Jean-Christophe SANDOZ | Centre National de la Recherche Scientifique / French National Centre for Scientific Research (original) (raw)
Papers by Jean-Christophe SANDOZ
Neurobiology of Learning and Memory
Royal Society Open Science
Learning is a widespread phenomenon that allows behavioural flexibility when individuals face new... more Learning is a widespread phenomenon that allows behavioural flexibility when individuals face new situations. However, learned information may lose its value over time. If such a memory endures, it can be deleterious to individuals. The process of extinction allows memory updating when the initial information is not relevant anymore. Extinction is widespread among animals, including humans. We investigated associative appetitive learning in an ant species that is widely distributed in the Northern Hemisphere, Formica fusca . We studied acquisition and memory between 1 h and one week after conditioning, as well as the extinction process. Ants learn very rapidly, their memory lasts up to 3 days, decreases slowly over time and is highly resistant to extinction, even after a single conditioning trial. Using a pharmacological approach, we show that this single-trial memory critically depends on protein synthesis (long-term memory). These results indicate that individual ant workers of F....
Journal of Comparative Neurology
The Journal of comparative neurology, Jan 8, 2018
Insect mating behavior is controlled by a diverse array of sex-specific traits and strategies tha... more Insect mating behavior is controlled by a diverse array of sex-specific traits and strategies that evolved to maximize mating success. Orchid bees exhibit a unique suite of perfume-mediated mating behaviors. Male bees collect volatile compounds from their environment to concoct species-specific perfume mixtures that are presumably used to attract conspecific females. Despite a growing understanding of the ecology and evolution of chemical signaling in orchid bees, many aspects of the functional adaptations involved, in particular regarding sensory systems, remain unknown. Here we investigated male and female brain morphology in the common orchid bee Euglossa dilemma Bembé & Eltz. Males exhibited increased relative volumes of the Medulla, a visual brain region, which correlated with larger compound eye size (area). While the overall volume of olfactory brain regions was similar between sexes, the antennal lobes exhibited several sex-specific structures including one male-specific mac...
Neurobiology of learning and memory, Jan 21, 2018
The honey bee Apis mellifera is a major insect model for studying visual cognition. Free-flying h... more The honey bee Apis mellifera is a major insect model for studying visual cognition. Free-flying honey bees learn to associate different visual cues with a sucrose reward and may deploy sophisticated cognitive strategies to this end. Yet, the neural bases of these capacities cannot be studied in flying insects. Conversely, immobilized bees are accessible to neurobiological investigation but training them to respond appetitively to visual stimuli paired with sucrose reward is difficult. Here we succeeded in coupling visual conditioning in harnessed bees with pharmacological analyses on the role of octopamine (OA), dopamine (DA) and serotonin (5-HT) in visual learning. We also studied if and how these biogenic amines modulate sucrose responsiveness and phototaxis behaviour as intact reward and visual perception are essential prerequisites for appetitive visual learning. Our results suggest that both octopaminergic and dopaminergic signaling mediate either the appetitive sucrose signali...
Scientific reports, Jan 10, 2017
Free-flying honeybees exhibit remarkable cognitive capacities but the neural underpinnings of the... more Free-flying honeybees exhibit remarkable cognitive capacities but the neural underpinnings of these capacities cannot be studied in flying insects. Conversely, immobilized bees are accessible to neurobiological investigation but display poor visual learning. To overcome this limitation, we aimed at establishing a controlled visual environment in which tethered bees walking on a spherical treadmill learn to discriminate visual stimuli video projected in front of them. Freely flying bees trained to walk into a miniature Y-maze displaying these stimuli in a dark environment learned the visual discrimination efficiently when one of them (CS+) was paired with sucrose and the other with quinine solution (CS-). Adapting this discrimination to the treadmill paradigm with a tethered, walking bee was successful as bees exhibited robust discrimination and preferred the CS+ to the CS- after training. As learning was better in the maze, movement freedom, active vision and behavioral context migh...
Frontiers in Neuroanatomy
Eusocial Hymenoptera colonies are characterized by the presence of altruistic individuals, which ... more Eusocial Hymenoptera colonies are characterized by the presence of altruistic individuals, which rear their siblings instead of their own offspring. In the course of evolution, such sterile castes are thought to have emerged through the process of kin selection, altruistic traits being transmitted to following generation if they benefit relatives. By allowing kinship recognition, the detection of cuticular hydrocarbons (CHCs) might be instrumental for kin selection. In carpenter ants, a female-specific olfactory subsystem processes CHC information through antennal detection by basiconic sensilla. It is still unclear if other families of eusocial Hymenoptera use the same subsystem for sensing CHCs. Here, we examined the existence of such a subsystem in Vespidae (using the hornet Vespa velutina), a family in which eusociality emerged independently of ants. The antennae of both males and female hornets contain large basiconic sensilla. Sensory neurons from the large basiconic sensilla exclusively project to a conspicuous cluster of small glomeruli in the antennal lobe, with anatomical and immunoreactive features that are strikingly similar to those of the ant CHC-sensitive subsystem. Extracellular electrophysiological recordings further show that sensory neurons within hornet basiconic sensilla preferentially respond to CHCs. Although this subsystem is not female-specific in hornets, the observed similarities with the olfactory system of ants are striking. They suggest that the basiconic sensilla subsystem could be an ancestral trait, which may have played a key role in the advent of eusociality in these hymenopteran families by allowing kin recognition and the production of altruistic behaviors toward relatives.
Proceedings of the National Academy of Sciences
Scientific Reports
Although the honeybee is a crucial agricultural agent and a prominent scientific model organism, ... more Although the honeybee is a crucial agricultural agent and a prominent scientific model organism, crucial aspects of its reproductive behaviour are still unknown. During the mating season, honeybee males, the drones, gather in congregations 10-40 m above ground. Converging evidence suggests that drones emit a pheromone that can attract other drones, thereby increasing the size of the congregation. Virgin queens join the vicinity of the congregation after it has formed, and mate with as many as 20 males in mid-air. It is still unclear which sensory cues help virgin queens find drone congregations in the first place. Beside visual cues for long-range orientation, queens may use olfactory cues. We thus tested virgin queens' olfactory orientation on a walking simulator in which they have full control over odour stimulation. We show that sexually-mature virgin queens are attracted to the odour bouquet from a group of living drones. They are not attracted to the bouquet from a group of workers. In addition, nonsexually receptive females (workers) of the same age are not attracted to the drone odour bouquet. Interpreted in the context of mating, these results may suggest that virgin queens use volatile olfactory cues from the drones to find the congregations. Bees (Hymenoptera: Apoidea: Anthophila) represent more than 20,000 species, from solitary to eusocial, which display a wide spectrum of mating behaviours 1-3. Some species mate at the female emergence site (e.g. Colletes cunicularius 4 and Centris pallida 5), others at resource-based sites (Anthidium manicatum 6), while still others use nuptial flyways (Apis species 7). Despite this diversity of mating strategies, many bee species are known to use sex pheromones for reproduction 2. Sex pheromones are defined as odours, produced by either males or females, which stimulate behavioural reactions and/or induce physiological effects in the opposite sex, bringing the sexes together for the purpose of mating 2. Bees have large glandular systems allowing both females and males to produce sex pheromones which attract conspecifics. Female exocrine secretions from the mandibular glands are the most common bee sex pheromones. They induce male attraction in some Colletidae 4 , Andrenidae 8 and Apidae like carpenter bees 9 , stingless bees 10 , bumble bees 11 and honey bees 12. In some cases, the male secretions produced by the mandibular glands or the labial glands are also used to mark spots along male flight paths. These spots are attractive to females but also to conspecific males in some Andrenidae 13 , and Apidae, like carpenter bees 9 , bumble bees 14 and orchid bees 15. The importance of male-produced pheromones for bee reproduction has long been underestimated, probably because they were less intensively studied. Even in some well-studied species their existence and role are still unclear. A good example is the honeybee Apis mellifera, a worldwide economically valuable pollinator and a mainstream scientific model in diverse fields such as genetics, physiology, ethology, neurobiology and animal cognition 16-23. Honeybees are eusocial insects, characterized by a reproductive division of labor between one fertile female, the queen and thousands of facultatively sterile females, the workers 17. Honeybees display a particularly striking mating behaviour 24-30. During the mating season, and on favourable weather conditions, honeybee males, the drones, fly out and gather high in the air at discrete congregation areas located usually 10-40 m above ground, with a diameter of 30-200 m 7, 31-33. These drone congregations can contain as many as 11,000 drones from up to 240 different colonies 33-36. Then, about one hour after the peak of drones' departure, virgin queens leave the hive and fly to the vicinity of the drone congregation 7, 25, 37. As soon as a virgin queen is present, many drones are attracted to her, both by olfactory signals (the mandibular gland sex pheromone, 9-oxo-2-decenoic acid, 9-ODA) and by visual cues at shorter range 12, 38. Drones follow the virgin queen in a comet-like swarm and engage in a scramble competition, each individual struggling for the most promising position to approach and
Pest Management Science
Azadirachtin is a prominent natural pesticide and represents an alternative to conventional insec... more Azadirachtin is a prominent natural pesticide and represents an alternative to conventional insecticides. It has been successfully used against insect pests. However, its effects on reproduction require further analysis. Here we investigated lethal and sublethal effects of azadirachtin, on treated adults in a model insect, Drosophila melanogaster (Meigen). Dose-mortality relationships as well as several parameters of reproduction (mating, spermatogenesis, oogenesis and fertility) were examined. Neem-Azal, a commercial formulation of azadirachtin, applied topically on newly emerged adults, increased mortality with a positive dose-dependent relationship. The LD50 (0.63 μg) was determined 24 h after treatment using a non-linear regression. Adults surviving this dose had a mating success that was divided by 3 and a progeny production reduced by half when males were treated, and even more when females were treated. When combining probability of survival, of mating and reduced progeny, it appeared that LD50 induced a 98% reduction in reproductive rates. Reduced progeny was partially explained by the effect of adult treatment on gametes number and abnormalities. The number of cysts and the apical nuclei positions within the cysts decreased by 29.7% and 20%, respectively, in males. In females, the number of oocytes per ovary and the volume of basal oocytes also decreased by 16.1% and 32.4%, respectively. Azadirachtin causes significant toxic effects in both sexes and decreases the fecundity and fertility of D. melanogaster. Females are more sensitive to azadirachtin. © 2017 Society of Chemical Industry.
Handbook of Behavioral Neuroscience, 2013
Honeybee Neurobiology and Behavior, 2011
Journal of Experimental Biology, 2014
During the mating season, honeybee males, the drones, gather in congregation areas 10-40 m above ... more During the mating season, honeybee males, the drones, gather in congregation areas 10-40 m above ground. When a receptive female, a queen, enters the congregation, drones are attracted to her by queen-produced pheromones and visual cues and attempt to mate with the queen in mid-air. It is still unclear how drones and queens find the congregations. Visual cues on the horizon are most probably used for long-range orientation. For shorter-range orientation, however, attraction by a drone-produced aggregation pheromone has been proposed, yet so far its existence has not been confirmed conclusively. The low accessibility of congregation areas high up in the air is a major hurdle and precise control of experimental conditions often remains unsatisfactory in field studies. Here, we used a locomotion compensator-based walking simulator to investigate drones' innate odor preferences under controlled laboratory conditions. We tested behavioral responses of drones to 9-oxo-2-decenoic acid (9-ODA), the major queen-produced sexual attractant, and to queen mandibular pheromone (QMP), an artificial blend of 9-ODA and several other queen-derived components. While 9-ODA strongly dominates the odor bouquet of virgin queens, QMP rather resembles the bouquet of mated queens. In our assay, drones were attracted by 9-ODA, but not by QMP. We also investigated the potential attractiveness of male-derived odors by testing drones' orientation responses to the odor bouquet of groups of 10 living drones or workers. Our results demonstrate that honeybee drones are attracted by groups of other drones (but not by workers), which may indicate a role of drone-emitted cues for the formation of congregations.
Journal of Comparative Neurology, 2016
In the course of evolution, eusociality has appeared several times independently in Hymenoptera, ... more In the course of evolution, eusociality has appeared several times independently in Hymenoptera, within different families such as Apidae (bees), Formicidae (ants), and Vespidae (wasps and hornets), among others. The complex social organization of eusocial Hymenoptera relies on sophisticated olfactory communication systems. Whereas the olfactory systems of several bee and ant species have been well characterized, very little information is as yet available in Vespidae, although this family represents a highly successful insect group, displaying a wide range of life styles from solitary to eusocial. Using fluorescent labeling, confocal microscopy, and 3D reconstructions, we investigated the organization of the olfactory pathway in queens, workers, and males of the eusocial hornet Vespa velutina. First, we found that caste and sex dimorphism is weakly pronounced in hornets, with regard to both whole-brain morphology and antennal lobe organization, although several male-specific macroglomeruli are present. The V. velutina antennal lobe contains approximately 265 glomeruli (in females), grouped in nine conspicuous clusters formed by afferent tract subdivisions. As in bees and ants, hornets display a dual olfactory pathway, with two major efferent tracts, the medial and the lateral antennal lobe tracts (m- and l-ALT), separately arborizing two antennal lobe hemilobes and projecting to partially different regions of higher order olfactory centers. Finally, we found remarkable anatomical similarities in the glomerular cluster organizations among hornets, ants, and bees, suggesting the possible existence of homologies in the olfactory pathways of these eusocial Hymenoptera. We propose a common framework for describing AL compartmentalization across Hymenoptera and discuss possible evolutionary scenarios. J. Comp. Neurol. 524:2335-2359, 2016. © 2016 Wiley Periodicals, Inc.
Frontiers in Physiology, 2015
The recent development of the olfactory conditioning of the sting extension response (SER) has pr... more The recent development of the olfactory conditioning of the sting extension response (SER) has provided new insights into the mechanisms of aversive learning in honeybees. Until now, very little information has been gained concerning US detection and perception. In the initial version of SER conditioning, bees learned to associate an odor CS with an electric shock US. Recently, we proposed a modified version of SER conditioning, in which thermal stimulation with a heated probe is used as US. This procedure has the advantage of allowing topical US applications virtually everywhere on the honeybee body. In this study, we made use of this possibility and mapped thermal responsiveness on the honeybee body, by measuring workers' SER after applying heat on 41 different structures. We then show that bees can learn the CS-US association even when the heat US is applied on body structures that are not prominent sensory organs, here the vertex (back of the head) and the ventral abdomen. Next, we used a neuropharmalogical approach to evaluate the potential role of a recently described Transient Receptor Potential (TRP) channel, HsTRPA, on peripheral heat detection by bees. First, we applied HsTRPA activators to assess if such activation is sufficient for triggering SER. Second, we injected HsTRPA inhibitors to ask whether interfering with this TRP channel affects SER triggered by heat. These experiments suggest that HsTRPA may be involved in heat detection by bees, and represent a potential peripheral detection system in thermal SER conditioning.
Frontiers in Integrative Neuroscience, 2016
The function of parallel neural processing is a fundamental problem in Neuroscience, as it is fou... more The function of parallel neural processing is a fundamental problem in Neuroscience, as it is found across sensory modalities and evolutionary lineages, from insects to humans. Recently, parallel processing has attracted increased attention in the olfactory domain, with the demonstration in both insects and mammals that different populations of second-order neurons encode and/or process odorant information differently. Among insects, Hymenoptera present a striking olfactory system with a clear neural dichotomy from the periphery to higher-order centers, based on two main tracts of second-order (projection) neurons: the medial and lateral antennal lobe tracts (m-ALT and l-ALT). To unravel the functional role of these two pathways, we combined specific lesions of the m-ALT tract with behavioral experiments, using the classical conditioning of the proboscis extension response (PER conditioning). Lesioned and intact bees had to learn to associate an odorant (1-nonanol) with sucrose. Then the bees were subjected to a generalization procedure with a range of odorants differing in terms of their carbon chain length or functional group. We show that m-ALT lesion strongly affects acquisition of an odor-sucrose association. However, lesioned bees that still learned the association showed a normal gradient of decreasing generalization responses to increasingly dissimilar odorants. Generalization responses could be predicted to some extent by in vivo calcium imaging recordings of l-ALT neurons. The m-ALT pathway therefore seems necessary for normal classical olfactory conditioning performance.
PLOS ONE, 2015
The toxicity of pesticides used in agriculture towards non-targeted organisms and especially poll... more The toxicity of pesticides used in agriculture towards non-targeted organisms and especially pollinators has recently drawn the attention from a broad scientific community. Increased honeybee mortality observed worldwide certainly contributes to this interest. The potential role of several neurotoxic insecticides in triggering or potentiating honeybee mortality was considered, in particular phenylpyrazoles and neonicotinoids, given that they are widely used and highly toxic for insects. Along with their ability to kill insects at lethal doses, they can compromise survival at sublethal doses by producing subtle deleterious effects. In this study, we compared the bee's locomotor ability, which is crucial for many tasks within the hive (e.g. cleaning brood cells, feeding larvae…), before and after an acute sublethal exposure to one insecticide belonging to the two insecticide classes, fipronil and thiamethoxam. Additionally, we examined the locomotor ability after exposure to pyrethroids, an older chemical insecticide class still widely used and known to be highly toxic to bees as well. Our study focused on young bees (day 1 after emergence) since (i) few studies are available on locomotion at this stage and (ii) in recent years, pesticides have been reported to accumulate in different hive matrices, where young bees undergo their early development. At sublethal doses (SLD48h, i.e. causing no mortality at 48h), three pyrethroids, namely cypermethrin (2.5 ng/bee), tetramethrin (70 ng/bee), tau-fluvalinate (33 ng/bee) and the neonicotinoid thiamethoxam (3.8 ng/bee) caused a locomotor deficit in honeybees. While the SLD48h of fipronil (a phenylpyrazole, 0.5 ng/bee) had no measurable effect on locomotion, we observed high mortality several days after exposure, an effect that was not observed with the other insecticides. Although locomotor deficits observed in the sublethal range of pyrethroids and thiamethoxam would suggest deleterious effects in the field, the case of fipronil demonstrates that toxicity evaluation requires information on multiple endpoints (e.g. long term survival) to fully address pesticides risks for honeybees. Pyrethroid-induced locomotor deficits are discussed in light of recent advances regarding their mode of action on honeybee ion channels and current structure-function studies.
Learning & memory (Cold Spring Harbor, N.Y.), 2015
In honeybees, two olfactory conditioning protocols allow the study of appetitive and aversive Pav... more In honeybees, two olfactory conditioning protocols allow the study of appetitive and aversive Pavlovian associations. Appetitive conditioning of the proboscis extension response (PER) involves associating an odor, the conditioned stimulus (CS) with a sucrose solution, the unconditioned stimulus (US). Conversely, aversive conditioning of the sting extension response (SER) involves associating the odor CS with an electric or thermal shock US. Each protocol is based on the measure of a different behavioral response (proboscis versus sting) and both only provide binary responses (extension or not of the proboscis or sting). These limitations render the measure of the acquired valence of an odor CS difficult without testing the animals in a freely moving situation. Here, we studied the effects of both olfactory conditioning protocols on the movements of the antennae, which are crucial sensory organs for bees. As bees' antennae are highly mobile, we asked whether their movements in re...
![Research paper thumbnail of [Olfactory perception and learning in the honey bee (Apis mellifera): calcium imaging in the antenna lobe]](https://attachments.academia-assets.com/48117124/thumbnails/1.jpg)
](Journal de la Société de Biologie
Comptes Rendus de l Académie des Sciences - Series III - Sciences de la Vie
Olfactory learning in the honeybee was investigated using the conditionedproboscis extension refl... more Olfactory learning in the honeybee was investigated using the conditionedproboscis extension reflex on restrained individuals. We compared, under the same expérimental conditions, the most commonly used conditioning procédures, i.e. / trial, 3 massed trials (1 min inter-trial intervais), and 3 spaced trials (10 min inter-trial intervais) procédures, using linalool as the conditioned stimulus. Two experiments were performed in which worker bées were subjected to: (1) a single test at différent times (30 s to 14 dàys) after the conditioning procédure; (2) a first test within 3 h after the conditioning procédure, and were then retested daily (up to 5 tests). The memory trace of a learnt odorant stimulus could last for the lifetime of the bee, even after a single association with sugar.
Neurobiology of Learning and Memory
Royal Society Open Science
Learning is a widespread phenomenon that allows behavioural flexibility when individuals face new... more Learning is a widespread phenomenon that allows behavioural flexibility when individuals face new situations. However, learned information may lose its value over time. If such a memory endures, it can be deleterious to individuals. The process of extinction allows memory updating when the initial information is not relevant anymore. Extinction is widespread among animals, including humans. We investigated associative appetitive learning in an ant species that is widely distributed in the Northern Hemisphere, Formica fusca . We studied acquisition and memory between 1 h and one week after conditioning, as well as the extinction process. Ants learn very rapidly, their memory lasts up to 3 days, decreases slowly over time and is highly resistant to extinction, even after a single conditioning trial. Using a pharmacological approach, we show that this single-trial memory critically depends on protein synthesis (long-term memory). These results indicate that individual ant workers of F....
Journal of Comparative Neurology
The Journal of comparative neurology, Jan 8, 2018
Insect mating behavior is controlled by a diverse array of sex-specific traits and strategies tha... more Insect mating behavior is controlled by a diverse array of sex-specific traits and strategies that evolved to maximize mating success. Orchid bees exhibit a unique suite of perfume-mediated mating behaviors. Male bees collect volatile compounds from their environment to concoct species-specific perfume mixtures that are presumably used to attract conspecific females. Despite a growing understanding of the ecology and evolution of chemical signaling in orchid bees, many aspects of the functional adaptations involved, in particular regarding sensory systems, remain unknown. Here we investigated male and female brain morphology in the common orchid bee Euglossa dilemma Bembé & Eltz. Males exhibited increased relative volumes of the Medulla, a visual brain region, which correlated with larger compound eye size (area). While the overall volume of olfactory brain regions was similar between sexes, the antennal lobes exhibited several sex-specific structures including one male-specific mac...
Neurobiology of learning and memory, Jan 21, 2018
The honey bee Apis mellifera is a major insect model for studying visual cognition. Free-flying h... more The honey bee Apis mellifera is a major insect model for studying visual cognition. Free-flying honey bees learn to associate different visual cues with a sucrose reward and may deploy sophisticated cognitive strategies to this end. Yet, the neural bases of these capacities cannot be studied in flying insects. Conversely, immobilized bees are accessible to neurobiological investigation but training them to respond appetitively to visual stimuli paired with sucrose reward is difficult. Here we succeeded in coupling visual conditioning in harnessed bees with pharmacological analyses on the role of octopamine (OA), dopamine (DA) and serotonin (5-HT) in visual learning. We also studied if and how these biogenic amines modulate sucrose responsiveness and phototaxis behaviour as intact reward and visual perception are essential prerequisites for appetitive visual learning. Our results suggest that both octopaminergic and dopaminergic signaling mediate either the appetitive sucrose signali...
Scientific reports, Jan 10, 2017
Free-flying honeybees exhibit remarkable cognitive capacities but the neural underpinnings of the... more Free-flying honeybees exhibit remarkable cognitive capacities but the neural underpinnings of these capacities cannot be studied in flying insects. Conversely, immobilized bees are accessible to neurobiological investigation but display poor visual learning. To overcome this limitation, we aimed at establishing a controlled visual environment in which tethered bees walking on a spherical treadmill learn to discriminate visual stimuli video projected in front of them. Freely flying bees trained to walk into a miniature Y-maze displaying these stimuli in a dark environment learned the visual discrimination efficiently when one of them (CS+) was paired with sucrose and the other with quinine solution (CS-). Adapting this discrimination to the treadmill paradigm with a tethered, walking bee was successful as bees exhibited robust discrimination and preferred the CS+ to the CS- after training. As learning was better in the maze, movement freedom, active vision and behavioral context migh...
Frontiers in Neuroanatomy
Eusocial Hymenoptera colonies are characterized by the presence of altruistic individuals, which ... more Eusocial Hymenoptera colonies are characterized by the presence of altruistic individuals, which rear their siblings instead of their own offspring. In the course of evolution, such sterile castes are thought to have emerged through the process of kin selection, altruistic traits being transmitted to following generation if they benefit relatives. By allowing kinship recognition, the detection of cuticular hydrocarbons (CHCs) might be instrumental for kin selection. In carpenter ants, a female-specific olfactory subsystem processes CHC information through antennal detection by basiconic sensilla. It is still unclear if other families of eusocial Hymenoptera use the same subsystem for sensing CHCs. Here, we examined the existence of such a subsystem in Vespidae (using the hornet Vespa velutina), a family in which eusociality emerged independently of ants. The antennae of both males and female hornets contain large basiconic sensilla. Sensory neurons from the large basiconic sensilla exclusively project to a conspicuous cluster of small glomeruli in the antennal lobe, with anatomical and immunoreactive features that are strikingly similar to those of the ant CHC-sensitive subsystem. Extracellular electrophysiological recordings further show that sensory neurons within hornet basiconic sensilla preferentially respond to CHCs. Although this subsystem is not female-specific in hornets, the observed similarities with the olfactory system of ants are striking. They suggest that the basiconic sensilla subsystem could be an ancestral trait, which may have played a key role in the advent of eusociality in these hymenopteran families by allowing kin recognition and the production of altruistic behaviors toward relatives.
Proceedings of the National Academy of Sciences
Scientific Reports
Although the honeybee is a crucial agricultural agent and a prominent scientific model organism, ... more Although the honeybee is a crucial agricultural agent and a prominent scientific model organism, crucial aspects of its reproductive behaviour are still unknown. During the mating season, honeybee males, the drones, gather in congregations 10-40 m above ground. Converging evidence suggests that drones emit a pheromone that can attract other drones, thereby increasing the size of the congregation. Virgin queens join the vicinity of the congregation after it has formed, and mate with as many as 20 males in mid-air. It is still unclear which sensory cues help virgin queens find drone congregations in the first place. Beside visual cues for long-range orientation, queens may use olfactory cues. We thus tested virgin queens' olfactory orientation on a walking simulator in which they have full control over odour stimulation. We show that sexually-mature virgin queens are attracted to the odour bouquet from a group of living drones. They are not attracted to the bouquet from a group of workers. In addition, nonsexually receptive females (workers) of the same age are not attracted to the drone odour bouquet. Interpreted in the context of mating, these results may suggest that virgin queens use volatile olfactory cues from the drones to find the congregations. Bees (Hymenoptera: Apoidea: Anthophila) represent more than 20,000 species, from solitary to eusocial, which display a wide spectrum of mating behaviours 1-3. Some species mate at the female emergence site (e.g. Colletes cunicularius 4 and Centris pallida 5), others at resource-based sites (Anthidium manicatum 6), while still others use nuptial flyways (Apis species 7). Despite this diversity of mating strategies, many bee species are known to use sex pheromones for reproduction 2. Sex pheromones are defined as odours, produced by either males or females, which stimulate behavioural reactions and/or induce physiological effects in the opposite sex, bringing the sexes together for the purpose of mating 2. Bees have large glandular systems allowing both females and males to produce sex pheromones which attract conspecifics. Female exocrine secretions from the mandibular glands are the most common bee sex pheromones. They induce male attraction in some Colletidae 4 , Andrenidae 8 and Apidae like carpenter bees 9 , stingless bees 10 , bumble bees 11 and honey bees 12. In some cases, the male secretions produced by the mandibular glands or the labial glands are also used to mark spots along male flight paths. These spots are attractive to females but also to conspecific males in some Andrenidae 13 , and Apidae, like carpenter bees 9 , bumble bees 14 and orchid bees 15. The importance of male-produced pheromones for bee reproduction has long been underestimated, probably because they were less intensively studied. Even in some well-studied species their existence and role are still unclear. A good example is the honeybee Apis mellifera, a worldwide economically valuable pollinator and a mainstream scientific model in diverse fields such as genetics, physiology, ethology, neurobiology and animal cognition 16-23. Honeybees are eusocial insects, characterized by a reproductive division of labor between one fertile female, the queen and thousands of facultatively sterile females, the workers 17. Honeybees display a particularly striking mating behaviour 24-30. During the mating season, and on favourable weather conditions, honeybee males, the drones, fly out and gather high in the air at discrete congregation areas located usually 10-40 m above ground, with a diameter of 30-200 m 7, 31-33. These drone congregations can contain as many as 11,000 drones from up to 240 different colonies 33-36. Then, about one hour after the peak of drones' departure, virgin queens leave the hive and fly to the vicinity of the drone congregation 7, 25, 37. As soon as a virgin queen is present, many drones are attracted to her, both by olfactory signals (the mandibular gland sex pheromone, 9-oxo-2-decenoic acid, 9-ODA) and by visual cues at shorter range 12, 38. Drones follow the virgin queen in a comet-like swarm and engage in a scramble competition, each individual struggling for the most promising position to approach and
Pest Management Science
Azadirachtin is a prominent natural pesticide and represents an alternative to conventional insec... more Azadirachtin is a prominent natural pesticide and represents an alternative to conventional insecticides. It has been successfully used against insect pests. However, its effects on reproduction require further analysis. Here we investigated lethal and sublethal effects of azadirachtin, on treated adults in a model insect, Drosophila melanogaster (Meigen). Dose-mortality relationships as well as several parameters of reproduction (mating, spermatogenesis, oogenesis and fertility) were examined. Neem-Azal, a commercial formulation of azadirachtin, applied topically on newly emerged adults, increased mortality with a positive dose-dependent relationship. The LD50 (0.63 μg) was determined 24 h after treatment using a non-linear regression. Adults surviving this dose had a mating success that was divided by 3 and a progeny production reduced by half when males were treated, and even more when females were treated. When combining probability of survival, of mating and reduced progeny, it appeared that LD50 induced a 98% reduction in reproductive rates. Reduced progeny was partially explained by the effect of adult treatment on gametes number and abnormalities. The number of cysts and the apical nuclei positions within the cysts decreased by 29.7% and 20%, respectively, in males. In females, the number of oocytes per ovary and the volume of basal oocytes also decreased by 16.1% and 32.4%, respectively. Azadirachtin causes significant toxic effects in both sexes and decreases the fecundity and fertility of D. melanogaster. Females are more sensitive to azadirachtin. © 2017 Society of Chemical Industry.
Handbook of Behavioral Neuroscience, 2013
Honeybee Neurobiology and Behavior, 2011
Journal of Experimental Biology, 2014
During the mating season, honeybee males, the drones, gather in congregation areas 10-40 m above ... more During the mating season, honeybee males, the drones, gather in congregation areas 10-40 m above ground. When a receptive female, a queen, enters the congregation, drones are attracted to her by queen-produced pheromones and visual cues and attempt to mate with the queen in mid-air. It is still unclear how drones and queens find the congregations. Visual cues on the horizon are most probably used for long-range orientation. For shorter-range orientation, however, attraction by a drone-produced aggregation pheromone has been proposed, yet so far its existence has not been confirmed conclusively. The low accessibility of congregation areas high up in the air is a major hurdle and precise control of experimental conditions often remains unsatisfactory in field studies. Here, we used a locomotion compensator-based walking simulator to investigate drones' innate odor preferences under controlled laboratory conditions. We tested behavioral responses of drones to 9-oxo-2-decenoic acid (9-ODA), the major queen-produced sexual attractant, and to queen mandibular pheromone (QMP), an artificial blend of 9-ODA and several other queen-derived components. While 9-ODA strongly dominates the odor bouquet of virgin queens, QMP rather resembles the bouquet of mated queens. In our assay, drones were attracted by 9-ODA, but not by QMP. We also investigated the potential attractiveness of male-derived odors by testing drones' orientation responses to the odor bouquet of groups of 10 living drones or workers. Our results demonstrate that honeybee drones are attracted by groups of other drones (but not by workers), which may indicate a role of drone-emitted cues for the formation of congregations.
Journal of Comparative Neurology, 2016
In the course of evolution, eusociality has appeared several times independently in Hymenoptera, ... more In the course of evolution, eusociality has appeared several times independently in Hymenoptera, within different families such as Apidae (bees), Formicidae (ants), and Vespidae (wasps and hornets), among others. The complex social organization of eusocial Hymenoptera relies on sophisticated olfactory communication systems. Whereas the olfactory systems of several bee and ant species have been well characterized, very little information is as yet available in Vespidae, although this family represents a highly successful insect group, displaying a wide range of life styles from solitary to eusocial. Using fluorescent labeling, confocal microscopy, and 3D reconstructions, we investigated the organization of the olfactory pathway in queens, workers, and males of the eusocial hornet Vespa velutina. First, we found that caste and sex dimorphism is weakly pronounced in hornets, with regard to both whole-brain morphology and antennal lobe organization, although several male-specific macroglomeruli are present. The V. velutina antennal lobe contains approximately 265 glomeruli (in females), grouped in nine conspicuous clusters formed by afferent tract subdivisions. As in bees and ants, hornets display a dual olfactory pathway, with two major efferent tracts, the medial and the lateral antennal lobe tracts (m- and l-ALT), separately arborizing two antennal lobe hemilobes and projecting to partially different regions of higher order olfactory centers. Finally, we found remarkable anatomical similarities in the glomerular cluster organizations among hornets, ants, and bees, suggesting the possible existence of homologies in the olfactory pathways of these eusocial Hymenoptera. We propose a common framework for describing AL compartmentalization across Hymenoptera and discuss possible evolutionary scenarios. J. Comp. Neurol. 524:2335-2359, 2016. © 2016 Wiley Periodicals, Inc.
Frontiers in Physiology, 2015
The recent development of the olfactory conditioning of the sting extension response (SER) has pr... more The recent development of the olfactory conditioning of the sting extension response (SER) has provided new insights into the mechanisms of aversive learning in honeybees. Until now, very little information has been gained concerning US detection and perception. In the initial version of SER conditioning, bees learned to associate an odor CS with an electric shock US. Recently, we proposed a modified version of SER conditioning, in which thermal stimulation with a heated probe is used as US. This procedure has the advantage of allowing topical US applications virtually everywhere on the honeybee body. In this study, we made use of this possibility and mapped thermal responsiveness on the honeybee body, by measuring workers' SER after applying heat on 41 different structures. We then show that bees can learn the CS-US association even when the heat US is applied on body structures that are not prominent sensory organs, here the vertex (back of the head) and the ventral abdomen. Next, we used a neuropharmalogical approach to evaluate the potential role of a recently described Transient Receptor Potential (TRP) channel, HsTRPA, on peripheral heat detection by bees. First, we applied HsTRPA activators to assess if such activation is sufficient for triggering SER. Second, we injected HsTRPA inhibitors to ask whether interfering with this TRP channel affects SER triggered by heat. These experiments suggest that HsTRPA may be involved in heat detection by bees, and represent a potential peripheral detection system in thermal SER conditioning.
Frontiers in Integrative Neuroscience, 2016
The function of parallel neural processing is a fundamental problem in Neuroscience, as it is fou... more The function of parallel neural processing is a fundamental problem in Neuroscience, as it is found across sensory modalities and evolutionary lineages, from insects to humans. Recently, parallel processing has attracted increased attention in the olfactory domain, with the demonstration in both insects and mammals that different populations of second-order neurons encode and/or process odorant information differently. Among insects, Hymenoptera present a striking olfactory system with a clear neural dichotomy from the periphery to higher-order centers, based on two main tracts of second-order (projection) neurons: the medial and lateral antennal lobe tracts (m-ALT and l-ALT). To unravel the functional role of these two pathways, we combined specific lesions of the m-ALT tract with behavioral experiments, using the classical conditioning of the proboscis extension response (PER conditioning). Lesioned and intact bees had to learn to associate an odorant (1-nonanol) with sucrose. Then the bees were subjected to a generalization procedure with a range of odorants differing in terms of their carbon chain length or functional group. We show that m-ALT lesion strongly affects acquisition of an odor-sucrose association. However, lesioned bees that still learned the association showed a normal gradient of decreasing generalization responses to increasingly dissimilar odorants. Generalization responses could be predicted to some extent by in vivo calcium imaging recordings of l-ALT neurons. The m-ALT pathway therefore seems necessary for normal classical olfactory conditioning performance.
PLOS ONE, 2015
The toxicity of pesticides used in agriculture towards non-targeted organisms and especially poll... more The toxicity of pesticides used in agriculture towards non-targeted organisms and especially pollinators has recently drawn the attention from a broad scientific community. Increased honeybee mortality observed worldwide certainly contributes to this interest. The potential role of several neurotoxic insecticides in triggering or potentiating honeybee mortality was considered, in particular phenylpyrazoles and neonicotinoids, given that they are widely used and highly toxic for insects. Along with their ability to kill insects at lethal doses, they can compromise survival at sublethal doses by producing subtle deleterious effects. In this study, we compared the bee's locomotor ability, which is crucial for many tasks within the hive (e.g. cleaning brood cells, feeding larvae…), before and after an acute sublethal exposure to one insecticide belonging to the two insecticide classes, fipronil and thiamethoxam. Additionally, we examined the locomotor ability after exposure to pyrethroids, an older chemical insecticide class still widely used and known to be highly toxic to bees as well. Our study focused on young bees (day 1 after emergence) since (i) few studies are available on locomotion at this stage and (ii) in recent years, pesticides have been reported to accumulate in different hive matrices, where young bees undergo their early development. At sublethal doses (SLD48h, i.e. causing no mortality at 48h), three pyrethroids, namely cypermethrin (2.5 ng/bee), tetramethrin (70 ng/bee), tau-fluvalinate (33 ng/bee) and the neonicotinoid thiamethoxam (3.8 ng/bee) caused a locomotor deficit in honeybees. While the SLD48h of fipronil (a phenylpyrazole, 0.5 ng/bee) had no measurable effect on locomotion, we observed high mortality several days after exposure, an effect that was not observed with the other insecticides. Although locomotor deficits observed in the sublethal range of pyrethroids and thiamethoxam would suggest deleterious effects in the field, the case of fipronil demonstrates that toxicity evaluation requires information on multiple endpoints (e.g. long term survival) to fully address pesticides risks for honeybees. Pyrethroid-induced locomotor deficits are discussed in light of recent advances regarding their mode of action on honeybee ion channels and current structure-function studies.
Learning & memory (Cold Spring Harbor, N.Y.), 2015
In honeybees, two olfactory conditioning protocols allow the study of appetitive and aversive Pav... more In honeybees, two olfactory conditioning protocols allow the study of appetitive and aversive Pavlovian associations. Appetitive conditioning of the proboscis extension response (PER) involves associating an odor, the conditioned stimulus (CS) with a sucrose solution, the unconditioned stimulus (US). Conversely, aversive conditioning of the sting extension response (SER) involves associating the odor CS with an electric or thermal shock US. Each protocol is based on the measure of a different behavioral response (proboscis versus sting) and both only provide binary responses (extension or not of the proboscis or sting). These limitations render the measure of the acquired valence of an odor CS difficult without testing the animals in a freely moving situation. Here, we studied the effects of both olfactory conditioning protocols on the movements of the antennae, which are crucial sensory organs for bees. As bees' antennae are highly mobile, we asked whether their movements in re...
Journal de la Société de Biologie
Comptes Rendus de l Académie des Sciences - Series III - Sciences de la Vie
Olfactory learning in the honeybee was investigated using the conditionedproboscis extension refl... more Olfactory learning in the honeybee was investigated using the conditionedproboscis extension reflex on restrained individuals. We compared, under the same expérimental conditions, the most commonly used conditioning procédures, i.e. / trial, 3 massed trials (1 min inter-trial intervais), and 3 spaced trials (10 min inter-trial intervais) procédures, using linalool as the conditioned stimulus. Two experiments were performed in which worker bées were subjected to: (1) a single test at différent times (30 s to 14 dàys) after the conditioning procédure; (2) a first test within 3 h after the conditioning procédure, and were then retested daily (up to 5 tests). The memory trace of a learnt odorant stimulus could last for the lifetime of the bee, even after a single association with sugar.