The flying insect thoracic cuticle is heterogenous in structure and in thickness-dependent modulus gradation

Abstract

The thorax is a specialized structure central to an insects ability to fly. In the thorax, flight muscles are surrounded by a thin layer of cuticle. The structure, composition, and material properties of this chitinous structure may influence the efficiency of the thorax in flight. However, these properties, as well as their variation throughout anatomical regions of the thorax or between insect taxa, are not known. In this work, we provide a multi-faceted assessment of thorax cuticle for fliers with asynchronous (honey bee; Apis mellifera) and synchronous (hawkmoth; Manduca sexta) muscles. We investigated cuticle structure using histology, material composition through confocal laser scanning microscopy, and modulus gradation with nanoindentation. Our results suggest that cuticle properties of the thorax are highly dependent on anatomical region and species. Modulus gradation, but not mean modulus, differed between the two types of fliers. In some regions, A. mellifera had a positive linear modulus gradient from cuticle interior to exterior of about 2 GPa. In M. sexta, the modulus gradients were variable and were not well represented by linear fits with respect to cuticle thickness. We utilized finite element modeling to assess how measured modulus gradients influenced maximum stress in cuticle. Stress was reduced when cuticle with a linear gradient was compressed from the high modulus side. These results support the protective role of the A. mellifera thorax cuticle. Our multi-faceted assessment advances our understanding of thorax cuticle structural and material heterogeneity and the potential benefit of material gradation to flying insects. Graphical Abstract O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY Statement of SignificanceThe insect thorax is essential for efficient flight but questions remain about the contribution of exoskeletal cuticle. We assessed the thorax cuticle using a high resolution multi-faceted approach to determine how cuticle properties vary within thorax anatomical regions and between fliers with asynchronous (honey bee; Apis mellifera) and synchronous (hawkmoth; Manduca sexta) muscles. We examined structure using histological staining, modulus using nanoindentation, and material composition using confocal scanning light microscopy. We further utilized finite element modeling to understand the effect of the modulus gradations observed experimentally on stress accumulation. Cuticle properties vary through cuticle thickness, by thorax region, and between flight lineages.