Julie Gaburro

Visual effects in great bowerbird sexual displays and their implications for signal design

It is often assumed that the primary purpose of a males sexual display is to provide information about quality, or to strongly stimulate prospective mates, but other functions of courtship displays have been relatively neglected. Male great bowerbirds (Ptilonorhynchus nuchalis) construct bowers that exploit the females predictable field of view (FOV) during courtship displays by creating forced perspective illusions, and the quality of illusion is a good predictor of mating success. Here, we present and discuss two additional components of male courtship displays that use the females predetermined viewpoint: (i) the rapid and diverse flashing of coloured objects within her FOV and (ii) chromatic adaptation of the females eyes that alters her perception of the colour of the displayed objects. Neither is directly related to mating success, but both are likely to increase signal efficacy, and may also be associated with attracting and holding the females attention. Signal efficacy is constrained by trade-offs between the signal components; there are both positive and negative interactions within multicomponent signals. Important signal components may have a threshold effect on fitness rather than the often assumed linear relationship.

Neurophysiology of Insects Using Microelectrode Arrays: Current Trends and Future Prospects

Simple to complex behaviors are directed by the brain, which possess nervous cells, called neurons. Mammals have billions of neurons, organized in networks, making their study difficult. Although methods have well evolved since the last century, studying a simpler model is the key to resolving neuronal communication. In this review, we demonstrate that insects are an excellent model and tool to understand neural mechanisms. Moreover, new technology, such as Microelectrodes Arrays (MEAs), is an innovative method which opens the possibility to study neuron clusters, rather than individual cells. A combined method of an insect model and MEAs technology may lead to great discoveries in neurophysiology, advancing progress in pharmacology, infectious and neurodegenerative diseases, agriculture maintenance and robotics.

Neurotropism and behavioral changes associated with Zika infection in the vector Aedes aegypti

Understanding Zika virus infection dynamics is essential, as its recent emergence revealed possible devastating neuropathologies in humans, thus causing a major threat to public health worldwide. Recent research allowed breakthrough in our understanding of the virus and host pathogenesis; however, little is known on its impact on its main vector, Aedes aegypti. Here we show how Zika virus targets Aedes aegypti’s neurons and induces changes in its behavior. Results are compared to dengue virus, another flavivirus, which triggers a different pattern of behavioral changes. We used microelectrode array technology to record electrical spiking activity of mosquito primary neurons post infections and discovered that only Zika virus causes an increase in spiking activity of the neuronal network. Confocal microscopy also revealed an increase in synapse connections for Zika virus-infected neuronal networks. Interestingly, the results also showed that mosquito responds to infection by overexpressing glutamate regulatory genes while maintaining virus levels. This neuro-excitation, possibly via glutamate, could contribute to the observed behavioral changes in Zika virus-infected Aedes aegypti females. This study reveals the importance of virus-vector interaction in arbovirus neurotropism, in humans and vector. However, it appears that the consequences differ in the two hosts, with neuropathology in human host, while behavioral changes in the mosquito vector that may be advantageous to the virus.

Assessment of ICount software, a precise and fast egg counting tool for the mosquito vector Aedes aegypti

BackgroundWidespread in the tropics, the mosquito Aedes aegypti is an important vector of many viruses, posing a significant threat to human health. Vector monitoring often requires fecundity estimation by counting eggs laid by female mosquitoes. Traditionally, manual data analyses have been used but this requires a lot of effort and is the methods are prone to errors. An easy tool to assess the number of eggs laid would facilitate experimentation and vector control operations.ResultsThis study introduces a built-in software called ICount allowing automatic egg counting of the mosquito vector, Aedes aegypti. ICount egg estimation compared to manual counting is statistically equivalent, making the software effective for automatic and semi-automatic data analysis. This technique also allows rapid analysis compared to manual methods. Finally, the software has been used to assess p-cresol oviposition choices under laboratory conditions in order to test the system with different egg densities.ConclusionsICount is a powerful tool for fast and precise egg count analysis, freeing experimenters from manual data processing. Software access is free and its user-friendly interface allows easy use by non-experts. Its efficiency has been tested in our laboratory with oviposition dual choices of Aedes aegypti females. The next step will be the development of a mobile application, based on the ICount platform, for vector monitoring surveys in the field.

Activity and Flight Trajectory Monitoring of Mosquito Colonies for Automated Behaviour Analysis

Monitoring and tracking of mosquitoes using image processing is important to facilitate the mosquitos’ behaviour analysis automatically over longer period of times. In this paper, we propose a simple methodology to monitor mosquitos’ activity using multiple cameras optimally placed. In order to ensure optimal camera coverage for the area of observation and desired image quality; we propose to simulate the experimental setup in a 3D virtual environment to obtain one-off optimum camera placement parameters. Our proposed methodology is demonstrated to have improved the activity monitoring process using two cameras for accurate count of occluded mosquitoes and 3D trajectory path reconstruction. This framework will enable working out more challenging tasks of constructing 3D trajectories using information received from multiple low quality cameras, which provide inconsistent and discontinuous trajectories.

Visual effects in great bowerbird sexual displays and their implications for signal design (vol 281, 20140235, 2014)

We inadvertently left out one acknowledgement in our paper [1] and we add it now in this addendum: We thank Natalie Doerr for an excellent job of setting up and running the bowerbird cameras and other aspects of fieldwork during the 2012 season. In addition, we neglected to reduce the number of significant figures in table 1 from the original R output. If interested, please round off each number to three significant figures.

Zika virus-induced hyper excitation precedes death of mouse primary neuron

BackgroundZika virus infection in new born is linked to congenital syndromes, especially microcephaly. Studies have shown that these neuropathies are the result of significant death of neuronal progenitor cells in the central nervous system of the embryo, targeted by the virus. Although cell death via apoptosis is well acknowledged, little is known about possible pathogenic cellular mechanisms triggering cell death in neurons.MethodsWe used in vitro embryonic mouse primary neuron cultures to study possible upstream cellular mechanisms of cell death. Neuronal networks were grown on microelectrode array and electrical activity was recorded at different times post Zika virus infection. In addition to this method, we used confocal microscopy and Q-PCR techniques to observe morphological and molecular changes after infection.ResultsZika virus infection of mouse primary neurons triggers an early spiking excitation of neuron cultures, followed by dramatic loss of this activity. Using NMDA receptor antagonist, we show that this excitotoxicity mechanism, likely via glutamate, could also contribute to the observed nervous system defects in human embryos and could open new perspective regarding the causes of adult neuropathies.ConclusionsThis model of excitotoxicity, in the context of neurotropic virus infection, highlights the significance of neuronal activity recording with microelectrode array and possibility of more than one lethal mechanism after Zika virus infection in the nervous system.

Electrophysiological evidence of RML12 mosquito cell line towards neuronal differentiation by 20-hydroxyecdysdone

Continuous cell lines from insect larval tissues are widely used in different research domains, such as virology, insect immunity, gene expression, and bio pharmacology. Previous study showed that introduction of 20-hydroxyecdysone to Spodoptera cell line induced a neuron-like morphology with neurite extensions. Despite some results suggesting potential presence of neuro-receptors, no study so far has shown that these neuron-induced cells were functional. Here, using microelectrode arrays, we showed that the mosquito cell line, RML12, differentiated with 20-hydroxyecdysone, displays spontaneous electrophysiological activity. Results showed that these cells can be stimulated by GABAergic antagonist as well as nicotinic agonist. These results provide new evidence of neuron-like functionality of 20-hydroxyecdysone induced differentiated mosquito cell line. Finally, we used this new model to test the effects of two insecticides, temephos and permethrin. Our analysis revealed significant changes in the spiking activity after the introduction of these insecticides with prolonged effect on the neuronal activity. We believe that this differentiated mosquito neuronal cell model can be used for high-throughput screening of new pesticides on insect nervous system instead of primary neurons or in vivo studies.

Dengue virus infection changes Aedes aegypti oviposition olfactory preferences

Aedes aegypti mosquitoes, main vectors for numerous flaviviruses, have olfactory preferences and are capable of olfactory learning especially when seeking their required environmental conditions to lay their eggs. In this study, we showed that semiochemical conditions during Aedes aegypti larval rearing affected future female choice for oviposition: water-reared mosquitoes preferred to lay eggs in water or p-cresol containers, while skatole reared mosquitoes preferred skatole sites. Using two independent behavioural assays, we showed that this skatole preference was lost in mosquitoes infected with dengue virus. Viral RNA was extracted from infected female mosquito heads, and an increase of virus load was detected from 3 to 10 days post infection, indicating replication in the insect head and possibly in the central nervous system. Expression of selected genes, potentially implied in olfactory learning processes, were also altered during dengue infection. Based on these results, we hypothesise that dengue virus infection alters gene expression in the mosquito’s head and is associated with a loss of olfactory preferences, possibly modifying oviposition site choice of female mosquitoes.

Insects Neural Model: Potential Alternate to Mammals for Electrophysiological Studies

Microelectrode arrays are a promising tool in the study of electrophysiology of the brain in vitro. Allowing non-invasive recording of electrical signals from neuronal networks, the device is a perfect tool to help scientists to understand brain functioning at the cell level and as a whole system. Despite the large increase of MEA technology in terms of device diversity and use in neurophysiology, biological models in vitro are mostly restrained to mammals. This chapter highlights the advantages to combine insects and MEA technology for future neurophysiological studies, and introduce the possible perspectives of this research.