This thesis describes work from both the University of Cambridge in the lab of Berthold Hedwig and from the HHMI Janelia Research Campus in the lab of Michael Reiser. At the University of Cambridge, my work involved the development and demonstration of a method for electrophoretically delivering dyes and tracers for anatomical and functional imaging into animals that are not amenable to genetic labelling techniques. Using this method in locusts and crickets - model systems of particular interest for their acoustic communication - I successfully delivered polar fluorescent dyes and tracers through the sheath covering the auditory nerve, simultaneously staining both the peripheral sensory structures and the central axonal projections without destroying the nerve's function. I could label neurons which extend far from the tracer delivery site on the nerve as well as local neuron populations through the brain's surface. I used the same method to deliver calcium indicators into central neuropils for in vivo optical imaging of sound-evoked activity, as well as calling song-evoked activity in the brain. The work completed at the Janelia Research Campus began with the development of a modern version of a modular LED display and virtual reality control system to enable research on the visual control of complex behaviors in head-fixed animals. The primary advantages of our newly developed LED-based display over other display technologies are its high-speed operation, brightness uniformity and control, precise synchronization with analog inputs and outputs, and its ability to be configured into a variety of display geometries. Utilizing the system's fast display refresh rates, I conducted the first accurate characterization of the upper limits of the speed sensitivity of Drosophila for apparent motion during flight. I also developed a flexible approach to presenting optic flow scenes for functional imaging of motion-sensitive neurons. Finally, through the on-line analysis of behavioral measures, image rendering, and display streaming with low latency to multi-color (UV/Green) LED panels, I demonstrated the ability to create more naturalistic stimuli and interactive virtual visual landscapes. Lastly, I used this new visual display system to explore a newly discovered cell-type that had been implicated in higher-order motion processing from a large genetic screen of visually-guided behavior deficits. Using genetic silencing and activation methods, and by designing stimuli that modeled the optic flow encountered during different types of self-motion, colleagues in the Reiser lab and I showed that this cell-type - named Lobula Plate Columnar 1 (LPC1) - is required for the stopping behavior of walking flies caused by back-to-front translation motion but is not involved in the rotational optomotor response. Using calcium imaging, I found that LPC1 was selectively excited by back-to-front motion on the eye ipsilateral to the neuron population and inhibited by front-to-back motion on the contralateral eye, demonstrating a simple mechanism for its selectivity to translation over rotation. I also examined an anatomically similar cell type - named Lobula-Lobula Plate Columnar type 1 (LLPC1) - and found that its selectivity results from a similar but opposite calculation for the detection of front-to-back translational motion. The detection of back-to-front motion had previously been hypothesized to be useful for collision avoidance, and this work provides a neural mechanism for how this detection could be accomplished, as well as providing a platform from which to explore the larger network for translation optic flow.

Since several decades, field crickets of the subfamily Gryllinae under order Orthoptera have been used as a model group by researchers working in the field of neuroethology, behavioural ecology and bioacoustics. Subfamily Gryllinae is a much diversified group and members are diverse in their biological traits however the most commonly used model systems in the research fields belong to only three genera: Gryllus, Teleogryllus and Acheta. All the three genera are related to each other therefore, they fail to represent the wide diversity of ecological and behavioral traits that are encompassed by other members of Gryllinae. To understand the diversity in an evolutionary framework in field crickets, it is thus necessary to take into account the evolutionary background of the model systems. A phylogenetic analysis is a prerequisite for all evolutionary studies. A phylogeny hypothesizes the relationship among the studied taxa on the basis of homology drawn from all the characters that were used for the analysis. The phylogenetic tree is a branching diagram where related taxa are grouped together at nodes and the terminal ends of the branches are the studied taxa. The characters are mapped on to this tree thus indicating the hypothetical ancestral state of each character at the nodes and explaining the transformational stages of each character that gave rise to the current character state. This procedure helps in understanding the evolutionary history and understanding the patterns of relationships among taxa that in turn also help in identification of species. This process of identification and classification by phylogenetic reconstruction is formally known as “phylogenetic systematics”. The main goal of this thesis work was to understand the phylogenetic relationships among field crickets of the subfamily Gryllinae. This study is of importance in the field of orthopteran classification because since the 19th century, taxonomists proposed several taxonomic rankings and various classificatory schemes either by describing new classificatory levels or by upgrading existing taxonomic levels to higher levels. In addition, different sets of morphological characters have been used in proposing the classification hypotheses which in turn leads to the existence of several mutually exclusive hypotheses. Although there have been several changes in the taxonomic positioning of these crickets under Gryllinae, none so far have been based on a phylogenetic hypothesis. Therefore, I described 100 characters based on external and internal morphological structures of crickets belonging to 17 genera to test all the previously proposed classificatory hypotheses, provide a scheme for classifying these crickets under different hierarchical levels under Gryllinae and a strong base for conducting further evolutionary studies. The results lend full support to one of the previously proposed hypotheses for classification. Subfamily Gryllinae is a monophyletic clade where all the field crickets should be grouped under two tribes. The results also invalidate several tribes that have been erected in recent years. The second aim was to test species concepts by examining concordance in species boundaries generated from different data sets and to study the pattern of song evolution in field crickets. Therefore, for this study a field cricket belonging to genus Itaropsis was selected. Only one species of Itaropsis is known from the Indian subcontinent which was described from Sri Lanka. Interestingly, five call types of Itaropsis were encountered when a field survey was performed at four different localities in the Western Ghats of India. Males of the all five call types (varying from long trills to short chirps) looked morphologically very similar to each other. However, they differed in their calling song structure, I described four call features (carrier frqeuncy, syllable period, syllable duration and call duration) and classified the individuals into clusters by calculating distances using these call features. The clusters thus obtained were considered to reflect different species under the genus Itaropsis. To assure that the clusters thus obtained reflect different species, concordance between the results was tested by comparing these results with the clades of an available phylogeny for the collected individuals of this genus using both morphological and molecular data. The acoustic clusters showed concordance with the phylogenetic lineages largely however an absolute match was not found suggesting that these lineages cannot be called as distinct species. The pattern of evolution of songs in Itaropsis was examined by optimizing the call features on the phylogenetic tree derived from molecular data. This showed short chirps with a dominant frequency of 7 kHz to be the ancestral call type within this genus, from which long trills with long syllable durations have evolved. There were no significant patterns in the evolution of carrier frequency. In most crickets, the calling songs that are produced by males to attract their conspecific females are considered species specific therefore, the calling songs serve as an additional tool in identification of crickets. The third aim of my work was to use only call features of crickets in delineation of species but for higher number of species. Therefore, for this I used recorded calling songs of 14 species of field crickets and described their call features. I then used either five or seven call features to construct clusters based on Euclidean distances between the songs of all pairs of individuals. I also carried out a discriminant function analysis of these data to test the efficiency of species identification when taxa were specified a priori. I varied the number of species used for the analysis systematically from 5 to 14 to examine the effect of varying the number of taxa on correct species identification. The results showed that taxa number can be increased upto ten to obtain almost correct classification however, using six to seven taxa is optimal since they provide 90% accuracy in classification. To the best of my knowledge, this study on understanding the phylogenetic relationships among field crickets is the first to be carried out at subfamily level. Description of new call types reflecting multiple or incipient species under the genus Itaropsis and use of acoustic features in species identification of Indian field crickets is also a new contribution to the area of orthopteran systematics and phylogenetics.

Acoustic communication in orthopterans and anurans provides a suitable model system for studying the evolutionary mechanisms of sexual selection mainly because males use acoustic signals to attract females over long distances for pair formation. Females use these signals not only to localize conspecific males but also to discriminate between potential mates. Investigations on the effect of sexual selection on acoustic signals requires an understanding of how female preferences for different features of the acoustic signal affect male mating success under ecological constraints in wild populations. The effect of female preferences on male mating success depends on the mate sampling strategy that females employ to search for potential mates. Despite its relevance, female mate sampling strategies based on male acoustic signals have rarely investigated in orthopterans and anurans, especially in the field. Considering the elaborate knowledge of the role of sensory physiology in female phonotaxis behaviour and characterization of the male acoustic signal, I used the field cricket species Plebeiogryllus guttiventris as a model system in this study. In this thesis, I first investigated the ecology of callers in wild populations. I then investigated female mate sampling strategies by incorporating relevant information on the ecology of signalers and the sensory physiology of receivers. Amount of calling activity is a strong determinant of male mating success in acoustically communicating species such as orthopterans and anurans. While many studies in crickets have investigated the determinants of calling effort, patterns of variability in male calling effort in natural choruses remain largely unexplored. I therefore investigated the spatio-temporal dynamics of acoustic chorusing behaviour in a wild population. I first studied the consistency of calling activity by quantifying variation in male calling effort across multiple nights of calling using repeatability analysis. Callers were inconsistent in their calling effort across nights and did not optimize nightly calling effort to increase their total number of nights spent calling. Next, I investigated calling site fidelity of males across multiple nights by quantifying movement of callers. Callers frequently changed their calling site across calling nights with substantial displacement but without any significant directionality. Finally, I investigated trade-offs between within-night calling effort and energetically expensive calling song features such as call intensity and chirp rate. Calling effort was not correlated with any of the calling song features, suggesting that energetically expensive song features do not constrain male calling effort. The two key features of signaling behaviour, calling effort and call intensity, which determine the duration and spatial coverage of the sexual signal, are uncorrelated and function independently Acoustic signal variation and female preference for different signal components constitute the prerequisite framework to study the mechanisms of sexual selection that shape acoustic communication. Despite several studies of acoustic communication in crickets, information on both male calling song variation in the field and female preference in the same system is lacking for most species. First, I quantified variation in the spectral, temporal and amplitudinal characteristics of the male calling song in a wild population, at two temporal scales, within and across nights, using repeatability analysis. Carrier frequency (CF) was the most repeatable call trait across nights, whereas chirp period (CP) had low repeatability. I further investigated female preferences based on song features with high and low repeatability (CF and CP respectively). Females showed no consistent preferences for CF but were more attracted towards calls with higher rates (shorter CP). I also examined the effect of signal intensity, which is known to play a critical role in female phonotaxis behaviour, on female preferences for faster calls. Females preferred louder calls over faster ones, implying a dominant role for signal intensity in female evaluation of potential mates based on acoustic signals. Call intensity was also the only signal feature that was positively correlated with male size. In the final chapter, I investigated female mate sampling strategies based on acoustic signals using both theoretical and empirical approaches. Analytical models of mate sampling have demonstrated significant differences in individual fitness returns for different sampling strategies. However these models have rarely incorporated relevant information on the ecology of signalers and the sensory physiology of receivers. I used simulation models to compare the costs and benefits of different mate sampling strategies by incorporating information on relative spacing of callers in natural choruses and the effect of signal intensity on female phonotaxis behaviour. The strategy of mating with males that were louder at the female position emerged as the optimal sampling rule in the simulations. When tested empirically in the field using callers in natural choruses, females seemed to follow the optimal strategy of mating with males that were perceived as louder at their position.

Chez les espèces sociales, les vocalisations transmettent des informations qui participent au maintien et à la survie du groupe. Alors que de nombreuses études se sont intéressées aux informations stables portées par les vocalisations telles que l’identité, peu d’études se sont interrogées sur le rôle des signaux vocaux dans la transmission des informations plus labiles, telles que l’état émotionnel de l’émetteur. Le stress est un bon candidat pour l’étude de l’expression des émotions chez les animaux, puisqu’il est directement mesurable via un dosage de la concentration plasmatique en glucocorticoïdes. Le stress est connu pour modifier les paramètres acoustiques des vocalisations chez les mammifères, mais peu d’études ont traité ce processus chez les oiseaux, qui présentent pourtant des réseaux sociaux complexes. Le but de cette thèse est de déterminer de quelle manière les oiseaux expriment vocalement leur stress, et d’évaluer dans quelle mesure la corticostérone, hormone de stress principale chez l’oiseau, est impliquée dans le phénomène. Je me suis intéressée au Diamant mandarin (Taenopygia guttata), un oiseau chanteur australien au comportement grégaire qui forme des liens d’appariements à vie et prodigue des soins biparentaux à sa progéniture. En administrant de la corticostérone exogène à des oiseaux et en utilisant également des évènements sociaux stressants, nous montrons que les mâles adultes et les poussins expriment leur stress à travers l’émission de cris modifiés dans leur structure, ce qui suggère que la flexibilité des cris chez les oiseaux est plus importante que l’avaient montré des études précédentes. Par une analyse complète des paramètres temporels et spectraux des cris, nous montrons pour la première fois que le stress, par un effet direct de la corticostérone, déclenche l’émission de vocalisations présentant un spectre de fréquence déplacé vers les hautes fréquences. De plus, les receveurs du signal (respectivement les partenaires femelles et les parents) semblent capables de décoder l’information portée par ces cris de stress car ils modifient leur comportement en conséquence. Les processus physiques impliqués dans l’émission de cris modulés par le stress sont également discutés en appliquant la théorie « source-filtre » généralement utilisée chez les mammifères. Enfin, les valeurs adaptatives de ces cris sont également envisagées, en rapport avec le réseau social du Diamant mandarin et les risques de prédation encourus par l’émetteur du signal. Ce travail apporte de nouvelles preuves sur l’expression du stress chez les oiseaux, et propose une étude complète, des signaux physiologiques impliqués dans le stress aux modifications de comportement de l’émetteur, qui déclenchent une réponse adaptative des receveurs du signal / In social species, vocalisations convey information that participates in the maintenance and the survival of the group. While many studies were interested in stable information carried by vocal signals, like identity, fewer studies dealt with their potential role in informing about labile information such as the senders’ emotional state. Stress is a good candidate for the study of the expression of emotions in animals, as it is directly measurable by the plasma levels of glucocorticoïds. Stress is known to modify acoustic parameters of vocalisations in mammals, but few studies studied the process in birds, that also show complex social networks. The aim of this thesis is thus to determine how birds can vocally express their stress and to what extent corticosterone, the main stress hormone in birds, is implicated in this expression. I focused my research on the zebra finch (Taenopygia guttata), a gregarious Australian songbird that form lifelong pairbond and provides biparental care to its young. Using oral administration of exogenous corticosterone but also social stressful events, we show that both adult males and nestlings can express their stress through modulations of their calls’ structure, suggesting that flexibility in birds’ vocalisations is higher than previously expected. With a complete analysis of temporal and spectral parameters of calls, we show for the first time that stress evokes the emission of vocalisations with up-shifted frequency spectrum via a direct effect of corticosterone. Moreover, we show that females and parents are able to decode the information carried by stressed-induced calls of respectively their male partner and their young, as they exhibit modifications of behaviour in accordance with the context. The physical processes leading to the emission of stressed-induced vocalisations are discussed by applying the source-filter theory usually used in mammals. Adaptive values are also proposed, in regards with the zebra finch social network and predation risks for the caller. This work gives new evidences about the expression of stress in birds, and proposes a comprehensive study, from the physiological signals involved in stress to the resulting modifications of communication behaviour for the sender, that leads to an adaptive response from the receivers

Pour comprendre comment la sélection naturelle modèle les traits sexuels, l’emploi d’une approche intégrative des processus sélectifs en jeu semble aujourd’hui indispensable. Durant ma thèse je me suis ainsi intéressé aux influences simultanées des sélections de viabilité et sexuelle sur l’origine, l’évolution et le maintien des traits sexuels de la petite teigne de la ruche (Achroia grisella). Chez cette pyrale les mâles se regroupent en leks, produisent un chant d’appel ultrasonique intense et les femelles choisissent un partenaire sexuel en se basant sur des caractéristiques acoustiques spécifiques de ce chant. Cependant les signaux sexuels ultrasoniques des mâles peuvent également attirer des prédateurs : les chauves-souris insectivores. Ainsi, au sein des leks, les sélections sexuelle et de viabilité jouent et peuvent modeler (1) l’évolution du choix des femelles, (2) le maintien et l’évolution du lek, (3) l’origine de la communication acoustique sexuelle. Les travaux conduits durant cette thèse soulignent l’intérêt de l’étude du jeu des sélections sexuelle et de viabilité pour comprendre comment la sélection naturelle peut modeler l’origine, l’évolution et le maintien des traits sexuels. Les résultats indiquent en particulier le rôle crucial de la pression de prédation dans ces processus sélectifs. / In order to achieve a better understanding of how natural selection shapes sexual traits it is essential to consider the interplay of the various selective processes at work. For my thesis I investigated the simultaneous influences of viability and sexual selections on the origin, the evolution and the maintenance of sexual traits in the lesser waxmoth Achroia grisella. Females of this pyralid moth choose their mate based on acoustic characters of the ultrasonic calling song broadcasted by males while competing at lek. However male ultrasonic displays are very conspicuous and may attract specific predators: insectivorous bats. Therefore at leks sexual and viability selection pressures act simultaneously and can shape (1) the evolution of female mate choice, (2) the maintenance and evolution of lekking, (3) the origin of sexual acoustic communication. Taken as a whole this work emphasises the relevance of the study of selective interplays between viability and sexual selections in order to improve our understanding of how natural selection may shape the origin, evolution and maintenance of sexual traits. In particular results indicate the crucial role played by predation pressure within these evolutionary processes.

[pt] A necessidade de transmitir energia e dados através de barreiras metálicas tem sido cada vez maior em aplicações industriais, onde não é possível a penetração de cabos elétricos, ou o uso de ondas eletromagnéticas, tais como, por exemplo, em sistemas de sensoriamento de cimentação em poços de petróleo. Ondas acústicas podem ser uma solução para esse problema, porque não são afetadas pelo efeito gaiola de Faraday, além de possuírem baixa atenuação ao atravessarem metais. Diversos esforços foram feitos para realizar a transmissão de dados através de camadas metálicas, com abordagens que variam em composição do canal acústico, taxa de transmissão, transmissão simultânea de dados e energia e complexidade dos circuitos empregados; existe, porém, carência de trabalhos que envolvam camadas metal-fluído-metal. Este trabalho apresenta uma possível solução utilizando ondas acústicas como meio de transportar energia e dados em um canal composto de barreiras com duas camadas metálicas e uma de fluído. Aqui propõe-se uma inovadora técnica de controle automático de ganho e um melhor aproveitamento da largura de banda do canal acústico, que permite maior taxa de transmissão de dados. É ainda proposta uma técnica para controle dinâmico da portadora enviada ao lado passivo do sistema. Inicialmente, foi feita uma análise de um modelo numérico, baseado em trabalhos anteriores, fundamentado na propagação de ondas acústicas e baseado na analogia acustoelétrica. Em seguida, desenvolveu-se um sistema eletrônico para receber / transmitir energia e dados digitais, modulados em frequência, de um lado ao outro do sistema. Por fim, análises experimentais foram feitas utilizando como canal acústico, um conjunto de duas placas planas de aço (de 5 mm) separadas por uma camada de fluído (de 100 mm) e dois transdutores alinhados axialmente, realizando a transferência de energia e dados digitais modulados em frequência. O sistema foi capaz de realizar a transferência de dados a uma taxa de 19200 bps e simultaneamente uma transferência de energia de 66 mW, com essa energia foi possível alimentar o modulo eletrônico e um sensor de pressão e temperatura. Durante os testes foi constatado um aproveitamento de 5,5 por cento da energia aplicada ao canal, e foi possível atingir uma taxa de erro de bit de 5 por cento em um teste com 2 h e 30 min de duração, utilizando o canal acústico com camadas de múltiplos materiais propostos. O sistema de controle de portadora funcionou adequadamente, permitindo uma redução de consumo de até 53 por cento. O controle automático de ganho permitiu uma redução de 50 por cento na taxa de erro de decodificação. Demonstra-se, então, a viabilidade de tais sistemas de controle propostos, os quais podem ser úteis em casos onde existam variações nas características acústicas do canal em questão que, em conjunto com a transferência não intrusiva, pode prover solução para sistemas de sensoriamento. / [en] The need for energy and data transmission through metallic barriers is increasing in industrial applications, where the penetration of electrical waves or the use of electrical waves is not possible. An example of such a scenario is the monitoring of cementing in wellbore applications. Acoustic waves are promising to solve this problem, since they are not affected by the Faraday cage effect, in addition, they present low attenuation when propagating in metals. Several efforts have been made to carry out data transmission through metallic layers, with approaches that vary in composition of the acoustic channel, transmission speed, simultaneous transmission of data and energy and the complexity of the circuits used, but there is a lack of works involving metal-fluid-metal layers. This work presents a possible solution using acoustic waves as a mean of transporting energy and data in a channel composed of barriers with two metallic layers and one fluid layer. Here, it is proposed a novel technique for automatic gain control and better use of the available bandwidth of the acoustic channel, which allows higher data transmission speed. Also, a technique for dynamic control of the carrier sent to the passive side of the system is proposed. Initially, an analysis with a numerical model was made, following previous works, which is based on the propagation of acoustic waves and relying on the acoustoelectric analogy. Then, an electronic system was developed to receive/transmit power and digital data, frequency modulated, from one side of the system to the other. Finally, experimental analyzes were performed using as an acoustic channel, a set of two flat steel plates (5 mm) separated by a fluid layer (100 mm) and a pairs of axially aligned transducers, performing the energy transfer and frequency modulated digital data. The system was able to transfer data at a rate of 19200 bps and simultaneously a transfer of energy of 66 mW, with this energy it was possible to feed the inside block module and a pressure and temperature sensor. During the tests, it was verified that 5.5 per cent of the energy applied to the channel was used, and it a bit error rate down to of 5 per cent was reached in a test with 2 h and 30 min of duration, using the multi-layered acoustic. The automatic carrier control system worked as expected and allowed one to reduce energy consumption in 53 per cent. The automatic gain control allowed one to reduce the error rate in 50 per cent. These control systems prove the feasibility of the proposed system and further show the usefulness of the system in scenarios that are subject to variations in the acoustic characteristics of the channel.