Etude des sous-unités auxiliaires du canal sodium dépendant du potentiel chez l’insecte : approches moléculaires, électrophysiologiques et pharmacologiques (Thèse de Doctorat d'Université)

Voltage-gated sodium (Nav) channel is a crucial molecular component of the cellular excitability. It represents a target of choice for neurotoxic insecticides used in pest control. Pyrazoline-type insecticide interacts with the main Nav channel subunit with a preference for its inactivated state. A recent study showed that auxiliary subunits of Drosophila melanogaster modified this conformation. However, little information is available concerning the role and the regulation of these auxiliary subunits. The objectives of this thesis were to characterize the auxiliary subunits of the American cockroach Periplaneta americana by molecular, electrophysiological and pharmacological approaches, in order to specify their functions. The first part of this work concerns the neuronal TEH1 subunit. Two variants, PaTEH1A and PaTEH1B resulting from an intron retention modifying only the C-terminal extremity, were cloned. Using the heterologous expression system Xenopus oocytes and the two microelectrodes voltage clamp technique, we highlighted that the C-terminal extremity was involved in the modulation of Nav channels electrophysiological and pharmacological properties. The second part concerns the discovery of other auxiliary subunits. We identified several variants resulting from alternative splicing events (2 variants for PaTipE and 4 for PaTEH2). Altogether, our results indicate that auxiliary subunits are diverse and play an important role in the modulation of Na+ current and should be considered to improve pharmacological studies

The effect of renovation of long-term temperate grassland on N2O emissions and N leaching from contrasting soils

pre-printRenovation of long-term grassland is associated with a peak in soil organic N mineralisation which, coupled with diminished plant N uptake can lead to large gaseous and leaching N losses. This study reports on the effect of ploughing and subsequent N fertilisation on the N2O emissions and DON/NO3− leaching, and evaluates the impact of ploughing technique on the magnitude and profile of N losses. This study was carried out on isolated grassland lysimeters of three Irish soils representing contrasting drainage properties (well-drained Clonakilty, moderately-drained Elton and poorly-drained Rathangan). Lysimeters were manually ploughed simulating conventional (CT) and minimum tillage (MT) as two treatments. Renovation of grassland increased N2O flux to a maximum of 0.9 kg N2O–N ha− 1 from poorly-drained soil over four days after treatment. Although there was no difference between CT and MT in the post-ploughing period, the treatment influenced subsequent N2O after fertiliser applications. Fertilisation remained the major driver of N losses therefore reducing fertilisation rate post-planting to account for N mineralised through grassland renovation could reduce the losses in medium to longer term. Leaching was a significant loss pathway, with the cumulative drainage volume and N leached highly influenced by soil type. Overall, the total N losses (N2O + N leached) were lowest from poorly and moderately draining soil and highest for the well draining soil, reflecting the dominance of leaching on total N losses and the paramount importance of soil properties

Anisotropy of the effective toughness of layered media

This continues the study of the effective toughness of layered materials started in Hossain et al. (2014) and Hsueh et al. (2018), with a focus on anisotropy. We use the phase-field model and the surfing boundary condition to propagate a crack macroscopically at various angles to the layers. We study two idealized situations, the first where the elastic modulus is uniform while the toughness alternates and a second where the toughness is uniform and the elastic modulus alternates. We find that in the first case of toughness heterogeneity the effective toughness displays ‘anomalous isotropy’ in that it is independent of the propagation direction and equal to that of the tougher material except when the crack propagation is parallel to the layers. In the second case of elastic heterogeneity, we find the behavior more anisotropic and consistent with the toughening effects of stress fluctuation and need for crack renucleation at the compliant-to-stiff interface. In both cases, the effective toughness is not convex in the sense of interfacial energy or Wulff shape reflecting the fact that crack propagation follows a critical path. Further, in both cases the crack path is not straight and consistent with a maximal dissipation principle. Finally, the effective toughness depends on the contrast and pinning, rather than on the extent of crack fluctuation

Stress fluctuation, crack renucleation and toughening in layered materials

It has been established that contrast in the elastic properties can lead to enhancement of fracture toughness in heterogeneous materials. Focussing on layered materials as a model system, we show that this enhancement is a result of two distinct phenomena – first, fluctuations in stress leading to regions where the stress intensity at the crack is considerably smaller than that of the macroscopically applied value; and second, the lack of stress intensity when a crack is at a compliant to stiff interface thereby requiring renucleation. Using theoretical, computational and experimental methods, we study two geometries – a layered material and a layered material with a narrow channel – to separate the two phenomena. The stress fluctuation is present in both, but renucleation is present only in the layered medium. We provide quantitative estimates for the enhanced toughness

A variational model for fracture and debonding of thin films under in-plane loadings

We study fracture and debonding of a thin stiff film bonded to a rigid substrate through a thin compliant layer, introducing a two-dimensional variational fracture model in brittle elasticity. Fractures are naturally distinguished between transverse cracks in the film (curves in 2D) and debonded surfaces (2D planar regions). In order to study the mechanical response of such systems under increasing loads, we formulate a dimension-reduced, rate-independent, irreversible evolution law accounting for both transverse fracture and debonding. We propose a numerical implementation based on a regularized formulation of the fracture problem via a gradient damage functional, and provide an illustration of its capabilities exploring complex crack patterns, showing a qualitative comparison with geometrically involved real life examples. Moreover, we justify the underlying dimension-reduced model in the setting of scalar-valued displacement fields by a rigorous asymptotic analysis using Γ-convergence, starting from the three-dimensional variational fracture (free-discontinuity) problem under precise scaling hypotheses on material and geometric parameters. © 2014 Elsevier Ltd