A genetic tool kit for cellular and behavioral analyses of insect sugar receptors

Arthropods employ a large family of up to 100 putative taste or gustatory receptors (Grs) for the recognition of a wide range of non-volatile chemicals. In Drosophila melanogaster, a small subfamily of 8 Gr genes is thought to mediate the detection of sugars, the fly''s major nutritional source. However, the specific roles for most sugar Gr genes are not known. Here, we report the generation of a series of mutant sugar Gr knock-in alleles and several composite sugar Gr mutant strains, including a sugar blind strain, which will facilitate the characterization of this gene family. Using Ca²⁺ imaging experiments, we show that most gustatory receptor neurons (GRNs) of sugar blind flies (lacking all 8 sugar Gr genes) fail to respond to any sugar tested. Moreover, expression of single sugar Gr genes in most sweet GRNs of sugar-blind flies does not restore sugar responses. However, when pair-wise combinations of sugar Gr genes are introduced to sweet GRNs, responses to select sugars are restored. We also examined the cellular phenotype of flies homozygous mutant for Gr64a, a Gr gene previously reported to be a major contributor for the detection of many sugars. In contrast to these claims, we find that sweet GRNs of Gr64a homozygous mutant flies show normal responses to most sugars, and only modestly reduced responses to maltose and maltotriose. Thus, the precisely engineered genetic mutations of single Gr genes and construction of a sugar-blind strain provide powerful analytical tools for examining the roles of Drosophila and other insect sugar Gr genes in sweet taste.     

Two‐module organization of the mandible in the yellow‐necked mouse: a comparison between two different morphometric approaches

Mandibles of yellow‐necked mouse (Apodemus flavicollis) were used to explore modularity. We tested a biological hypothesis that two separate modules (alveolar region and ascending ramus) can be recognized within the mandible. As a second research goal, we compared two different morphometric procedures under the assumption that methodological approaches that use either geometric or traditional morphometric techniques should give similar results. Besides confirmation of the predicted hypothesis of modularity, the application of both approaches revealed that: (i) modularity was somewhat more evident when it was analysed on the asymmetric (fluctuating asymmetry, FA) than on the symmetric (individual variation) component of variation; (ii) there is correspondence in the patterns of individual variation and FA, which indicates that integration of mandibular traits among individuals is primarily due to direct developmental interactions; and (iii) allometry does not obscure the hypothesized modularity for individual variation or for FA. In addition, traditional morphometric method allowed us to check whether allometry influenced each module to the same extent and to conclude that the ascending ramus is more heavily influenced by general size than the alveolar region. In studies of modularity, differences in methods can lead to discrepancies in the results, and therefore, some caution is required when comparing findings from different investigations. The substantial agreement between our results provides evidence that, when considering two‐module organization of the mouse mandible, direct comparison among studies that use the methods applied herein is, in great part, reliable.     

Diet and Mandibular Morphology in African Apes

Investigations seeking to understand the relationship between mandibular form, function, and dietary behavior have focused on the mandibular corpus and symphysis. African apes vary along a gradient of folivory/frugivory, yet few studies have evaluated the morphology of the mandibular corpus and symphysis in these taxa, and the investigations have yielded mixed results. Specifically, studies using external metrics have identified differences in mandibular proportions that analysis of cortical bone distribution has not substantiated. I contribute to the ongoing debate on the relationship between jaw form and dietary behavior by comparing mandibular corporal and symphyseal shapes in African apes. Importantly, and in contrast to previous studies of African ape internal geometry, I include the Virunga mountain gorillas (Gorilla beringei beringei), the ape most specialized toward a folivorous diet. I test the hypotheses that 1) Gorilla beringei beringei always has significantly more robust mandibular corpora and symphyses, relative to mandibular length, than all other African apes and 2) all gorillas have significantly more robust mandibular corpora and symphyses, relative to mandibular length, than Pan. Results demonstrate that the folivorous mountain gorillas consistently exhibit a relatively wider mandibular symphysis and corpus than all other African apes. Furthermore, all gorillas consistently exhibit relatively more robust mandibular corporal and symphyseal dimensions than Pan. The results indicate that among African apes, mountain gorillas are better able to counter lateral transverse bending (wishboning) loads at the symphysis and torsional loads at the corpus. All gorillas are likewise better able to resist wishboning and vertical bending at the symphysis, and sagittal bending and torsion at the corpus, than Pan, findings that are consistent with masticating relatively tougher foods, repetitive loading of the jaws, or both. I offer possible explanations for the lack of concordance in results between studies that have analyzed the biomechanical properties of African ape mandibles and others that have relied on external metrics. More comprehensive study of the internal geometry of the mandible is needed to resolve whether African apes differ morphologically in ways predicted by dietary variation.

Morphological and biomechanical disparity of crocodile-line archosaurs following the end-Triassic extinction

Mesozoic crurotarsans exhibited diverse morphologies and feeding modes, representing considerable ecological diversity, yet macroevolutionary patterns remain unexplored. Here, we use a unique combination of morphological and biomechanical disparity metrics to quantify the ecological diversity and trophic radiations of Mesozoic crurotarsans, using the mandible as a morpho-functional proxy. We recover three major trends. First, the diverse assemblage of Late Triassic crurotarsans was morphologically and biomechanically disparate, implying high levels of ecological variation; but, following the end-Triassic extinction, disparity declined. Second, the Jurassic radiation of marine thalattosuchians resulted in very low morphological disparity but moderate variation in jaw biomechanics, highlighting a hydrodynamic constraint on mandibular form. Third, during the Cretaceous terrestrial radiations of neosuchians and notosuchians, mandibular morphological variation increased considerably. By the Late Cretaceous, crocodylomorphs evolved a range of morphologies equalling Late Triassic crurotarsans. By contrast, biomechanical disparity in the Cretaceous did not increase, essentially decoupling from morphology. This enigmatic result could be attributed to biomechanical evolution in other anatomical regions (e.g. cranium, dentition or postcranium), possibly releasing the mandible from selective pressures. Overall, our analyses reveal a complex relationship between morphological and biomechanical disparity in Mesozoic crurotarsans that culminated in specialized feeding ecologies and associated lifestyles.     

Correlation between the size,shape and position of the teeth on the jaws and the bite force in Theropoda

Theropods have fascinated both paleontologists and the general public due to their large diversity of sizes and morphologies. They also present a large variation in tooth morphologies. Previous studies have estimated the bite force of several specimens. The goal of this study is to determine if there is a correlation between the tooth size, shape and position on the skull and mandible and the bite force of these dinosaurs. Measurements were made on several theropods, including the bending strength of the teeth on the anterior-posterior and the mediolateral axes of the jaws, as well as the bending strength of the mandible, and were compared to fossil and modern Crocodylia. We observed that several bending strength maxima of the teeth trends were aligned with key areas of the mandible, and that the size, shape as well as the position of the teeth on the jaws were correlated with the bite force of both Crocodylia and theropods, which can be related to their diet and feeding habits.     

Quantitative Comparison of Ontogenetic and Phylogenetic Character Changes in the Synapsid Mandible and Auditory Region

The dual transformations of the post-dentary bones into sound conducting elements of the basicranium in synapsid evolutionary and developmental histories are quantitatively compared using a modified version of a previously published methodology. Sensitivity analyses were performed to test the effects of using different phylogenies and statistics. The relative sequences of discrete osteological character transformations of the mandible and auditory region compared between the ontogeny of Monodelphis domestica. the gray short-tailed opossum, and two phylogenetic hypotheses for synapsids are statistically correlated using two different tests (Spearman's R correlation and Kendall's Tau). Presumably, the characters examined in this study form a functionally integrated morphological unit that is distinct throughout evolutionary and developmental history. Assuming a von Baerian developmental pattern, the methodology discussed here could potentially be used as a test of similarity between phylogenetic hypotheses; however, further work needs to be done to test this potential.     

A statistical framework for testing modularity in multidimensional data

Modular variation of multivariate traits results from modular distribution of effects of genetic and epigenetic interactions among those traits. However, statistical methods rarely detect truly modular patterns, possibly because the processes that generate intramodular associations may overlap spatially. Methodologically, this overlap may cause multiple patterns of modularity to be equally consistent with observed covariances. To deal with this indeterminacy, the present study outlines a framework for testing a priori hypotheses of modularity in which putative modules are mathematically represented as multidimensional subspaces embedded in the data. Model expectations are computed by subdividing the data into arrays of variables, and intermodular interactions are represented by overlapping arrays. Covariance structures are thus modeled as the outcome of complex and nonorthogonal intermodular interactions. This approach is demonstrated by analyzing mandibular modularity in nine rodent species. A total of 620 models are fit to each species, and the most strongly supported are heuristically modified to improve their fit. Five modules common to all species are identified, which approximately map to the developmental modules of the mandible. Within species, these modules are embedded within larger "super-modules," suggesting that these conserved modules act as building blocks from which covariation patterns are built.     

Influences of geometrical and mechanical properties of bone tissues in mandible behaviour – experimental and numerical predictions

The properties and geometry of bone in the mandible play a key role in mandible behaviour during a person’s lifetime, and attention needs to be paid to the influence of bone properties. We analysed the effect of bone geometry, size and bone properties in mandible behaviour, experimenting on cadaveric mandibles and FE models. The study was developed using the geometry of a cadaveric mandible without teeth. Three models of cadaveric condyles were experimentally tested with instrumented with four rosettes, and a condyle reaction of 300 N. Four finite element models were considered to validate the experiments and analyse mandible behaviour. One numeric model was simulated with 10 muscles in a quasi-static condition. The experimental results present different condyle stiffness’s, of 448, 215 and 254 N/mm. The values presented in the rosettes are influenced by bone geometry and bone thickness; maximum value was ?600 με in rosette #4, and the maximum strain difference between mandibles was 111%. The numerical results show that bone density decreases and strain distribution increases in the thinner mandible regions. Nevertheless, the global behaviour of the structure remains similar, but presents different strain magnitudes. The study shows the need to take into account bone characteristics and their evolutions in order to improve implant design and fixation throughout the patient life. The change in bone stiffness promotes a change in maximum strain distribution with same global behaviour.     

Feeding behaviour,midgut distension and ovarian development in Phlebotomus papatasi (Diptera: Psychodidae)

Phlebotomus papatasi females were fed through membranes or from cotton wool soaked in blood, water, sucrose or sodium chloride solutions. In membrane-fed flies, all diets were routed to the midgut and not to the crop. Following the meals that went to the midgut, females showed ovarian development at least 3 times greater than in sucrose-fed, autogenous control flies. Neither small quantities of water arriving in the midgut following drinking from soaked cotton wool, nor piercing of a membrane without feeding, stimulated ovarian development. Flies exhibited different feeding behaviour namely, blood feeding, sugar feeding, and water drinking. The blood-feeding behaviour was typical of flies ingesting any of the experimental diets through membranes, or blood or saline from cotton wool. The other two types of behaviour were observed in flies which fed from soaked cotton wool. The type of behaviour was characterized by the depth of penetration of the mouthparts into the substrate, the deployment of the palps and the degree of contact between the palps and the surface. It is suggested that the stimuli which control the routing of meals to the crop or to the midgut are derived from these types of behaviour.     

Territorial aggressiveness and predation: two possible origins of snapping in the ant Plectroctena minor

Plectroctena minor workers have long mandibles that can snap and deliver a sharp blow to intruders or prey, stunning or killing them. Encounters between homocolonial P. minor workers separated for 24 h or 15 days never resulted in snapping, while this behaviour was always noted during encounters between heterocolonial workers on neutral arenas or on the territory of a colony. In the latter case, only the aliens, that generally tried to escape, were snapped at. Snapping also occurred during encounters with workers belonging to sympatric ponerine species. During predation, the percentages of snapping varied according to prey nature, suggesting prey discrimination. Termite soldiers were always snapped at, while other prey were more often snapped close to rather than far from the nest entrances, indicating an intermingling of territorial aggressiveness and predatory behaviour. We discuss the adaptive value of snapping for hunting in galleries.