preoptic area of the two species. We found that calls from the two species induced different levels of noradrenaline and MHPG in the auditory system, with higher levels measured in both species for the more energetic S. bombifrons call. In contrast, noradrenaline levels in the preoptic area mirrored patterns of social preferences in both S. bombifrons and S. multiplicata. That is, noradrenaline levels were higher in response to the preferred calls within each species and were modified by diet in S. bombifrons (with condition-dependent preferences) but not S. multiplicata (with condition-independent preferences). Our results are consistent with a potentially important role for preoptic noradrenaline in the development of social preferences and indicate that it could be a target of selection in the evolution of condition-dependent social preferences.

Female toads engaging in adaptive hybridization prefer high-quality heterospecifics as mates

Hybridization—interbreeding between species—is generally thought to occur randomly between members of two species. Contrary to expectation, female plains spadefoot toads (Spea bombifrons) can increase their evolutionary fitness by preferentially mating with high-quality males of another species, the Mexican spadefoot toad (Spea multiplicata). Aspects of Mexican spadefoot males’ mating calls predict their hybrid offspring’s fitness, and plains spadefoot females prefer Mexican spadefoot males on the basis of these attributes, but only in populations and ecological conditions where hybridization is adaptive. By selecting fitness-enhancing mates of another species, females increase hybridization’s benefits and exert sexual selection across species. Nonrandom mating between species can thereby increase the potential for adaptive gene flow between species so that adaptive introgression is not simply happenstance.

Competitively mediated changes in male toad calls can depend on call structure

Males of many species aggregate in large groups where they signal to attract females. These large aggregations create intense competition for mates, and the simultaneous signaling by many individuals can impair any given male’s ability to attract females. In response to this situation, male signals can be modified, either evolutionarily or facultatively, such that the detectability of the signal is enhanced. The way in which signals are modified varies among even closely related species, yet few studies have evaluated what causes such variation. Here, we address this issue using male spadefoot toads (Spea multiplicata and Spea bombifrons), which call to attract females. Using data from natural populations, we examined if, and how, male calls of 3 different call types (S. multiplicata with a slow call, S. bombifrons with a slow call, and S. bombifrons with a fast call) varied depending on competition with other males. We found that in both call types consisting of slow calls, call pulse rate decreased with increasing competition. By contrast, in the call type consisting of fast calls, call rate decreased with increasing competition. Moreover, we found that the relationship between competition and male call effort—a measure of the energy that males expend in calling––differed between the call types. Such variation in male signals in response to competition can have important implications for explaining diversity in male signals and patterns of sexual selection.

Diet alters species recognition in juvenile toads

Whether environmental effects during juvenile development can alter the ontogeny of adult mating behaviour remains largely unexplored. We evaluated the effect of diet on the early expression of conspecific recognition in spadefoot toads, Spea bombifrons . We found that juvenile toads display phonotaxis behaviour six weeks post-metamorphosis. However, preference for conspecifics versus heterospecifics emerged later and was diet dependent. Thus, the environment can affect the early development of species recognition in a way that might alter adult behaviour. Evaluating such effects is important for understanding variation in hybridization between species and the nature of species boundaries.

The role of Spadefoot Toad tadpoles in wetland trophic structure as influenced by environmental and morphological factors

Larval amphibians reach high densities in playa wetlands in the Southern Great Plains (SGP), USA, and thus may influence the entire structure and function of these ecosystems. We investigated whether both carnivorous and omnivorous morphotypes of Spadefoot Toad tadpoles (New Mexico Spadefoot, Spea multiplicata (Cope, 1863), and Plains Spadefoot, Spea bombifrons (Cope, 1863)) would exhibit a macrophagous feeding behavior that would allow them to occupy several trophic levels in playas. We also compared tadpole diets and foregut widths as influenced by the land use surrounding playas (cultivated versus grassland watersheds), year (dry versus wet year), and body size (snout-to-vent length). Tadpole diets were dominated by detritus and diatoms and tadpole foreguts increased with body size. Generally, more arthropods and less cyanobacteria were found in Spea tadpole diets as tadpoles grew larger, suggesting they influence different trophic levels with age. Foreguts were wider in carnivores than omnivores, suggesting carnivores had increased ability to ingest larger prey. Also, omnivores had wider foreguts in cropland than grassland playas, suggesting they ingest larger food items in cropland playas. From estimates of the amounts of invertebrates, detritus, and algae consumed by Spea tadpoles, we demonstrate that these larvae influence the entire trophic structure of wetland ecosystems.

Asymmetric reproductive character displacement in male aggregation behaviour

Reproductive character displacement—the evolution of traits that minimize reproductive interactions between species—can promote striking divergence in male signals or female mate preferences between populations that do and do not occur with heterospecifics. However, reproductive character displacement can affect other aspects of mating behaviour. Indeed, avoidance of heterospecific interactions might contribute to spatial (or temporal) aggregation of conspecifics. We examined this possibility in two species of hybridizing spadefoot toad (genus Spea ). We found that in Spea bombifrons sympatric males were more likely than allopatric males to associate with calling males. Moreover, contrary to allopatric males, sympatric S. bombifrons males preferentially associated with conspecific male calls. By contrast, Spea multiplicata showed no differences between sympatry and allopatry in likelihood to associate with calling males. Further, sympatric and allopatric males did not differ in preference for conspecifics. However, allopatric S. multiplicata were more variable than sympatric males in their responses. Thus, in S. multiplicata , character displacement may have refined pre-existing aggregation behaviour. Our results suggest that heterospecific interactions can foster aggregative behaviour that might ultimately contribute to clustering of conspecifics. Such clustering can generate spatial or temporal segregation of reproductive activities among species and ultimately promote reproductive isolation.