Electrosensory and metabolic responses of weakly electric fish to changing water conductivity.

Weakly electric gymnotiform fishes use self-generated electric organ discharges (EODs) to navigate and communicate. The electrosensory range for these processes is a function of EOD amplitude, determined by the fish's electric organ (EO) output and the electrical conductivity of the surrounding water. Anthropogenic activity, such as deforestation, dams and industrial/agricultural runoff, are known to increase water conductivity in neotropical habitats, likely reducing the electrosensory range of these fish. We investigated whether fish modulate EO output as means of re-expanding electrosensory range after a rapid increase in water conductivity in the pulse-type Brachyhypopomus gauderio and the wave-type Eigenmannia virescens. Furthermore, because EOD production incurs significant metabolic costs, we assessed whether such compensation is associated with an increase in metabolic rate. Following the conductivity increase, B. gauderio increased EOD amplitude by 20.2±4.3% over 6 days but with no associated increase in metabolic rate, whereas the EOD amplitude of E. virescens remained constant, accompanied by an unexpected decrease in metabolic rate. Our results suggest that B. gauderio uses a compensation mechanism that requires no metabolic investment, such as impedance matching, or a physiological trade-off wherein energy is diverted from other physiological processes to increase EO output. These divergent responses between species could be the result of differences in reproductive life history or evolutionary adaptations to different aquatic habitats. Continued investigation of electrosensory responses to changing water conditions will be essential for understanding the effects of anthropogenic disturbances on gymnotiforms, and potential physiological mechanisms for adapting to a rapidly changing aquatic environment.

Sensorimotor adaptation to destabilizing dynamics in weakly electric fish.

Humans and other animals can readily learn to compensate for changes in the dynamics of movement. Such changes can result from an injury or changes in the weight of carried objects. These changes in dynamics can lead not only to reduced performance but also to dramatic instabilities. We evaluated the impacts of compensatory changes in control policies in relation to stability and robustness in Eigenmannia virescens, a species of weakly electric fish. We discovered that these fish retune their sensorimotor control system in response to experimentally generated destabilizing dynamics. Specifically, we used an augmented reality system to manipulate sensory feedback during an image stabilization task in which a fish maintained its position within a refuge. The augmented reality system measured the fish's movements in real time. These movements were passed through a high-pass filter and multiplied by a gain factor before being fed back to the refuge motion. We adjusted the gain factor to gradually destabilize the fish's sensorimotor loop. The fish retuned their sensorimotor control system to compensate for the experimentally induced destabilizing dynamics. This retuning was partially maintained when the augmented reality feedback was abruptly removed. The compensatory changes in sensorimotor control improved tracking performance as well as control-theoretic measures of robustness, including reduced sensitivity to disturbances and improved phase margins.

Understanding daily rhythms in weakly electric fish: the role of melatonin on the electric behavior of Brachyhypopomus gauderio.

Living organisms display molecular, physiological and behavioral rhythms synchronized with natural environmental cycles. Understanding the interaction between environment, physiology and behavior requires taking into account the complexity of natural habitats and the diversity of behavioral and physiological adaptations. Brachyhypopomus gauderio is characterized by the emission of electric organ discharges (EOD), with a very stable rate modulated by social and environmental cues. The nocturnal arousal in B. gauderio coincides with a melatonin-dependent EOD rate increase. Here, we first show a daily cycle in both the EOD basal rate (EOD-BR) and EOD-BR variability of B. gauderio in nature. We approached the understanding of the role of melatonin in this natural behavior through both behavioral pharmacology and in vitro assays. We report, for the first time in gymnotiformes, a direct effect of melatonin on the pacemaker nucleus (PN) in in vitro preparation. Melatonin treatment lowered EOD-BR in freely moving fish and PN basal rate, while increasing the variability of both. These results show that melatonin plays a key role in modulating the electric behavior of B. gauderio through its effect on rate and variability, both of which must be under a tight temporal regulation to prepare the animal for the challenging nocturnal environment.

The sensory effects of light on the electric organ discharge rate of Gymnotus omarorum.

Gymnotiformes are nocturnal fishes inhabiting the root mats of floating plants. They use their electric organ discharge (EOD) to explore the environment and to communicate. Here, we show and describe tonic and phasic sensory-electromotor responses to light distinct from indirect effects depending on the light-induced endogenous circadian rhythm. In the dark, principally during the night, inter-EOD interval histograms are bimodal: the main peak corresponds to the basal rate and a secondary peak corresponds to high-frequency bouts. Light causes a twofold tonic but opposing effect on the EOD histogram: (i) decreasing the main mode and (ii) blocking the high-frequency bouts and consequently increasing the main peak at the expense of removal of the secondary one. Additionally, light evokes phasic responses whose amplitude increases with intensity but whose slow time course and poor adaptation differentiate from the so-called novelty responses evoked by abrupt changes in sensory stimuli of other modalities. We confirmed that Gymnotus omarorum tends to escape from light, suggesting that these phasic responses are probably part of a global 'light-avoidance response'. We interpret the data within an ecological context. Fish rest under the shade of aquatic plants during the day and light spots due to the sun's relative movement alert the fish to hide in shady zones to avoid macroptic predators and facilitate tracking the movement of floating plant islands by wind and/or water currents.

The effect of urethane and MS-222 anesthesia on the electric organ discharge of the weakly electric fish Apteronotus leptorhynchus.

Urethane and MS-222 are agents widely employed for general anesthesia, yet, besides inducing a state of unconsciousness, little is known about their neurophysiological effects. To investigate these effects, we developed an in vivo assay using the electric organ discharge (EOD) of the weakly electric fish Apteronotus leptorhynchus as a proxy for the neural output of the pacemaker nucleus. The oscillatory neural activity of this brainstem nucleus drives the fish's EOD in a one-to-one fashion. Anesthesia induced by urethane or MS-222 resulted in pronounced decreases of the EOD frequency, which lasted for up to 3 h. In addition, each of the two agents caused a manifold increase in the generation of transient modulations of the EOD known as chirps. The reduction in EOD frequency can be explained by the modulatory effect of urethane on neurotransmission, and by the blocking of voltage-gated sodium channels by MS-222, both within the circuitry controlling the neural oscillations of the pacemaker nucleus. The present study demonstrates a marked effect of urethane and MS-222 on neural activity within the central nervous system and on the associated animal's behavior. This calls for caution when conducting neurophysiological experiments under general anesthesia and interpreting their results.

Proteomic characterization of spontaneously regrowing spinal cord following injury in the teleost fish Apteronotus leptorhynchus, a regeneration-competent vertebrate.

In adult mammals, spontaneous repair after spinal cord injury (SCI) is severely limited. By contrast, teleost fish successfully regenerate injured axons and produce new neurons from adult neural stem cells after SCI. The molecular mechanisms underlying this high regenerative capacity are largely unknown. The present study addresses this gap by examining the temporal dynamics of proteome changes in response to SCI in the brown ghost knifefish (Apteronotus leptorhynchus). Two-dimensional difference gel electrophoresis (2D DIGE) was combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and tandem mass spectrometry (MS/MS) to collect data during early (1 day), mid (10 days), and late (30 days) phases of regeneration following caudal amputation SCI. Forty-two unique proteins with significant differences in abundance between injured and intact control samples were identified. Correlation analysis uncovered six clusters of spots with similar expression patterns over time and strong conditional dependences, typically within functional families or between isoforms. Significantly regulated proteins were associated with axon development and regeneration; proliferation and morphogenesis; neuronal differentiation and re-establishment of neural connections; promotion of neuroprotection, redox homeostasis, and membrane repair; and metabolism or energy supply. Notably, at all three time points examined, significant regulation of proteins involved in inflammatory responses was absent.

Electrocommunication signals and aggressive behavior vary among male morphs in an apteronotid fish, Compsaraia samueli.

Within-species variation in male morphology is common among vertebrates and is often characterized by dramatic differences in behavior and hormonal profiles. Males with divergent morphs also often use communication signals in a status-dependent way. Weakly electric knifefish are an excellent system for studying variation in male morphology and communication and its hormonal control. Knifefish transiently modulate the frequency of their electric organ discharge (EOD) during social encounters to produce chirps and rises. In the knifefish Compsaraia samueli, males vary extensively in jaw length. EODs and their modulations (chirps and rises) have never been investigated in this species, so it is unclear whether jaw length is related to the function of these signals. We used three behavioral assays to analyze EOD modulations in male C. samueli: (1) artificial playbacks, (2) relatively brief, live agonistic dyadic encounters, and (3) long-term overnight recordings. We also measured circulating levels of two androgens, 11-ketotestosterone and testosterone. Chirp structure varied within and across individuals in response to artificial playback, but was unrelated to jaw length. Males with longer jaws were more often dominant in dyadic interactions. Chirps and rises were correlated with and preceded attacks regardless of status, suggesting these signals function in aggression. In longer-term interactions, chirp rate declined after 1 week of pairing, but was unrelated to male morphology. Levels of circulating androgens were low and not predictive of jaw length or EOD signal parameters. These results suggest that communication signals and variation in male morphology are linked to outcomes of non-breeding agonistic contests.

Glutamatergic control of a pattern-generating central nucleus in a gymnotiform fish.

The activity of central pattern-generating networks (CPGs) may change under the control exerted by various neurotransmitters and modulators to adapt its behavioral outputs to different environmental demands. Although the mechanisms underlying this control have been well established in invertebrates, most of their synaptic and cellular bases are not yet well understood in vertebrates. Gymnotus omarorum, a pulse-type gymnotiform electric fish, provides a well-suited vertebrate model to investigate these mechanisms. G. omarorum emits rhythmic and stereotyped electric organ discharges (EODs), which function in both perception and communication, under the command of an electromotor CPG. This nucleus is composed of electrotonically coupled intrinsic pacemaker cells, which pace the rhythm, and bulbospinal projecting relay cells that contribute to organize the pattern of the muscle-derived effector activation that produce the EOD. Descending influences target CPG neurons to produce adaptive behavioral electromotor responses to different environmental challenges. We used electrophysiological and pharmacological techniques in brainstem slices of G. omarorum to investigate the underpinnings of the fast transmitter control of its electromotor CPG. We demonstrate that pacemaker, but not relay cells, are endowed with ionotropic and metabotropic glutamate receptor subtypes. We also show that glutamatergic control of the CPG likely involves two types of synapses contacting pacemaker cells, one type containing both α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) and N-methyl-d-aspartate (NMDA) receptors and the other one only-NMDA receptor. Fast neurotransmitter control of vertebrate CPGs seems to exploit the kinetics of the involved postsynaptic receptors to command different behavioral outputs. The prospect of common neural designs to control CPG activity in vertebrates is discussed.NEW & NOTEWORTHY Underpinnings of neuromodulation of central pattern-generating networks (CPG) have been well characterized in many species. The effects of fast neurotransmitter systems remain, however, poorly understood. This research uses in vitro electrophysiological and pharmacological techniques to show that the neurotransmitter control of a vertebrate CPG in gymnotiform fish involves the convergence of only-NMDA and AMPA-NMDA glutamatergic synapses onto neurons that pace the rhythm. These inputs may organize different behavioral outputs according to their distinct functional properties.

Simultaneous spike-time locking to multiple frequencies.

Locking of neural firing is ubiquitously observed in the brain and occurs when neurons fire at a particular phase or in synchronization with an external signal. Here we study in detail the locking of single neurons to multiple distinct frequencies at the example of p-type electroreceptor afferents in the electrosensory system of the weakly electric fish Apteronotus leptorhynchus (brown ghost knifefish). We find that electrosensory afferents and pyramidal cells in the electrosensory lateral line lobe (ELL) lock to multiple frequencies, including the electric organ discharge (EOD) frequency, beat, and stimulus itself. We identify key elements necessary for locking to multiple frequencies, study its limits, and provide concise mathematical models reproducing our main findings. Our findings provide another example of how rate and temporal codes can coexist and complement each other in single neurons and demonstrate that sensory coding in p-type electroreceptor afferents provides a much richer representation of the sensory environment than commonly assumed. Since the underlying mechanisms are not specific to the electrosensory system, our results could provide the basis for studying multiple frequency locking in other systems.NEW & NOTEWORTHY Locking of neuronal spikes to external and internal signals is a ubiquitous neurophysiological mechanism that has been extensively studied in several brain areas and species. Using experimental data from the electrosensory system and concise mathematical models, we analyze how a single neuron can simultaneously lock to multiple frequencies. Our findings demonstrate how temporal and rate codes can complement each other and lead to rich neuronal representations of sensory signals.

Reduced brain cell proliferation following somatic injury is buffered by social interaction in electric fish, Apteronotus leptorhynchus.

In many species, the negative effects of aversive stimuli are mitigated by social interactions, a phenomenon termed social buffering. In one form of social buffering, social interactions reduce the inhibition of brain cell proliferation during stress. Indirect predator stimuli (e.g., olfactory or visual cues) are known to decrease brain cell proliferation, but little is known about how somatic injury, as might occur from direct predator encounter, affects brain cell proliferation and whether this response is influenced by conspecific interactions. Here, we assessed the social buffering of brain cell proliferation in an electric fish, Apteronotus leptorhynchus, by examining the separate and combined effects of tail injury and social interactions. We mimicked a predator-induced injury by amputating the caudal tail tip, exposed fish to paired interactions that varied in timing, duration and recovery period, and measured brain cell proliferation and the degree of social affiliation. Paired social interaction mitigated the negative effects of tail amputation on cell proliferation in the forebrain but not the midbrain. Social interaction either before or after tail amputation reduced the effect of tail injury and continuous interaction both before and after caused an even greater buffering effect. Social interaction buffered the proliferation response after short-term (1 d) or long-term recovery (7 d) from tail amputation. This is the first report of social buffering of brain cell proliferation in a non-mammalian model. Despite the positive association between social stimuli and brain cell proliferation, we found no evidence that fish affiliate more closely following tail injury.

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.

Waveform discrimination in a pair of pulse-generating electric fishes.

Studies of pulse-type gymnotiform electric fishes have suggested that electric organ discharge waveforms (EODw) allow individuals to discriminate between conspecific and allospecific signals, but few have approached this experimentally. Here we implement a phase-locked playback technique for a syntopic species pair, Brachyhypopomus gauderio and Gymnotus omarorum. Both species respond to changes in stimulus waveform with a transitory reduction in the interpulse interval of their self-generated discharge, providing strong evidence of discrimination. We also document sustained rate changes in response to different EODws, which may suggest recognition of natural waveforms.

Variability in locomotor dynamics reveals the critical role of feedback in task control.

Animals vary considerably in size, shape, and physiological features across individuals, but yet achieve remarkably similar behavioral performances. We examined how animals compensate for morphophysiological variation by measuring the system dynamics of individual knifefish (Eigenmannia virescens) in a refuge tracking task. Kinematic measurements of Eigenmannia were used to generate individualized estimates of each fish's locomotor plant and controller, revealing substantial variability between fish. To test the impact of this variability on behavioral performance, these models were used to perform simulated 'brain transplants'-computationally swapping controllers and plants between individuals. We found that simulated closed-loop performance was robust to mismatch between plant and controller. This suggests that animals rely on feedback rather than precisely tuned neural controllers to compensate for morphophysiological variability.

People come in different shapes and sizes, but most will perform similarly well if asked to complete a task requiring fine manual dexterity ­ such as holding a pen or picking up a single grape. How can different individuals, with different sized hands and muscles, produce such similar movements? One explanation is that an individual's brain and nervous system become precisely tuned to mechanics of the body's muscles and skeleton. An alternative explanation is that brain and nervous system use a more "robust" control policy that can compensate for differences in the body by relying on feedback from the senses to guide the movements. To distinguish between these two explanations, Uyanik et al. turned to weakly electric freshwater fish known as glass knifefish. These fish seek refuge within root systems, reed grass and among other objects in the water. They swim backwards and forwards to stay hidden despite constantly changing currents. Each fish shuttles back and forth by moving a long ribbon-like fin on the underside of its body. Uyanik et al. measured the movements of the ribbon fin under controlled conditions in the laboratory, and then used the data to create computer models of the brain and body of each fish. The models of each fish's brain and body were quite different. To study how the brain interacts with the body, Uyanik et al. then conducted experiments reminiscent of those described in the story of Frankenstein and transplanted the brain from each computer model into the body of different model fish. These "brain swaps" had almost no effect on the model's simulated swimming behavior. Instead, these "Frankenfish" used sensory feedback to compensate for any mismatch between their brain and body. This suggests that, for some behaviors, an animal's brain does not need to be precisely tuned to the specific characteristics of its body. Instead, robust control of movement relies on many seemingly redundant systems that provide sensory feedback. This has implications for the field of robotics. It further suggests that when designing robots, engineers should prioritize enabling the robots to use sensory feedback to cope with unexpected events, a well-known idea in control engineering.

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.

Strategies of object polarization and their role in electrosensory information gathering.

Weakly electric fish polarize the nearby environment with a stereotyped electric field and gain information by detecting the changes imposed by objects with tuned sensors. Here we focus on polarization strategies as paradigmatic bioinspiring mechanisms for sensing devices. We begin this research developing a toy model that describes three polarization strategies exhibited by three different groups of fish. We then report an experimental analysis which confirmed predictions of the model and in turn predicted functional consequences that were explored in behavioral experiments in the pulse fish Gymnotus omarorum. In the experiments, polarization was evaluated by estimating the object's stamp (i.e. the electric source that produces the same electric image as the object) as a function of object impedance, orientation, and position. Signal detection and discrimination was explored in G. omarorum by provoking novelty responses, which are known to reflect the increment in the electric image provoked by a change in nearby impedance. To achieve this, we stepped the longitudinal impedance of a cylindrical object between two impedances (either capacitive or resistive). Object polarization and novelty responses indicate that G. omarorum has two functional regions in the electrosensory field. At the front of the fish, there is a foveal field where object position and orientation are encoded in signal intensity, while the qualia associated with impedance is encoded in signal time course. On the side of the fish there is a peripheral field where the complexity of the polarizing field facilitates detection of objects oriented in any angle with respect to the fish´s longitudinal axis. These findings emphasize the importance of articulating field generation, sensor tuning and the repertoire of exploratory movements to optimize performance of artificial active electrosensory systems.

Hormone-mediated modulation of the electromotor CPG in pulse-type weakly electric fish. Commonalities and differences across species.

Like stomatogastric activity in crustaceans, vocalization in teleosts and frogs, and locomotion in mammals, the electric organ discharge (EOD) of weakly electric fish is a rhythmic and stereotyped electromotor pattern. The EOD, which functions in both perception and communication, is controlled by a two-layered central pattern generator (CPG), the electromotor CPG, which modifies its basal output in response to environmental and social challenges. Despite major anatomo-functional commonalities in the electromotor CPG across electric fish species, we show that Gymnotus omarorum and Brachyhypopomus gauderio have evolved divergent neural processes to transiently modify the CPG outputs through descending fast neurotransmitter inputs to generate communication signals. We also present two examples of electric behavioral displays in which it is possible to separately analyze the effects of neuropeptides (mid-term modulation) and gonadal steroid hormones (long-term modulation) upon the CPG. First, the nonbreeding territorial aggression of G. omarorum has been an advantageous model to analyze the status-dependent modulation of the excitability of CPG neuronal components by vasotocin. Second, the seasonal and sexually dimorphic courtship signals of B. gauderio have been useful to understand the effects of sex steroids on the responses to glutamatergic inputs in the CPG. Overall, the electromotor CPG functions in a regime that safeguards the EOD waveform. However, prepacemaker influences and hormonal modulation enable an enormous versatility and allows the EOD to adapt its functional state in a species-, sex-, and social context-specific manners.

A shocking discovery of threat risks on newly described species of weakly electric fishes.

This study aims to investigate relationships between species traits and publication date in the weakly electric osteoglossiform Mormyroidea (African knifefish and elephantfishes) and the ostariophysan Gymnotiformes (Neotropical knifefishes). It is investigated whether body size and geographic distribution area are correlated with publication date and whether extinction risk differs between both phylogenetically distant and geographically isolated clades. Statistical modelling indicates that the number of new species described annually is stable in mormyroids and clearly increasing in gymnotiforms. Best-fitting generalised linear models (GLM) indicate that the newly discovered species are more often of small-bodied, predominantly narrowly distributed and more likely to be threatened with extinction. These characteristics are more pronounced in mormyroids when compared with gymnotiforms, suggesting that some African electric fishes may live an ephemeral existence after formal description. Despite taxonomic work has been more intense in the Neotropics than in Africa in the recent decades, there is evidence that the African continent represents the next frontier of species descriptions. Taxonomic studies are fundamental for the understanding of richness and distribution and hence extinction risk assessment and conservation, of these remarkable convergent fish clades.

Cellular and Network Mechanisms May Generate Sparse Coding of Sequential Object Encounters in Hippocampal-Like Circuits.

The localization of distinct landmarks plays a crucial role in encoding new spatial memories. In mammals, this function is performed by hippocampal neurons that sparsely encode an animal's location relative to surrounding objects. Similarly, the dorsolateral pallium (DL) is essential for spatial learning in teleost fish. The DL of weakly electric gymnotiform fish receives both electrosensory and visual input from the preglomerular nucleus (PG), which has been hypothesized to encode the temporal sequence of electrosensory or visual landmark/food encounters. Here, we show that DL neurons in the Apteronotid fish and in the Carassius auratus (goldfish) have a hyperpolarized resting membrane potential (RMP) combined with a high and dynamic spike threshold that increases following each spike. Current-evoked spikes in DL cells are followed by a strong small-conductance calcium-activated potassium channel (SK)-mediated after-hyperpolarizing potential (AHP). Together, these properties prevent high frequency and continuous spiking. The resulting sparseness of discharge and dynamic threshold suggest that DL neurons meet theoretical requirements for generating spatial memory engrams by decoding the landmark/food encounter sequences encoded by PG neurons. Thus, DL neurons in teleost fish may provide a promising, simple system to study the core cell and network mechanisms underlying spatial memory.

Calbindin-D28k expression in spinal electromotoneurons of the weakly electric fish Apteronotus leptorhynchus during adult development and regeneration.

Additive neurogenesis, the net increase in neuronal numbers by addition of new nerve cells to existing tissue, forms the basis for indeterminate spinal cord growth in brown ghost knifefish (Apteronotus leptorhynchus). Among the cells generated through the activity of adult neural stem cells are electromotoneurons, whose axons constitute the electric organ of this weakly electric fish. Electromotoneuron development is organized along a caudo-rostral gradient, with the youngest and smallest of these cells located near the caudal end of the spinal cord. Electromotoneurons start expressing calbindin-D28k when their somata have reached diameters of approximately 10 µm, and they continue expression after they have grown to a final size of about 50 µm. Calbindin-D28k expression is significantly increased in young neurons generated in response to injury. Immunohistochemical staining against caspase-3 revealed that electromotoneurons in both intact and regenerating spinal cord are significantly less likely to undergo apoptosis than the average spinal cord cell. We hypothesize that expression of calbindin-D28k protects electromotoneurons from cell death; and that the evolutionary development of such a neuroprotective mechanism has been driven by the indispensability of electromotoneurons in the fish's electric behavior, and by the high size-dependent costs associated with their production or removal upon cell death.

Distinctive mechanisms underlie the emission of social electric signals of submission in Gymnotus omarorum.

South American weakly electric fish (order Gymnotiformes) rely on a highly conserved and relatively fixed electromotor circuit to produce species-specific electric organ discharges (EODs) and a variety of meaningful adaptive EOD modulations. The command for each EOD arises from a medullary pacemaker nucleus composed of electrotonically coupled intrinsic pacemaker and bulbospinal projecting relay cells. During agonistic encounters, Gymnotus omarorum signals submission by interrupting its EOD (offs) and emitting transient high-rate barrages of low-amplitude discharges (chirps). Previous studies in Gymnotiformes have shown that electric signal diversity is based on the segregation of descending synaptic inputs to pacemaker or relay cells and differential activation of the neurotransmitter receptors -for glutamate or γ-aminobutyric acid (GABA) - of these cells. Therefore, we tested whether GABAergic and glutamatergic inputs to pacemaker nucleus neurons are involved in the emission of submissive electric signals in G. omarorum We found that GABA applied to pacemaker cells evokes EOD interruptions that closely resemble natural offs. Although in other species chirping is probably due to glutamatergic suprathreshold depolarization of relay cells, here, application of glutamate to these cells was unable to replicate the emission of this submissive signal. Nevertheless, chirp-like discharges were emitted after the enhancement of excitability of relay cells by blocking an IA-type potassium current and, in some cases, by application of vasotocin, a status-dependent modulator peptide of G. omarorum agonistic behavior. Modulation of the electrophysiological properties of pacemaker nucleus neurons in Gymnotiformes emerges as a novel putative mechanism endowing electromotor networks with higher functional versatility.