Active sensing associated with spatial learning reveals memory-based attention in an electric fish.

Active sensing behaviors reveal what an animal is attending to and how it changes with learning. Gymnotus sp, a gymnotiform weakly electric fish, generates an electric organ discharge (EOD) as discrete pulses to actively sense its surroundings. We monitored freely behaving gymnotid fish in a large dark "maze" and extracted their trajectories and EOD pulse pattern and rate while they learned to find food with electrically detectable landmarks as cues. After training, they more rapidly found food using shorter, more stereotyped trajectories and spent more time near the food location. We observed three forms of active sensing: sustained high EOD rates per unit distance (sampling density), transient large increases in EOD rate (E-scans) and stereotyped scanning movements (B-scans) were initially strong at landmarks and food, but, after learning, intensified only at the food location. During probe (no food) trials, after learning, the fish's search area and intense active sampling was still centered on the missing food location, but now also increased near landmarks. We hypothesize that active sensing is a behavioral manifestation of attention and essential for spatial learning; the fish use spatial memory of landmarks and path integration to reach the expected food location and confine their attention to this region.

Ontogeny of the electric organs in the electric eel, Electrophorus electricus: physiological, histological, and fine structural investigations.

This study attempts to clarify the controversy regarding the ontogenetic origin of the main organ electrocytes in the electric eel, Electrophorus electricus. The dispute was between an earlier claimed origin from a skeletal muscle precursor [Fritsch, 1881], or from a distinct electrocyte-generating matrix, or germinative zone [Keynes, 1961]. We demonstrate electrocyte formation from a metamerically organized group of pre-electroblasts, splitting off the ventralmost tip of the embryonic trunk mesoderm at the moment of hatching from the egg. We show details of successive stages in the development of rows of electric plates, the electrocytes, by means of conventional histology and electron microscopy. The membrane-bound pre-electroblasts multiply rapidly and then undergo a specific mitosis where they lose their membranes and begin extensive cytoplasm production as electroblasts. Electrical activity, consisting of single and multiple pulses, was noticed in seven-day-old larvae that began to exhibit swimming movements. A separation of discharges into single pulses and trains of higher voltage pulses was seen first in 45-mm-long larvae. A lateralis imus muscle and anal fin ray muscles, implicated by earlier investigators in the formation of electrocytes, begin developing at a time in larval life when eight columns of electrocytes are already present. Axonal innervation is seen very early during electrocyte formation.

Computational modeling of electric imaging in weakly electric fish: insights for physiology, behavior and evolution.

Weakly electric fish can sense electric signals produced by other animals whether they are conspecifics, preys or predators. These signals, sensed by passive electroreception, sustain electrocommunication, mating and agonistic behavior. Weakly electric fish can also generate a weak electrical discharge with which they can actively sense the animate and inanimate objects in their surroundings. Understanding both sensory modalities depends on our knowledge of how pre-receptorial electric images are formed and how movements modify them during behavior. The inability of effectively measuring pre-receptorial fields at the level of the skin contrasts with the amount of knowledge on electric fields and the availability of computational methods for estimating them. In this work we review past work on modeling of electric organ discharge and electric images, showing the usefulness of these methods to calculate the field and providing a brief explanation of their principles. In addition, we focus on recent work demonstrating the potential of electric image modeling and what the method has to offer for experimentalists studying sensory physiology, behavior and evolution.

The slow pathway in the electrosensory lobe of Gymnotus omarorum: field potentials and unitary activity.

This is a first communication on the self-activation pattern of the electrosensory lobe in the pulse weakly electric fish Gymnotus omarorum. Field potentials in response to the fish's own electric organ discharge (EOD) were recorded along vertical tracks (50µm step) and on a transversal lattice array across the electrosensory lobe (resolution 50µm×100µm). The unitary activity of 82 neurons was recorded in the same experiments. Field potential analysis indicates that the slow electrosensory path shows a characteristic post-EOD pattern of activity marked by three main events: (i) a small and early component at about 7ms, (ii) an intermediate peak about 13ms and (iii) a late broad component peaking after 20ms. Unit firing rate showed a wide range of latencies between 3 and 30ms and a variable number of spikes (median 0.28units/EOD). Conditional probability analysis showed monomodal and multimodal post-EOD histograms, with the peaks of unit activity histograms often matching the timing of the main components of the field potentials. Monomodal responses were sub-classified as phase locked monomodal (variance smaller than 1ms), early monomodal (intermediate variance, often firing in doublets, peaking range 10-17ms) and late monomodal (large variance, often firing two spikes separated about 10ms, peaking beyond 17ms). The responses of multimodal units showed that their firing probability was either enhanced, or depressed just after the EOD. In this last (depressed) subtype of unit the probability stepped down just after the EOD. Early inhibition and the presence of early phase locked units suggest that the observed pattern may be influenced by a fast feed forward inhibition. We conclude that the ELL in pulse gymnotiformes is activated in a complex sequence of events that reflects the ELL network connectivity.

Local vasotocin modulation of the pacemaker nucleus resembles distinct electric behaviors in two species of weakly electric fish.

The neural bases of social behavior diversity in vertebrates have evolved in close association with hypothalamic neuropeptides. In particular, arginine-vasotocin (AVT) is a key integrator underlying differences in behavior across vertebrate taxa. Behavioral displays in weakly electric fish are channeled through specific patterns in their electric organ discharges (EODs), whose rate is ultimately controlled by a medullary pacemaker nucleus (PN). We first explored interspecific differences in the role of AVT as modulator of electric behavior in terms of EOD rate between the solitary Gymnotus omarorum and the gregarious Brachyhypopomus gauderio. In both species, AVT IP injection (10µg/gbw) caused a progressive increase of EOD rate of about 30%, which was persistent in B. gauderio, and attenuated after 30min in G. omarorum. Secondly, we demonstrated by in vitro electrophysiological experiments that these behavioral differences can be accounted by dissimilar effects of AVT upon the PN in itself. AVT administration (1µM) to the perfusion bath of brainstem slices containing the PN produced a small and transient increase of PN activity rate in G. omarorum vs the larger and persistent increase previously reported in B. gauderio. We also identified AVT neurons, for the first time in electric fish, using immunohistochemistry techniques and confirmed the presence of hindbrain AVT projections close to the PN that might constitute the anatomical substrate for AVT influences on PN activity. Taken together, our data reinforce the view of the PN as an extremely plastic medullary central pattern generator that not only responds to higher influences to adapt its function to diverse contexts, but also is able to intrinsically shape its response to neuropeptide actions, thus adding a hindbrain target level to the complexity of the global integration of central neuromodulation of electric behavior.

Static frequency tuning accounts for changes in neural synchrony evoked by transient communication signals.

Although communication signals often vary continuously on the underlying signal parameter, they are perceived as distinct categories. We here report the opposite case where an electrocommunication signal is encoded in four distinct regimes, although the behavior described to date does not show distinct categories. In particular, we studied the encoding of chirps by P-unit afferents in the weakly electric fish Apteronotus leptorhynchus. These fish generate an electric organ discharge that oscillates at a certain individual-specific frequency. The interaction of two fish in communication contexts leads to the emergence of a beating amplitude modulation (AM) at the frequency difference between the two individual signals. This frequency difference represents the social context of the encounter. Chirps are transient increases of the fish's frequency leading to transient changes in the frequency of the AM. We stimulated the cells with the same chirp on different, naturally occurring backgrounds beats. The P-units responded either by synchronization or desynchronization depending on the background. Although the duration of a chirp is often shorter than a full cycle of the AM it elicits, the distinct responses of the P-units to the chirp can be predicted solely from the frequency of the AM based on the static frequency tuning of the cells.

Sensory processing and corollary discharge effects in posterior caudal lobe Purkinje cells in a weakly electric mormyrid fish.

Although it has been suggested that the cerebellum functions to predict the sensory consequences of motor commands, how such predictions are implemented in cerebellar circuitry remains largely unknown. A detailed and relatively complete account of predictive mechanisms has emerged from studies of cerebellum-like sensory structures in fish, suggesting that comparisons of the cerebellum and cerebellum-like structures may be useful. Here we characterize electrophysiological response properties of Purkinje cells in a region of the cerebellum proper of weakly electric mormyrid fish, the posterior caudal lobe (LCp), which receives the same mossy fiber inputs and projects to the same target structures as the electrosensory lobe (ELL), a well-studied cerebellum-like structure. We describe patterns of simple spike and climbing fiber activation in LCp Purkinje cells in response to motor corollary discharge, electrosensory, and proprioceptive inputs and provide evidence for two functionally distinct Purkinje cell subtypes within LCp. Protocols that induce rapid associative plasticity in ELL fail to induce plasticity in LCp, suggesting differences in the adaptive functions of the two structures. Similarities and differences between LCp and ELL are discussed in light of these results.

Dendritic SK channels convert NMDA-R-dependent LTD to burst timing-dependent plasticity.

Feedback and descending projections from higher to lower brain centers play a prominent role in all vertebrate sensory systems. Feedback might be optimized for the specific sensory processing tasks in their target brain centers, but it has been difficult to connect the properties of feedback synapses to sensory tasks. Here, we use the electrosensory system of a gymnotiform fish (Apteronotus leptorhynchus) to address this problem. Cerebellar feedback to pyramidal cells in the first central electrosensory processing region, the electrosensory lateral line lobe (ELL), is critical for canceling spatially and temporally redundant electrosensory input. The ELL contains four electrosensory maps, and we have previously analyzed the synaptic and network bases of the redundancy reduction mechanism in a map (centrolateral segment; CLS) believed to guide electrolocation behavior. In the CLS, only long-term depression was induced by pairing feedback presynaptic and pyramidal cell postsynaptic bursts. In this paper, we turn to an ELL map (lateral segment; LS) known to encode electrocommunication signals. We find remarkable differences in synaptic plasticity of the morphologically identical cerebellar feedback input to the LS. In the LS, pyramidal cell SK channels permit long-term potentiation (LTP) of feedback synapses when pre- and postsynaptic bursts occur at the same time. We hypothesize that LTP in this map is required for enhancing the encoding of weak electrocommunication signals. We conclude that feedback inputs that appear morphologically identical in sensory maps dedicated to different tasks, nevertheless display different synaptic plasticity rules contributing to differential sensory processing in these maps.

Electric organ discharge diversity in the genus Gymnotus: anatomo-functional groups and electrogenic mechanisms.

Previous studies describe six factors accounting for interspecific diversity of electric organ discharge (EOD) waveforms in Gymnotus. At the cellular level, three factors determine the locally generated waveforms: (1) electrocyte geometry and channel repertoire; (2) the localization of synaptic contacts on electrocyte surfaces; and (3) electric activity of electromotor axons preceding the discharge of electrocytes. At the organismic level, three factors determine the integration of the EOD as a behavioral unit: (4) the distribution of different types of electrocytes and specialized passive tissue forming the electric organ (EO); (5) the neural mechanisms of electrocyte discharge coordination; and (6) post-effector mechanisms. Here, we reconfirm the importance of the first five of these factors based on comparative studies of a wider diversity of Gymnotus than previously investigated. Additionally, we report a hitherto unseen aspect of EOD diversity in Gymnotus. The central region of the EO (which has the largest weight on the conspecific-received field) usually exhibits a negative-positive-negative pattern where the delay between the early negative and positive peaks (determined by neural coordination mechanisms) matches the delay between the positive and late negative peaks (determined by electrocyte responsiveness). Because delays between peaks typically determine the peak power frequency, this matching implies a co-evolution of neural and myogenic coordination mechanisms in determining the spectral specificity of the intraspecific communication channel. Finally, we define four functional species groups based on EO/EOD structure. The first three exhibit a heterogeneous EO in which doubly innervated electrocytes are responsible for a main triphasic complex. Group I species exhibit a characteristic cephalic extension of the EO. Group II species exhibit an early positive component of putative neural origin, and strong EO auto-excitability. Group III species exhibit an early, slow, negative wave of abdominal origin, and variation in EO auto-excitability. Representatives of Group IV generate a unique waveform comprising a main positive peak followed by a small, load-dependent negative component.

Receptive field organization across multiple electrosensory maps. II. Computational analysis of the effects of receptive field size on prey localization.

The electric fish Apteronotus leptorhynchus emits a high-frequency electric organ discharge (EOD) sensed by specialized electroreceptors (P-units) distributed across the fish's skin. Objects such as prey increase the amplitude of the EOD over the underlying skin and thus cause an increase in P-unit discharge. The resulting localized intensity increase is called the electric image and is detected by its effect on the P-unit population; the electric image peak value and the extent to its spreads are cues utilized by these fish to estimate the location and size of its prey. P-units project topographically to three topographic maps in the electrosensory lateral line lobe (ELL): centromedial (CMS), centrolateral (CLS), and lateral (LS) segments. In a companion paper I have calculated the receptive fields (RFs) in these maps: RFs were small in CMS and very large in LS, with intermediate values in CLS. Here I use physiological data to create a simple model of the RF structure within the three ELL maps and to compute the response of these model maps to simulated prey. The Fisher information (FI) method was used to compute the optimal estimates possible for prey localization across the three maps. The FI predictions were compared with behavioral studies on prey detection. These comparisons were used to frame alternative hypotheses on the functions of the three maps and on the constraints that RF size and synaptic strength impose on weak signal detection and estimation.

Receptive field organization across multiple electrosensory maps. I. Columnar organization and estimation of receptive field size.

The electric fish Apteronotus leptorhynchus emits a high-frequency electric organ discharge (EOD) sensed by specialized electroreceptors (P-units). Amplitude modulations (AMs) of the EOD are caused by objects such as prey as well as by social interactions with conspecifics. The firing rate of P-units is modulated by the AMs due to both objects and communication signals. P-units trifurcate as they enter the medulla; they terminate topographically with three maps of the electrosensory lateral line lobe (ELL): the centromedial (CMS), centrolateral (CLS), and lateral (LS) segments. Within each map P-units terminate onto the basal dendrites of pyramidal cells. Anterograde filling of P-units and retrograde filling of the basal bushes of pyramidal cells were used to estimate their respective spreads and spacing in the three maps. These estimates were used to compute the receptive field structure of the pyramidal cells: receptive fields were small in CMS and very large in LS with intermediate values in CLS. There are several classes of pyramidal cells defined by morphological and functional criteria; these cells are organized into columns such that each column contains one member of each class and all cells within a column receive the same P-unit input.

Waveform generation in the weakly electric fish Gymnotus coropinae (Hoedeman): the electric organ and the electric organ discharge.

This article deals with the electric organ and its discharge in Gymnotus coropinae, a representative species of one of the three main clades of the genus. Three regions with bilateral symmetry are described: (1) subopercular (medial and lateral columns of complex shaped electrocytes); (2) abdominal (medial and lateral columns of cuboidal and fusiform electrocytes); and (3) main [four columns, one dorso-lateral (containing fusiform electrocytes) and three medial (containing cuboidal electrocytes)]. Subopercular electrocytes are all caudally innervated whereas two of the medial subopercular ones are also rostrally innervated. Fusiform electrocytes are medially innervated at the abdominal portion, and at their rostral and caudal poles at the main portion. Cuboidal electrocytes are always caudally innervated. The subopercular portion generates a slow head-negative wave (V(1r)) followed by a head-positive spike (V(3r)). The abdominal and main portions generate a fast tetra-phasic complex (V(2345ct)). Since subopercular components prevail in the near field and the rest in the far field, time coincidence of V(3r) with V(2) leads to different waveforms depending on the position of the receiver. This confirms the splitting hypothesis of communication and exploration channels based on the different timing, frequency band and reach of the regional waveforms. The following hypothesis is compatible with the observed anatomo-functional organization: V(1r) corresponds to the rostral activation of medial subopercular electrocytes and V(3r) to the caudal activation of all subopercular electrocytes; V(2), and part of V(3ct), corresponds to the successive activation of the rostral and caudal poles of dorso-lateral fusiform electrocytes; and V(345ct) is initiated in the caudal face of cuboidal electrocytes by synaptic activation (V(3ct)) and it is completed (V(45ct)) by the successive activation of rostral and caudal faces by the action currents evoked in the opposite face.

Active electroreception in Gymnotus omari: imaging, object discrimination, and early processing of actively generated signals.

Weakly electric fishes "electrically illuminate" the environment in two forms: pulse fishes emit a succession of discrete electric discharges while wave fishes emit a continuous wave. These strategies are present in both taxonomic groups of weakly electric fishes, mormyrids and gymnotids. As a consequence one can distinguish four major types of active electrosensory strategies evolving in parallel. Pulse gymnotids have an electrolocating strategy common with pulse mormyrids, but brains of pulse and wave gymnotids are alike. The beating strategy associated to other differences in the electrogenic system and electrosensory responses suggests that similar hardware might work in a different mode for processing actively generated electrosensory images. In this review we summarize our findings in pulse gymnotids' active electroreception and outline a primary agenda for the next research.

Population coding by electrosensory neurons.

Sensory stimuli typically activate many receptors at once and therefore should lead to increases in correlated activity among central neurons. Such correlated activity could be a critical feature in the encoding and decoding of information in central circuits. Here we characterize correlated activity in response to two biologically relevant classes of sensory stimuli in the primary electrosensory nuclei, the electrosensory lateral line lobe, of the weakly electric fish Apteronotus leptorhynchus. Our results show that these neurons can display significant correlations in their baseline activities that depend on the amount of receptive field overlap. A detailed analysis of spike trains revealed that correlated activity resulted predominantly from a tendency to fire synchronous or anti-synchronous bursts of spikes. We also explored how different stimulation protocols affected correlated activity: while prey-like stimuli increased correlated activity, conspecific-like stimuli decreased correlated activity. We also computed the correlations between the variabilities of each neuron to repeated presentations of the same stimulus (noise correlations) and found lower amounts of noise correlation for communication stimuli. Therefore the decrease in correlated activity seen with communication stimuli is caused at least in part by reduced noise correlations. This differential modulation in correlated activity occurred because of changes in burst firing at the individual neuron level. Our results show that different categories of behaviorally relevant input will differentially affect correlated activity. In particular, we show that the number of correlated bursts within a given time window could be used by postsynaptic neurons to distinguish between both stimulus categories.

Sensory processing in the fast electrosensory pathway of pulse gymnotids studied at multiple integrative levels.

Pulse gymnotids extract information about the environment using the pulsed discharge of an electric organ. Cutaneous electroreceptor organs transduce and encode the changes that objects imprint on the self-generated transcutaneous electric field. This review deals with the role of a neural circuit, the fast electrosensory path of pulse gymnotids, in the streaming of self generated electrosensory signals. The activation of this path triggers a low-responsiveness window slightly shorter than the interval between electric organ discharges. This phenomenon occurs at the electrosensory lateral line lobe where primary afferent terminals project on the somata of spherical neurons. The main subservient mechanism of the low-responsiveness window rely on the intrinsic properties of spherical neurons (dominated by a voltage dependent, low-threshold, non-inactivating and slowly-deactivating K(+) conductance) determining the cell to respond with a single spike followed by a long refractory period. Externally generated signals that randomly occur within the interval between self-generated discharges are likely blocked by the low responsiveness window. Repetitive signals, as those emitted by conspecifics with a slightly lower rate, occur progressively at longer delays beyond the duration of the low responsiveness window. Transient increases of the discharge rate relocate the interference within the low-responsiveness window. We propose that this combination of sensory filtering and electromotor control favors the self-generated signals in detriment of other, securing the continuity of the electrolocation stream.

Stimulus selectivity is enhanced by voltage-dependent conductances in combination-sensitive neurons.

Central sensory neurons often respond selectively to particular combinations of stimulus attributes, but we know little about the underlying cellular mechanisms. The weakly electric fish Gymnarchus discriminates the sign of the frequency difference (Df) between a neighbor's electric organ discharge (EOD) and its own EOD by comparing temporal patterns of amplitude modulation (AM) and phase modulation (PM). Sign-selective neurons in the midbrain respond preferentially to either positive frequency differences (Df >0 selective) or negative frequency differences (Df <0 selective). To study the mechanisms of combination sensitivity, we made whole cell intracellular recordings from sign-selective midbrain neurons in vivo and recorded postsynaptic potential (PSP) responses to AM, PM, Df >0, and Df <0. Responses to AM and PM consisted of alternating excitatory and inhibitory PSPs. These alternating responses were in phase for the preferred sign of Df and offset for the nonpreferred sign of Df. Therefore a certain degree of sign selectivity was predicted by a linear sum of the responses to AM and PM. Responses to the nonpreferred sign of Df, but not the preferred sign of Df, were substantially weaker than linear predictions, causing a significant increase in the actual degree of sign selectivity. By using various levels of current clamp and comparing our results to simple models of synaptic integration, we demonstrate that this decreased response to the nonpreferred sign of Df is caused by a reduction in voltage-dependent excitatory conductances. This finding reveals that nonlinear decoders, in the form of voltage-dependent conductances, can enhance the selectivity of single neurons for particular combinations of stimulus attributes.

Ambiguous encoding of stimuli by primary sensory afferents causes a lack of independence in the perception of multiple stimulus attributes.

Accurate sensory perception often depends on the independent encoding and subsequent integration of multiple stimulus attributes. In the weakly electric fish Eigenmannia, P- and T-type primary afferent fibers are specialized for encoding the amplitude and phase, respectively, of electrosensory stimuli. We used a stimulus estimation technique to quantify the ability of P- and T-units to encode random modulations in amplitude and phase. As expected, P-units exhibited a clear preference for encoding amplitude modulations, whereas T-units exhibited a clear preference for encoding phase modulations. Surprisingly, both types of afferents also encoded their nonpreferred stimulus attribute when it was presented in isolation or when the preferred stimulus attribute was sufficiently weak. Because afferent activity can be affected by modulations in either amplitude or phase, it is not possible to unambiguously distinguish between these two stimulus attributes by observing the activity of a single afferent fiber. Simple model neurons with a preference for encoding either amplitude or phase also encoded their nonpreferred stimulus attribute when it was presented in isolation, suggesting that such ambiguity is unavoidable. Using the well known jamming avoidance response as a probe of electrosensory perception, we show that the ambiguity at the single-neuron level gives rise to a systematic misrepresentation of stimuli at the population level and a resulting misperception of the amplitude and phase of electrosensory stimuli.

Immunocytochemical identification of cell types in the mormyrid electrosensory lobe.

The electrosensory lobes (ELLs) of mormyrid and gymnotid fish are useful sites for studying plasticity and descending control of sensory processing. This study used immunocytochemistry to examine the functional circuitry of the mormyrid ELL. We used antibodies against the following proteins and amino acids: the neurotransmitters glutamate and gamma-aminobutyric acid (GABA); the GABA-synthesizing enzyme glutamic acid decarboxylase (GAD); GABA transporter 1; the anchoring protein for GABA and glycine receptors, gephyrin; the calcium binding proteins calbindin and calretinin; the NR1 subunit of the N-methyl-D-aspartate glutamate receptor; the metabotropic glutamate receptors mGluR1alpha, mGluR2/3, and mGluR5; and the intracellular signaling molecules calcineurin, calcium calmodulin kinase IIalpha (CAMKIIalpha) and the receptor for inositol triphosphate (IP3R1alpha). Selective staining allowed for identification of new cell types including a deep granular layer cell that relays sensory information from primary afferent fibers to higher order cells of ELLS. Selective staining also allowed for estimates of relative numbers of different cell types. Dendritic staining of Purkinje-like medium ganglion cells with antibodies against metabotropic glutamate receptors and calcineurin suggests hypotheses concerning mechanisms of the previously demonstrated synaptic plasticity in these cells. Finally, several cell types including the above-mentioned granular cells, thick-smooth dendrite cells, and large multipolar cells of the intermediate layer were present in the two zones of ELL that receive input from mormyromast electroreceptors but were absent in the zone of ELL that receives input from ampullary electroreceptors, indicating markedly different processing for these two types of input. J. Comp. Neurol. 483:124-142, 2005. (c) 2005 Wiley-Liss, Inc.

L-citrulline immunoreactivity reveals nitric oxide production in the electromotor and electrosensory systems of the weakly electric fish, Apteronotus leptorhynchus.

Weakly electric fish produce electric organ discharges (EODs) used for electrolocation and communication. In the brown ghost knifefish, Apteronotus leptorhynchus, several neuron types in brain regions that control the EOD or process electrosensory information express nitric oxide synthase (NOS). The present study used immunoreactivity for L-citrulline, a byproduct of the production of nitric oxide (NO) by NOS, to assess NO production in NOS-expressing neurons. A polyclonal antibody against L-citrulline produced specific labeling in most neuronal populations previously identified to express NOS. Specifically, several cell types that precisely encode temporal information and/or fire at high frequencies, including spherical cells in the electrosensory lateral line lobe, giant cells in layer VI of the dorsal torus semicircularis, and pacemaker and relay cells in the pacemaker nucleus, were strongly immunoreactive for L-citrulline. This suggests that these neurons produced high levels of NO. Notably, electromotor neurons, which also strongly express NOS, were not immunoreactive for L-citrulline, suggesting that NOS did not produce high levels of NO in these neurons. No apparent differences in L-citrulline distribution or intensity were observed between socially isolated fish and fish exposed to playback stimuli simulating the presence of a conspecific. This suggests that social stimulation by electrocommunication signals is not necessary for high levels of NO production in many NOS-positive neurons. Future studies focusing on regulation of NO production in these systems, and the effects of NO on electrosensory processing and electromotor pattern generation will help elucidate the function of NO signaling pathways in this system.

Sex and species differences in neuromodulatory input to a premotor nucleus: a comparative study of substance P and communication behavior in weakly electric fish.

Many electric fish species modulate their electric organ discharges (EODs) to produce transient social signals that vary in number and structure. In Apteronotus leptorhynchus, males modulate their EOD more often than females, whereas in Apteronotus albifrons, males and females produce similar numbers of modulations. Sex differences in the number of EOD modulations in A. leptorhynchus are associated with sex differences in substance P in the diencephalic nucleus that controls transient EOD modulations, the CP/PPn. These sex differences in substance P have been hypothesized to regulate sex differences in the production of EOD modulations. To comparatively test this hypothesis, we examined substance P immunoreactivity in the CP/PPn of male and female A. leptorhynchus and A. albifrons. Because the number of EOD modulations is sexually monomorphic in A. albifrons, we predicted no sex difference in substance P in the CP/PPn of this species. Contrary to this prediction, male A. albifrons had significantly more substance P in the CP/PPn than females. This suggests that sex differences in substance P are not sufficient for controlling sex differences in the number of EOD modulations. Modulation structure (frequency excursion and/or duration), however, is also sexually dimorphic in A. leptorhynchus and is another possible behavioral correlate of the sexually dimorphic distribution of substance P. The present study found pronounced sex differences in the structure of EOD modulations in A. albifrons similar to those in A. leptorhynchus. Thus, sex differences in substance P may influence sex differences in the structure, rather than the number, of EOD modulations.