Degradation of Pheromone and Plant Volatile Components by a Same Odorant-Degrading Enzyme in the Cotton Leafworm, Spodoptera littoralis

Background: Odorant-Degrading Enzymes (ODEs) are supposed to be involved in the signal inactivation step within the olfactory sensilla of insects by quickly removing odorant molecules from the vicinity of the olfactory receptors. Only three ODEs have been both identified at the molecular level and functionally characterized: two were specialized in the degradation of pheromone compounds and the last one was shown to degrade a plant odorant. Methodology: Previous work has shown that the antennae of the cotton leafworm Spodoptera littoralis , a worldwide pest of agricultural crops, express numerous candidate ODEs. We focused on an esterase overexpressed in males antennae, namely SlCXE7. We studied its expression patterns and tested its catalytic properties towards three odorants, i.e. the two female sex pheromone components and a green leaf volatile emitted by host plants. Conclusion: SlCXE7 expression was concomitant during development with male responsiveness to odorants and during adult scotophase with the period of male most active sexual behaviour. Furthermore, SlCXE7 transcription could be induced by male exposure to the main pheromone component, suggesting a role of Pheromone-Degrading Enzyme. Interestingly, recombinant SlCXE7 was able to efficiently hydrolyze the pheromone compounds but also the plant volatile, with a higher affinity for the pheromone than for the plant compound. In male antennae, SlCXE7 expression was associated with both long and short sensilla, tuned to sex pheromones or plant odours, respectively. Our results thus suggested that a same ODE could have a dual function depending of it sensillar localisation. Within the pheromone-sensitive sensilla, SlCXE7 may play a role in pheromone signal termination and in reduction of odorant background noise, whereas it could be involved in plant odorant inactivation within the short sensilla

Degradation of Pheromone and Plant Volatile Components by a Same Odorant-Degrading Enzyme in the Cotton Leafworm, Spodoptera littoralis

Background: Odorant-Degrading Enzymes (ODEs) are supposed to be involved in the signal inactivation step within the olfactory sensilla of insects by quickly removing odorant molecules from the vicinity of the olfactory receptors. Only three ODEs have been both identified at the molecular level and functionally characterized: two were specialized in the degradation of pheromone compounds and the last one was shown to degrade a plant odorant. Methodology: Previous work has shown that the antennae of the cotton leafworm Spodoptera littoralis , a worldwide pest of agricultural crops, express numerous candidate ODEs. We focused on an esterase overexpressed in males antennae, namely SlCXE7. We studied its expression patterns and tested its catalytic properties towards three odorants, i.e. the two female sex pheromone components and a green leaf volatile emitted by host plants. Conclusion: SlCXE7 expression was concomitant during development with male responsiveness to odorants and during adult scotophase with the period of male most active sexual behaviour. Furthermore, SlCXE7 transcription could be induced by male exposure to the main pheromone component, suggesting a role of Pheromone-Degrading Enzyme. Interestingly, recombinant SlCXE7 was able to efficiently hydrolyze the pheromone compounds but also the plant volatile, with a higher affinity for the pheromone than for the plant compound. In male antennae, SlCXE7 expression was associated with both long and short sensilla, tuned to sex pheromones or plant odours, respectively. Our results thus suggested that a same ODE could have a dual function depending of it sensillar localisation. Within the pheromone-sensitive sensilla, SlCXE7 may play a role in pheromone signal termination and in reduction of odorant background noise, whereas it could be involved in plant odorant inactivation within the short sensilla

Behavioral and metabolic effects of sublethal doses of two insecticides, chlorpyrifos and methomyl, in the Egyptian cotton leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae)

International audienceInsecticides have long been used as the main method in limiting agricultural pests, but their widespread use has resulted in environmental pollution, development of resistances, and biodiversity reduction. The effects of insecticides at low residual doses on both the targeted crop pest species and beneficial insects have become a major concern. In particular, these low doses can induce unexpected positive (hormetic) effects on pest insects, such as surges in population growth exceeding what would have been observed without pesticide application. Methomyl and chlorpyrifos are two insecticides commonly used to control the population levels of the cotton leafworm Spodoptera littoralis, a major pest moth. The aim of the present study was to examine the effects of sublethal doses of these two pesticides, known to present a residual activity and persistence in the environment, on the moth physiology. Using a metabolomic approach, we showed that sublethal doses of methomyl and chlorpyrifos have a systemic effect on the treated insects. We also demonstrated a behavioral disruption of S. littoralis larvae exposed to sublethal doses of methomyl, whereas no effects were observed for the same doses of chlorpyrifos. Interestingly, we highlighted that sublethal doses of both pesticides did not induce a change in acetylcholinesterase activity in head of exposed larva

Isolation and Maintenance-Free Culture of Contractile Myotubes from Manduca sexta Embryos

Skeletal muscle tissue engineering has the potential to treat tissue loss and degenerative diseases. However, these systems are also applicable for a variety of devices where actuation is needed, such as microelectromechanical systems (MEMS) and robotics. Most current efforts to generate muscle bioactuators are focused on using mammalian cells, which require exacting conditions for survival and function. In contrast, invertebrate cells are more environmentally robust, metabolically adaptable and relatively autonomous. Our hypothesis is that the use of invertebrate muscle cells will obviate many of the limitations encountered when mammalian cells are used for bioactuation. We focus on the tobacco hornworm, Manduca sexta, due to its easy availability, large size and well-characterized muscle contractile properties. Using isolated embryonic cells, we have developed culture conditions to grow and characterize contractile M. sexta muscles. The insect hormone 20-hydroxyecdysone was used to induce differentiation in the system, resulting in cells that stained positive for myosin, contract spontaneously for the duration of the culture, and do not require media changes over periods of more than a month. These cells proliferate under normal conditions, but the application of juvenile hormone induced further proliferation and inhibited differentiation. Cellular metabolism under normal and low glucose conditions was compared for C2C12 mouse and M. sexta myoblast cells. While differentiated C2C12 cells consumed glucose and produced lactate over one week as expected, M. sexta muscle did not consume significant glucose, and lactate production exceeded mammalian muscle production on a per cell basis. Contractile properties were evaluated using index of movement analysis, which demonstrated the potential of these cells to perform mechanical work. The ability of cultured M. sexta muscle to continuously function at ambient conditions without medium replenishment, combined with the interesting metabolic properties, suggests that this cell source is a promising candidate for further investigation toward bioactuator applications

Mécanismes moléculaires impliqués dans le contrôle de la prolifération cellulaire par les ecdysteroïdes chez le Lépidoptère Plodia interpunctella

PARIS-BIUSJ-Thèses (751052125) / SudocPARIS-BIUSJ-Physique recherche (751052113) / SudocSudocFranceF

Heat shock protein responses to salinity, food deprivation, and temperature in the invasive ground beetle Merizodus soledadinus at the Kerguelen Islands

International audienceThe ground beetle Merizodus soledadinus was introduced a century ago to the Kerguelen Islands. It has since become invasive and has colonized most coastlines east of this archipelago. In invaded intertidal zones, M. soledadinus has to deal with substrates that can reach high salinity levels. In addition to saline stress, the rapid spread of this invasive insect on the Kerguelen Islands may result in starving during dispersal, especially during winter periods. In order to gain a further understanding of the factors that have contributed to the success of this insect in invading the Kerguelen Islands, we assessed the variability in the expression of heat shock cognate 70 (HSC70) in M. soledadinus. HSC70 are constitutively expressed by insects, and we examined if the expression of HSC70 could picture the health degree of the ground beetles exposed at a range of environmental conditions, for example, varying temperatures (0, 4, 8, 12, and 20 C), trophic status (fed and food-deprived individuals), and saline conditions (salinities of 0, 35, and 70). We found that HSC70 expression decreased with increasing salinity. HSC70 expression was not modified in response to non-extreme thermal variations or short-term food deprivation, which did not appear to be stressful conditions for M. soledadinus given the survival results. We concluded that HSC70 expression may serve as a molecular indicator of the levels of well-being of this ground beetle when exposed to a range of environmental perturbations

Growth potential : effects of temperature on cellular proliferation at molecular levels in Plodia interpunctella (Lepidoptera : Pyralidae).

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Impact of cold-exposure and subsequent recovery on the differentiation of Lepidopteran cells at cellular and molecular levels.

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Effects of cold-exposure and subsequent recovery on cellular proliferation with influence of 20-hydroxyecdysone in a lepidopteran cell line (IAL-PID2).

International audienceIn developing insects, the peak level of 20-hydroxyecdysone (20E) initiates a decrease in cyclin expression, which subsequently triggers an arrest of cellular proliferation and the start of differentiation, finally culminating in the moult. We investigated the impact of cold-exposure (4 °C) and recovery (26 °C) on the cell cycle activity of the Plodia interpunctella Lepidoptera cell line IAL-PID2 and on the expression of B-type cyclin (PcycB), ecdysone receptor (B1-isoform; PiEcR-B1), and Hsc70 (PiHsc70) mRNA. Cold-exposure significantly reduced expression of these mRNAs, while their levels increased to above control values during subsequent recovery at the normal growth temperature. When cold-exposed cells were returned to 26 °C, cell cycle activity restarted, but apoptosis was strongly increased. The presence of 20E appeared to increase this apoptotic phenomenon. This result is consistent with the described protective role of 20E against a variety of stressors and with the capacity of 20E to induce cell death in different situations. Here, we illustrate for the first time a connection between 20E treatment and Hsc70 expression during cold-exposure and subsequent recovery in insect cells. Combined with the 20E-induced apoptotic response, our results suggest that regulation of Hsc70 expression by 20E could act in synergy with the control of apoptotic cell death in order to optimize the survival of specific cell populations after a period of cold-exposure