Electric Organ Discharge Research Articles

Ontogeny of electric organ and electric organ discharge in Campylomormyrus rhynchophorus (Teleostei: Mormyridae)

The aim of this study was a longitudinal description of the ontogeny of the adult electric organ of Campylomormyrus rhynchophorus which produces as adult an electric organ discharge of very long duration (ca. 25 ms). We could indeed show (for the first time in a mormyrid fish) that the electric organ discharge which is first produced early during ontogeny in 33-mm-long juveniles is much shorter in duration and has a different shape than the electric organ discharge in 15-cm-long adults. The change from this juvenile electric organ discharges into the adult electric organ discharge takes at least a year. The increase in electric organ discharge duration could be causally linked to the development of surface evaginations, papillae, at the rostral face of the electrocyte which are recognizable for the first time in 65-mm-long juveniles and are most prominent at the periphery of the electrocyte.

The transcriptional correlates of divergent electric organ discharges in Paramormyrops electric fish

BackgroundUnderstanding the genomic basis of phenotypic diversity can be greatly facilitated by examining adaptive radiations with hypervariable traits. In this study, we focus on a rapidly diverged species group of mormyrid electric fish in the genus Paramormyrops, which are characterized by extensive phenotypic variation in electric organ discharges (EODs). The main components of EOD diversity are waveform duration, complexity and polarity. Using an RNA-sequencing based approach, we sought to identify gene expression correlates for each of these EOD waveform features by comparing 11 specimens of Paramormyrops that exhibit variation in these features.ResultsPatterns of gene expression among Paramormyrops are highly correlated, and 3274 genes (16%) were differentially expressed. Using our most restrictive criteria, we detected 145–183 differentially expressed genes correlated with each EOD feature, with little overlap between them. The predicted functions of several of these genes are related to extracellular matrix, cation homeostasis, lipid metabolism, and cytoskeletal and sarcomeric proteins. These genes are of significant interest given the known morphological differences between electric organs that underlie differences in the EOD waveform features studied.ConclusionsIn this study, we identified plausible candidate genes that may contribute to phenotypic differences in EOD waveforms among a rapidly diverged group of mormyrid electric fish. These genes may be important targets of selection in the evolution of species-specific differences in mate-recognition signals.

Effects of hypoxia on aerobic metabolism and active electrosensory acquisition in the African weakly electric fish Marcusenius victoriae.

Environmental hypoxia has effected numerous and well-documented anatomical, physiological and behavioural adaptations in fishes. Comparatively little is known about hypoxia's impacts on sensing because it is difficult to quantify sensory acquisition in vivo. Weakly electric fishes, however, rely heavily on an easily-measurable sensory modality-active electric sensing-whereby individuals emit and detect electric organ discharges (EODs). In this study, hypoxia tolerance of a mormyrid weakly electric fish, Marcusenius victoriae, was assessed by examining both its metabolic and EOD rates using a critical threshold (pcrit ) paradigm. The routine metabolic rate was 1.42 mg O2 h-1 , and the associated critical oxygen tension was 14.34 mmHg. Routine EOD rate was 5.68 Hz with an associated critical tension of 15.14 mmHg. These metabolic indicators of hypoxia tolerance measured in this study were consistent with those in previous studies on M. victoriae and other weakly electric fishes. Furthermore, our results suggest that some aerobic processes may be reduced in favour of maintaining the EOD rate under extreme hypoxia. These findings underscore the importance of the active electrosensory modality to these hypoxia-tolerant fish.

Electrocommunication in pulse Gymnotiformes: the role of electric organ discharge (EOD) time course in species identification.

Understanding how individuals detect and recognize signals emitted by conspecifics is fundamental to discussions of animal communication. The species pair Gymnotus omarorum and Brachyhypopomus gauderio, found in syntopy in Uruguay, emit species-specific electric organ discharge (EOD) that can be sensed by both species. The aim of this study was to unveil whether either of these species is able to identify a conspecific EOD, and to investigate distinctive recognition signal features. We designed a forced-choice experiment using a natural behavior (i.e. tracking electric field lines towards their source) in which each fish had to choose between a conspecific and a heterospecific electric field. We found a clear pattern of preference for a conspecific waveform even when pulses were played within 1 Hz of the same rate. By manipulating the time course of the explored signals, we found that the signal features for preference between conspecific and heterospecific waveforms were embedded in the time course of the signals. This study provides evidence that pulse Gymnotiformes can recognize a conspecific exclusively through species-specific electrosensory signals. It also suggests that the key signal features for species differentiation are probably encoded by burst coder electroreceptors. Given these results, and because receptors are sharply tuned to amplitude spectra and also tuned to phase spectra, we extend the electric color hypothesis used in the evaluation of objects to apply to communication signals.

Tethered unitary recordings suggest a spike-timing electrosensory code in the electrosensory lobe of Gymnotus omarorum

AbstractEvaluation of neural activity during natural behaviours is essential for understanding how the brain works. Here we show that neuron-specific self-evoked firing patterns are modulated by an object’s presence, at the electrosensory lobe neurons of tethered-moving Gymnotus omarorum. This novel preparation shows that electrosensory signals in these pulse-type weakly electric fish are not only encoded in the number of spikes per electric organ discharge (EOD), as is the case in wave-type electric fish, but also in the spike timing pattern after each EOD, as found in pulse-type Mormyroidea. Present data suggest that pulsant electrogenesis and spike timing coding of electrosensory signals developed concomitantly in the same species, and evolved convergently in African and American electric fish.

Tracking activity patterns of a multispecies community of gymnotiform weakly electric fish in their neotropical habitat without tagging.

Field studies on freely behaving animals commonly require tagging and often are focused on single species. Weakly electric fish generate a species- and individual-specific electric organ discharge (EOD) and therefore provide a unique opportunity for individual tracking without tagging. Here, we present and test tracking algorithms based on recordings with submerged electrode arrays. Harmonic structures extracted from power spectra provide fish identity. Localization of fish based on weighted averages of their EOD amplitudes is found to be more robust than fitting a dipole model. We apply these techniques to monitor a community of three species, Apteronotus rostratus, Eigenmannia humboldtii and Sternopygus dariensis, in their natural habitat in Darién, Panama. We found consistent upstream movements after sunset followed by downstream movements in the second half of the night. Extrapolations of these movements and estimates of fish density obtained from additional transect data suggest that some fish cover at least several hundreds of meters of the stream per night. Most fish, including E. humboldtii, were traversing the electrode array solitarily. From in situ measurements of the decay of the EOD amplitude with distance of individual animals, we estimated that fish can detect conspecifics at distances of up to 2 m. Our recordings also emphasize the complexity of natural electrosensory scenes resulting from the interactions of the EODs of different species. Electrode arrays thus provide an unprecedented window into the so-far hidden nocturnal activities of multispecies communities of weakly electric fish at an unmatched level of detail.

Development of a sexual dimorphism in a central pattern generator driving a rhythmic behavior: The role of glia-mediated potassium buffering in the pacemaker nucleus of the weakly electric fish Apteronotus leptorhynchus.

Central pattern generators play a critical role in the neural control of rhythmic behaviors. One of their characteristic features is the ability to modulate the oscillatory output. An important yet little-studied type of modulation involves the generation of oscillations that are sexually dimorphic in frequency. In the weakly electric fish Apteronotus leptorhynchus, the pacemaker nucleus serves as a central pattern generator that drives the electric organ discharge of the fish in a one-to-one fashion. Males discharge at higher frequencies than females-a sexual dimorphism that develops under the influence of steroid hormones. The two principal neurons that constitute the oscillatory network of the pacemaker nucleus are the pacemaker and relay cells. Whereas the number and size of the pacemaker and relay cells are sexually monomorphic, pronounced sex-dependent differences exist in the morphology, and subcellular properties of astrocytes, which form a syncytium closely associated with these neurons. In females, compared to males, the astrocytic syncytium covers a larger area surrounding the pacemaker and relay cells and exhibits higher levels of expression of connexin-43 expression. The latter indicates a strong gap-junction coupling of the individual cells within the syncytium. It is hypothesized that these sex-specific differences result in an increased capacity for buffering of extracellular potassium ions, thereby lowering the potassium equilibrium potential, which, in turn, leads to a decrease in the oscillation frequency. This hypothesis has received strong support from simulations based on computational models of individual neurons and the whole neural network of the pacemaker nucleus.

Silencing the Spark: CRISPR/Cas9 Genome Editing in Weakly Electric Fish.

Electroreception and electrogenesis have changed in the evolutionary history of vertebrates. There is a striking degree of convergence in these independently derived phenotypes, which share a common genetic architecture. This is perhaps best exemplified by the numerous convergent features of gymnotiforms and mormyrids, two species-rich teleost clades that produce and detect weak electric fields and are called weakly electric fish. In the 50 years since the discovery that weakly electric fish use electricity to sense their surroundings and communicate, a growing community of scientists has gained tremendous insights into evolution of development, systems and circuits neuroscience, cellular physiology, ecology, evolutionary biology, and behavior. More recently, there has been a proliferation of genomic resources for electric fish. Use of these resources has already facilitated important insights with regards to the connection between genotype and phenotype in these species. A major obstacle to integrating genomics data with phenotypic data of weakly electric fish is a present lack of functional genomics tools. We report here a full protocol for performing CRISPR/Cas9 mutagenesis that utilizes endogenous DNA repair mechanisms in weakly electric fish. We demonstrate that this protocol is equally effective in both the mormyrid species Brienomyrus brachyistius and the gymnotiform Brachyhypopomus gauderio by using CRISPR/Cas9 to target indels and point mutations in the first exon of the sodium channel gene scn4aa. Using this protocol, embryos from both species were obtained and genotyped to confirm that the predicted mutations in the first exon of the sodium channel scn4aa were present. The knock-out success phenotype was confirmed with recordings showing reduced electric organ discharge amplitudes when compared to uninjected size-matched controls.

The effect of normoxia exposure on hypoxia tolerance and sensory sampling in a swamp-dwelling mormyrid fish

Effects of energetic limitations on the performance of sensory systems are generally difficult to quantify. Weakly electric fishes provide an ideal model system to quantify the effects of metabolic stressors on sensory information acquisition, because they use an active-sensing strategy that permits easy measurement of the sensing effort. These fishes discharge an electric signal and sense perturbations of the resulting electric field. We used the mormyrid Petrocephalus degeni to quantify the relationship between routine metabolic rate and the rate of sensory sampling (rate of electric organ discharge, EOD) while under progressive hypoxia by quantifying the critical oxygen tension (PC-MR) and the critical electric organ discharge threshold (PC-EOD). PC-MR was significantly higher in fish acclimated to normoxia for over 40 days compared to animals tested within 1–5 days of capture from a hypoxic swamp, which suggests high costs of maintaining hypoxia tolerance; however, there was no acclimation effect on PC-EOD. All P. degeni reached their PC-EOD prior to their PC-MR. However, below the respective critical tension value, EOD rate decreased more gradually than the metabolic rate suggesting that the fish were increasing the proportion of their energy budget allocated to acquiring sensory information as dissolved-oxygen levels dropped. Trade-offs between sensory sampling and other physiological functions are also suggested by the increase in routine EOD rate with long-term normoxia acclimation, in contrast to metabolic rate, which showed no significant changes. These results highlight the relationship between sensory sampling and metabolic rate in response to progressive hypoxia and the plasticity of hypoxia tolerance.

A JAR of Chirps: The Gymnotiform Chirp Can Function as Both a Communication Signal and a Jamming Avoidance Response.

The weakly electric gymnotiform fish produce a rhythmic electric organ discharge (EOD) used for communication and active electrolocation. The EOD frequency is entrained to a medullary pacemaker nucleus. During communication and exploration, this rate can be modulated by a pre-pacemaker network, resulting in specific patterns of rate modulation, including stereotyped communication signals and dynamic interactions with conspecifics known as a Jamming Avoidance Response (JAR). One well-known stereotyped signal is the chirp, a brief upward frequency sweep usually lasting less than 500 ms. The abrupt change in frequency has dramatic effects on phase precession between two signalers. We report here on chirping in Brachyhypopmus cf. sullivani, Microsternarchus cf. bilineatus Lineage C, and Steatogenys cf. elegans during conspecific playback experiments. Microsternarchus also exhibits two behaviors that include chirp-like extreme frequency modulations, EOD interruptions with hushing silence and tumultuous rises, and these are described in terms of receiver impact. These behaviors all have substantial impact on interference caused by conspecifics and may be a component of the JAR in some species. Chirps are widely used in electronic communications systems, sonar, and other man-made active sensing systems. The brevity of the chirp, and the phase disruption it causes, makes chirps effective as attention-grabbing or readiness signals. This conforms to the varied assigned functions across gymnotiforms, including pre-combat aggressive or submissive signals or during courtship and mating. The specific behavioral contexts of chirp expression vary across species, but the physical structure of the chirp makes it extremely salient to conspecifics. Chirps may be expected in a wide range of behavioral contexts where their function depends on being noticeable and salient. Further, in pulse gymnotiforms, the chirp is well structured to comprise a robust jamming signal to a conspecific receiver if specifically timed to the receiver’s EOD cycle. Microsternarchus and Steatogenys exploit this feature and include chirps in dynamic jamming avoidance behaviors. This may be an evolutionary re-use of a circuitry for a specific signal in another context.

An Aquatic Wireless Biosensor for Electric Organ Discharge With an Integrated Analog Front End

This paper presents a novel wireless underwater data acquisition system for sensing electric organ discharge (EOD) signals generated from the weakly electric fish S. macrurus . Variation in frequency and amplitude of the EOD signals are of interest by biologists to study behavioral and environmental effects on electric organ cells. In order to record the EOD signals, a miniature wearable wireless sensing system is designed for the target fish. The system consists of a custom-designed integrated analog front-end (AFE), an ATmega328p microcontroller unit and a wireless transmitter (TX), as well as a battery with a power management module. In order to save TX power, the wireless sensor only transmits the calculated frequency and amplitude information of the EOD signal based on a proposed gain-feedback control method. A receiver attached to a host computer receives the wirelessly transmitted EOD data to perform further analysis. A wearable waterproof backpack for the fish is designed to house the wireless sensor and battery. The overall system has been successfully tested in a clinical experiment with the weakly electric fish. The AFE integrated circuit, which includes a novel rail-to-rail dynamic comparator, is fabricated in a 0.18- $\mu \text{m}$ CMOS process. The total area of the core circuit is 1.0 mm2. The LNPA achieved a noise efficiency factor of 1.8. The AFE power consumption is 2.2 $\mu \text{W}$ from a single +1.8 V supply, whereas the overall system power consumption is 15 mW. The proposed gain-feedback control structure for frequency and amplitude measurement saves the transmitter data rate by 3,756 times compared with sending the raw data.

Predation and Crypsis in the Evolution of Electric Signaling in Weakly Electric Fishes

Eavesdropping by electroreceptive predators poses a conflict for weakly electric fish, which depend on their Electric Organ Discharge (EOD) signals both for navigation and communication in the dark. The EODs that allow weakly electric fish to electrolocate and communicate in the dark may attract electroreceptive predators such as catfishes and Electric Eels. These predators share with their prey the synapomorphy of passive electric sense supported by ampullary electroreceptors that are highly sensitive to low-frequency electric fields. Any low-frequency spectral components of the EOD make weakly electric fish conspicuous and vulnerable to attack from electroreceptive predators. Accordingly, most weakly electric fish shift spectral energy upwards or cloak low-frequency energy with compensatory masking signals. Subadults and females in particular emit virtually no low-frequency energy in their EODs, whereas courting males include a significant low-frequency component, which likely attracts females, but makes the signals conspicuous to predators. Males of species that coexist with the most predators tend to produce the least low-frequency signal energy, expressing sexual dimorphism in their signals in less risky ways. In these respects, electric signals follow the classic responses to opposing forces of natural and sexual selection, as exemplified in the visual signals of guppies and the acoustic signals of Tungara frogs. Unique to electric fish is that the electric signal modifications that help elude detection by electroreceptive predators are additions to the basal signal rather than losses of attractive components. These enhancements that enable crypsis are energetically costly, but have also provided the evolutionary substrates for subsequent sexual selection and species identity characters.

Dominance in Habitat Preference and Diurnal Explorative Behavior of the Weakly Electric Fish Apteronotus leptorhynchus.

Electrocommunication and -localization behaviors of weakly electric fish have been studied extensively in the lab, mostly by means of short-term observations on constrained fish. Far less is known about their behaviors in more natural-like settings, where fish are less constrained in space and time. We tracked individual fish in a population of fourteen brown ghost knifefish (Apteronotus leptorhynchus) housed in a large 2 m3 indoor tank based on their electric organ discharges (EOD). The tank contained four different natural-like microhabitats (gravel, plants, isolated stones, stacked stones). In particular during the day individual fish showed preferences for specific habitats which provided appropriate shelter. Male fish with higher EOD frequencies spent more time in their preferred habitat during the day, moved more often between habitats during the night, and less often during the day in comparison to low-frequency males. Our data thus revealed a link between dominance indicated by higher EOD frequency, territoriality, and a more explorative personality in male A. leptorhynchus. In females, movement activity during both day and night correlated positively with EOD frequency. In the night, fish of either sex moved to another habitat after about 6 s on average. During the day, the average transition time was also very short at about 20 s. However, these activity phases were interrupted by phases of inactivity that lasted on average about 20 min during the day, but only 3 min in the night. The individual preference for daytime retreat sites did not reflect the frequent explorative movements at night.

Sexual Size Dimorphism in the Macana Tigrina, Gymnotus javari (Gymnotidae, Gymnotiformes)

Gymnotiform electric fishes exhibit many forms of sexual dimorphism, including sexual size dimorphism (SSD), aspects of head, jaw, and electric organ morphology, and electric signal waveform. However, sexual dimorphism is poorly documented in banded electric fishes (Gymnotus), where most species are monomorphic in external appearance and electric signal waveforms. Gathering comparative data on species of Gymnotus has historically been hampered by their predatory and territorial lifestyles, in that they are usually dispersed in their preferred habitats and often collected individually. Here data are presented from a population of G. javari from the Peruvian Amazon, examined for SSD (n = 55) and electric organ discharge (EOD) amplitude (n = 162). These results are compared with a syntopic population of G. carapo (n = 61), and also with G. omarorum (n = 24) from a population in Uruguay. Unlike most congeners, G. javari exhibits pronounced sexual dimorphism in total length and body weight, and EOD pulse amplitude is correlated with total length. A review of several ecological hypotheses on body-size distributions of each sex as a balance of multiple competing selection pressures finds none of these hypotheses adequately explain SSD in G. javari. Examining SSD in biogeographic and phylogenetic contexts leads to the conclusion that selection across different environments (e.g., local habitats, geographic regions) and through evolutionary time may have contributed to the evolution of SSD in some but not all lineages of Gymnotus.

Neural activity in a hippocampus-like region of the teleost pallium is associated with active sensing and navigation.

Most vertebrates use active sensing strategies for perception, cognition and control of motor activity. These strategies include directed body/sensor movements or increases in discrete sensory sampling events. The weakly electric fish, Gymnotus sp., uses its active electric sense during navigation in the dark. Electric organ discharge rate undergoes transient increases during navigation to increase electrosensory sampling. Gymnotus also use stereotyped backward swimming as an important form of active sensing that brings objects toward the electroreceptor dense fovea-like head region. We wirelessly recorded neural activity from the pallium of freely swimming Gymnotus. Spiking activity was sparse and occurred only during swimming. Notably, most units tended to fire during backward swims and their activity was on average coupled to increases in sensory sampling. Our results provide the first characterization of neural activity in a hippocampal (CA3)-like region of a teleost fish brain and connects it to active sensing of spatial environmental features.

Glia-mediated modulation of extracellular potassium concentration determines the sexually dimorphic output frequency of a model brainstem oscillator

Sexual dimorphism in behavior is widespread among animals, but the cellular mechanisms underlying neural control of this phenomenon are largely unknown. One behavior that has provided some important clues about how such sex differences might develop is the electric organ discharge of Apteronotus leptorhynchus. In this weakly electric fish, the mean discharge frequencies of males and females are 880 Hz and 740 Hz, respectively, with little overlap of the two frequency bands. The discharges are controlled, in a one-to-one fashion, by the neural oscillations of the pacemaker nucleus in the medulla oblongata. Experimental evidence has shown that the astrocytic syncytium associated with the neural network that generates these oscillations is significantly larger, and stronger coupled via gap junctions, in females than in males. In the present study, modeling of this network was performed to test the hypotheses that the sex-dependent differences in the structure and properties of the astrocytic syncytium mediate better buffering of extracellular potassium in females than in males, which in turn causes, via a lowering of the potassium equilibrium potential, a decrease in the oscillation frequency. Simulations of the neural activity of the pacemaker nucleus and its individual components demonstrated that under both spontaneous and induced conditions the oscillation frequency and the potassium equilibrium potential are strongly positively correlated. These simulations predict that sufficient separation of the electric organ discharge frequencies for establishment of the sexual dimorphism can be achieved by rather minor alterations in the concentration of the extracellular potassium concentration in the pacemaker nucleus.

Electroreception, electrogenesis and electric signal evolution.

Electroreception, the capacity to detect external underwater electric fields with specialised receptors, is a phylogenetically widespread sensory modality in fishes and amphibians. In passive electroreception, a capacity possessed by c. 16% of fish species, an animal uses low-frequency-tuned ampullary electroreceptors to detect microvolt-range bioelectric fields from prey, without the need to generate its own electric field. In active electroreception (electrolocation), which occurs only in the teleost lineages Mormyroidea and Gymnotiformes, an animal senses its surroundings by generating a weak (< 1 V) electric-organ discharge (EOD) and detecting distortions in the EOD-associated field using high-frequency-tuned tuberous electroreceptors. Tuberous electroreceptors also detect the EODs of neighbouring fishes, facilitating electrocommunication. Several other groups of elasmobranchs and teleosts generate weak (< 10 V) or strong (> 50 V) EODs that facilitate communication or predation, but not electrolocation. Approximately 1.5% of fish species possess electric organs. This review has two aims. First, to synthesise our knowledge of the functional biology and phylogenetic distribution of electroreception and electrogenesis in fishes, with a focus on freshwater taxa and with emphasis on the proximate (morphological, physiological and genetic) bases of EOD and electroreceptor diversity. Second, to describe the diversity, biogeography, ecology and electric signal diversity of the mormyroids and gymnotiforms and to explore the ultimate (evolutionary) bases of signal and receptor diversity in their convergent electrogenic-electrosensory systems. Four sets of potential drivers or moderators of signal diversity are discussed. First, selective forces of an abiotic (environmental) nature for optimal electrolocation and communication performance of the EOD. Second, selective forces of a biotic nature targeting the communication function of the EOD, including sexual selection, reproductive interference from syntopic heterospecifics and selection from eavesdropping predators. Third, non-adaptive drift and, finally, phylogenetic inertia, which may arise from stabilising selection for optimal signal-receptor matching.

Stem-Cell-Driven Growth and Regrowth of the Adult Spinal Cord in Teleost Fish.

The spinal cord of the teleost fish Apteronotus leptorhynchus continues to grow during adulthood, in concert with the overall body growth. Immunohistological studies, combined with mathematical modeling, suggest that this growth is driven by proliferative activity of Sox2-expressing stem/progenitor cells (SPCs) and by cell drift due to population pressure. The SPCs exhibit high volumetric density in the caudal filament and the ependymal layer. Nevertheless, the majority of these cells are found in the parenchyma throughout the rostro-caudal axis of the spinal cord, albeit at much lower volumetric densities than in the ependymal layer. The SPCs give rise, via transit-amplifying cells, to neurons and glia. The relative number of neurons and glia is primarily regulated through apoptosis of supernumerary neurons. Quantitative analysis has demonstrated that the continued cell proliferation results in additive neurogenesis. This addition includes adult-born spinal electromotoneurons, thereby resulting in a continuous increase in the amplitude of the fish's electric organ discharge during adult life. Amputation of the caudal part of the spinal cord induces initially a degenerative response, dominated by massive apoptotic cell death in spinal cord tissue immediately rostral to the injury site, and distinguished by a partial loss of the electric organ discharge amplitude. This phase is followed by a regenerative response, characterized by absence of gliosis and by rapid stem-cell-driven tissue regrowth. Although the quality of the regenerated tissue is variable among individuals, the structural repair has led in every fish examined thus far to full recovery of the electric organ discharge amplitude.

Molecular phylogeny of the ghost knifefishes (Gymnotiformes: Apteronotidae)

Ghost knifefishes (Gymnotiformes: Apteronotidae) are weakly electric fishes that possess a high-frequency, neurogenic electric organ discharge. They are found throughout the humid Neotropics from Panama to Argentina and are most diverse and abundant in the channels of large lowland rivers. Apteronotidae is the most species-rich family of Neotropical electric knifefishes with 96 valid species in 15 genera. We present a phylogenetic hypothesis based on molecular sequence data from three mitochondrial genes (16S, coi, cytb) and four nuclear loci (glyt, rag2, ryr3, zic1). Our analysis includes sequence data for 203 samples in 54 species and 14 genera, making it the most densely-sampled and data-rich phylogeny of the Apteronotidae to date. Our results corroborate previous phylogenetic hypotheses with the placement of Orthosternarchus + Sternarchorhamphus sister to all other apteronotids, a non-monophyletic Apterontotus, and a sister relationship between Sternarchorhynchus and the Navajini. We also report several novel relationships, particularly within the Navajini and among several species of the nominal genus Apteronotus not previously included in phylogenetic analyses. We additionally provide a new classification for the family.

Neuronal Dynamics Underlying Communication Signals in a Weakly Electric Fish: Implications for Connectivity in a Pacemaker Network

Neuronal networks can produce stable oscillations and synchrony that are under tight control yet flexible enough to rapidly switch between dynamical states. The pacemaker nucleus in the weakly electric fish comprises a network of electrically coupled neurons that fire synchronously at high frequency. This activity sets the timing for an oscillating electric organ discharge with the lowest cycle-to-cycle variability of all known biological oscillators. Despite this high temporal precision, pacemaker activity is behaviorally modulated on millisecond time-scales for the generation of electrocommunication signals. The network mechanisms that allow for this combination of stability and flexibility are not well understood. In this study, we use an in vitro pacemaker preparation from Apteronotus leptorhynchus to characterize the neural responses elicited by the synaptic inputs underlying electrocommunication. These responses involve a variable increase in firing frequency and a prominent desynchronization of neurons that recovers within 5 oscillation cycles. Using a previously developed computational model of the pacemaker network, we show that the frequency changes and rapid resynchronization observed experimentally are most easily explained when model neurons are interconnected more densely and with higher coupling strengths than suggested by published data. We suggest that the pacemaker network achieves both stability and flexibility by balancing coupling strength with interconnectivity and that variation in these network features may provide a substrate for species-specific evolution of electrocommunication signals.