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.

The weakly electric fish, Apteronotus albifrons, actively avoids experimentally induced hypoxia.

Anthropogenic environmental degradation has led to an increase in the frequency and prevalence of aquatic hypoxia (low dissolved oxygen concentration, DO), which may affect habitat quality for water-breathing fishes. The weakly electric black ghost knifefish, Apteronotus albifrons, is typically found in well-oxygenated freshwater habitats in South America. Using a shuttle-box design, we exposed juvenile A. albifrons to a stepwise decline in DO from normoxia (> 95% air saturation) to extreme hypoxia (10% air saturation) in one compartment and chronic normoxia in the other. On average, A. albifrons actively avoided the hypoxic compartment below 22% air saturation. Hypoxia avoidance was correlated with upregulated swimming activity. Following avoidance, fish regularly ventured back briefly into deep hypoxia. Hypoxia did not affect the frequency of their electric organ discharges. Our results show that A. albifrons is able to sense hypoxia at non-lethal levels and uses active avoidance to mitigate its adverse effects.

Androgens regulate sex differences in signaling but are not associated with male variation in morphology in the weakly electric fish Parapteronotus hasemani.

Sexually dimorphic signaling is widespread among animals and can act as an honest indicator of mate quality. Additionally, differences in signaling and morphology within a sex can be associated with different strategies for acquiring mates. Weakly electric fish communicate via self-generated electrical fields that transmit information about sex, reproductive state, and social status. The weakly electric knifefish Parapteronotus hasemani exhibits sexual dimorphism in body size as well as substantial within-male variation in body size and jaw length. We asked whether P. hasemani exhibits hormonally mediated sexual dimorphism in electrocommunication behavior. We also asked whether males with short versus long jaws differed significantly from each other in morphology, behavior, hormone levels, or reproductive maturity. Males produced longer chirps than females, but other signal parameters (electric organ discharge frequency; chirp rate and frequency modulation) were sexually monomorphic. Pharmacologically blocking androgen receptors in males reduced chirp duration, suggesting that this sexually dimorphic trait is regulated at least in part by the activational effects of androgens. Males sorted into two distinct morphological categories but did not differ in circulating 11-ketotestosterone or testosterone. Short-jawed males and long-jawed males also did not differ in any aspects of signaling. Thus, chirping and high levels of 11-ketotestosterone were reliably associated with reproductively active males but do not necessarily indicate male type or quality. This contrasts with other alternative male morph systems in which males that differ in morphology also differ in androgen profiles and signaling behavior.

Melatonin Regulates Daily Variations in Electric Behavior Arousal in Two Species of Weakly Electric Fish with Different Social Structures.

Timing is crucial for social interactions. Animal behavior is synchronized with biotic and abiotic environment variables ensuring that the activity phase of conspecifics occurs during the same period of the day. As biological rhythms are embedded in the complex integrative control of the brain, it is fundamental to explore its interaction with environmental and social factors. This approach will unravel the link between external stimuli carrying information on environmental cycles and the neural commands for behavior, including social behavior, associated with precise phases of those cycles. Arousal in the solitary Gymnotus omarorum and in the gregarious Brachyhypopomus gauderio is characterized by a nocturnal increase in the basal discharge rate of electric behavior, which is mild and transient in G. omarorum and large and persistent in B. gauderio. In this study, we show that the major integrator of social behavior, AVT (arginine vasotocin), is not involved in the nocturnal increase of electric behavior basal rate in isolated animals of either species. On the other hand, endogenous melatonin, the major modulator of the circadian system, is responsible for the nocturnal increase in electric behavior in isolated individuals of both species.

A highly polarized excitable cell separates sodium channels from sodium-activated potassium channels by more than a millimeter.

The bioelectrical properties and resulting metabolic demands of electrogenic cells are determined by their morphology and the subcellular localization of ion channels. The electric organ cells (electrocytes) of the electric fish Eigenmannia virescens generate action potentials (APs) with Na(+) currents >10 µA and repolarize the AP with Na(+)-activated K(+) (KNa) channels. To better understand the role of morphology and ion channel localization in determining the metabolic cost of electrocyte APs, we used two-photon three-dimensional imaging to determine the fine cellular morphology and immunohistochemistry to localize the electrocytes' ion channels, ionotropic receptors, and Na(+)-K(+)-ATPases. We found that electrocytes are highly polarized cells ∼ 1.5 mm in anterior-posterior length and ∼ 0.6 mm in diameter, containing ∼ 30,000 nuclei along the cell periphery. The cell's innervated posterior region is deeply invaginated and vascularized with complex ultrastructural features, whereas the anterior region is relatively smooth. Cholinergic receptors and Na(+) channels are restricted to the innervated posterior region, whereas inward rectifier K(+) channels and the KNa channels that terminate the electrocyte AP are localized to the anterior region, separated by >1 mm from the only sources of Na(+) influx. In other systems, submicrometer spatial coupling of Na(+) and KNa channels is necessary for KNa channel activation. However, our computational simulations showed that KNa channels at a great distance from Na(+) influx can still terminate the AP, suggesting that KNa channels can be activated by distant sources of Na(+) influx and overturning a long-standing assumption that AP-generating ion channels are restricted to the electrocyte's posterior face.

Effects of in vitro exposure to mercury on male gonads and sperm structure of the tropical fish tuvira Gymnotus carapo (L.).

This study investigated the progressive effects of HgCl2 in the testis and sperm of the tropical fish tuvira Gymnotus carapo L. exposed to increasing Hg concentrations (5-30 µm) and increasing exposure times (24-96 h). Histopathology and metal concentrations in the testis were observed. Hg concentrations in the testis reached 5.1 and 5.2 µg g(-1) in fish exposed to 20 and 30 µm of Hg, respectively. Hg effects on testicular tissue were observed even at low Hg concentrations, with no alterations in gonadosomatic index. However, the quantitative analysis of the induced alterations (lesion index) demonstrated that the Hg effects in testis became more severe after exposure to higher concentrations (20 and 30 µm) and during longer exposure (72 and 96 h), probably leading to partial or total loss of the organ function. Hg exposure (20 µm) also affected sperm count and altered sperm morphology. This study showed that HgCl2 caused progressive damage to testicular tissue, reduced sperm count and altered sperm morphology. These results are important in establishing a direct correlation between Hg accumulation and severity of lesions.

Tight hormonal phenotypic integration ensures honesty of the electric signal of male and female Brachyhypopomus gauderio.

Hormones mediate sexually selected traits including advertisement signals. Hormonal co-regulation links the signal to other hormonally-mediated traits such that the tighter the integration, the more reliable the signal is as a predictor of those other traits. Androgen administration increases the duration of the communication signal pulse in both sexes of the electric fish Brachyhypopomus gauderio. To determine whether the duration of the signal pulse could function as an honest indicator of androgen levels and other androgen-mediated traits, we measured the variation in sex steroids, signal pulse duration, and sexual development throughout the breeding season of B. gauderio in marshes in Uruguay. Although the sexes had different hormone titres and signal characteristics, in both sexes circulating levels of the androgens testosterone (T) and 11-ketotestosterone (11-KT) were strongly related to signal pulse duration. Consequently, signal pulse duration can serve as an honest indicator of circulating androgens in males and females alike. Additionally, through phenotypic integration, signal pulse duration also predicts other sexual traits directly related to androgen production: gonad size in males and estradiol (E2) levels in females. Our findings show that tight hormonal phenotypic integration between advertisement signal and other sex steroid-mediated traits renders the advertisement signal an honest indicator of a suite of reproductive traits.

Environmental complexity, seasonality and brain cell proliferation in a weakly electric fish, Brachyhypopomus gauderio.

Environmental complexity and season both influence brain cell proliferation in adult vertebrates, but their relative importance and interaction have not been directly assessed. We examined brain cell proliferation during both the breeding and non-breeding seasons in adult male electric fish, Brachyhypopomus gauderio, exposed to three environments that differed in complexity: (1) a complex natural habitat in northern Uruguay, (2) an enriched captive environment where fish were housed socially and (3) a simple laboratory setting where fish were isolated. We injected fish with BrdU 2.5 h before sacrifice to label newborn cells. We examined the hindbrain and midbrain and quantified the density of BrdU+ cells in whole transverse sections, proliferative zones and two brain nuclei in the electrocommunication circuitry (the pacemaker nucleus and the electrosensory lateral line lobe). Season had the largest effect on cell proliferation, with fish during the breeding season having three to seven times more BrdU+ cells than those during the non-breeding season. Although the effect was smaller, fish from a natural environment had greater rates of cell proliferation than fish in social or isolated captive environments. For most brain regions, fish in social and isolated captive environments had equivalent levels of cell proliferation. However, for brain regions in the electrocommunication circuitry, group-housed fish had more cell proliferation than isolated fish, but only during the breeding season (season × environment interaction). The regionally and seasonally specific effect of social environment on cell proliferation suggests that addition of new cells to these nuclei may contribute to seasonal changes in electrocommunication behavior.

Energetic cost of communication.

Communication signals may be energetically expensive or inexpensive to produce, depending on the function of the signal and the competitive nature of the communication system. Males of sexually selected species may produce high-energy advertisement signals, both to enhance detectability and to signal their size and body condition. Accordingly, the proportion of the energy budget allocated to signal production ranges from almost nothing for many signals to somewhere in excess of 50% for acoustic signals in short-lived sexually selected species. Recent data from gymnotiform electric fish reveal mechanisms that regulate energy allocated to sexual advertisement signals through dynamical remodeling of the excitable membranes in the electric organ. Further, males of the short-lived sexually selected species, Brachyhypopomus gauderio, trade off among different metabolic compartments, allocating energy to signal production while reducing energy used in other metabolic functions. Female B. gauderio, by contrast, do not trade off energy between signaling and other functions. To fuel energetically expensive signal production, we expect a continuum of strategies to be adopted by animals of different life history strategies. Future studies should explore the relation between life history and energy allocation trade-offs.

Energetic constraints on electric signalling in wave-type weakly electric fishes.

Gymnotiform weakly electric fishes generate electric organ discharges (EODs) and sense perturbations of the resulting electric field for purposes of orientation, prey detection and communication. Some species produce oscillatory ('wave-type') EODs at very high frequencies (up to 2 kHz) that have been proposed to be energetically expensive. If high-frequency EODs are expensive, then fish may modulate their EOD frequency and/or amplitude in response to low-oxygen (hypoxic) stress and/or compensate for costs of signalling through other adaptations that maximize oxygen uptake efficiency. To test for evidence of an energetic cost of signalling, we recorded EOD in conjunction with metabolic rates, critical oxygen tension and aquatic surface respiration (ASR(90)) thresholds in Apteronotus leptorhynchus, a species found in high-oxygen habitats, and Eigenmannia virescens, a species more typically found in low-oxygen waters. Eigenmannia virescens had a lower mean ASR(90) threshold and critical oxygen tension compared with A. leptorhynchus, consistent with field distributions. Within each species, there was no evidence for a relationship between metabolic rate and either EOD frequency or amplitude under normoxia, suggesting that there is no significant direct metabolic cost associated with producing a higher frequency EOD. However, when exposed to progressive hypoxia, fish generally responded by reducing EOD amplitude, which may reduce energetic costs. The threshold at which fish reduced EOD amplitude tended to be lower in E. virescens, a pattern consistent with higher tolerance to hypoxic stress. The results of this study suggest that wave-type fish reduce their EOD amplitude to reduce direct energetic costs without reducing metabolic rate under hypoxia.

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.

Serotonergic activation of 5HT1A and 5HT2 receptors modulates sexually dimorphic communication signals in the weakly electric fish Apteronotus leptorhynchus.

Serotonin modulates agonistic and reproductive behavior across vertebrate species. 5HT(1A) and 5HT(1B) receptors mediate many serotonergic effects on social behavior, but other receptors, including 5HT(2) receptors, may also contribute. We investigated serotonergic regulation of electrocommunication signals in the weakly electric fish Apteronotus leptorhynchus. During social interactions, these fish modulate their electric organ discharges (EODs) to produce signals known as chirps. Males chirp more than females and produce two chirp types. Males produce high-frequency chirps as courtship signals; whereas both sexes produce low-frequency chirps during same-sex interactions. Serotonergic innervation of the prepacemaker nucleus, which controls chirping, is more robust in females than males. Serotonin inhibits chirping and may contribute to sexual dimorphism and individual variation in chirping. We elicited chirps with EOD playbacks and pharmacologically manipulated serotonin receptors to determine which receptors regulated chirping. We also asked whether serotonin receptor activation generally modulated chirping or more specifically targeted particular chirp types. Agonists and antagonists of 5HT(1B/1D) receptors (CP-94253 and GR-125743) did not affect chirping. The 5HT(1A) receptor agonist 8OH-DPAT specifically increased production of high-frequency chirps. The 5HT(2) receptor agonist DOI decreased chirping. Receptor antagonists (WAY-100635 and MDL-11939) opposed the effects of their corresponding agonists. These results suggest that serotonergic inhibition of chirping may be mediated by 5HT(2) receptors, but that serotonergic activation of 5HT(1A) receptors specifically increases the production of high-frequency chirps. The enhancement of chirping by 5HT(1A) receptors may result from interactions with cortisol and/or arginine vasotocin, which similarly enhance chirping and are influenced by 5HT(1A) activity in other systems.

Voltage-gated potassium conductances in Gymnotus electrocytes(AB).

Electrocytes are muscle-derived cells that generate the electric organ discharge (EOD) in most gymnotiform fish. We used an in vitro preparation to determine if the complex EOD of Gymnotus carapo was related to the membrane properties of electrocytes. We discovered that in addition to the three Na(+)-mediated conductances described in a recent paper [Sierra F, Comas V, Buño W, Macadar O (2005) Sodium-dependent plateau potentials in electrocytes of the electric fish Gymnotus carapo. J Comp Physiol A 191:1-11] there were four K(+)-dependent conductances. Membrane depolarization activated a delayed rectifier (I(K)) and an A-type (I(A)) current. I(A) displayed fast voltage-dependent activation-inactivation kinetics, was blocked by 4-aminopyridine (1 mM) and played a major role in action potential (AP) repolarization. Its voltage dependence and kinetics shape the brief AP that typifies Gymnotus electrocytes. The I(K) activated by depolarization contributed less to AP repolarization. Membrane hyperpolarization uncovered two inward rectifiers (IR1 and IR2) with voltage dependence and kinetics that correspond to the complex "hyperpolarizing responses" (HRs) described under current-clamp. IR1 shows "instantaneous" activation, is blocked by Ba(2+) and Cs(+) and displays a voltage and time dependent inactivation that matches the hyperpolarizing phase of the HR. The activation of IR2 is slower and at more negative potentials than IR1 and is resistant to Ba(2+) and Cs(+). This current fits the depolarizing phase of the HR. The EOD waveform of Gymnotus carapo is more complex than that of other gymnotiform fish species, the complexity originates in the voltage responses generated through the interactions of three Na(+) and four K(+) voltage- and time-dependent conductances although the innervation pattern also contributes [Trujillo-Cenóz O, Echagüe JA (1989) Waveform generation of the electric organ discharge in Gymnotus carapo. I. Morphology and innervation of the electric organ. J Comp Physiol A 165:343-351].

Subsecond Sensory Modulation of Serotonin Levels in a Primary Sensory Area and Its Relation to Ongoing Communication Behavior in a Weakly Electric Fish.

Serotonergic neurons of the raphe nuclei of vertebrates project to most regions of the brain and are known to significantly affect sensory processing. The subsecond dynamics of sensory modulation of serotonin levels and its relation to behavior, however, remain unknown. We used fast-scan cyclic voltammetry to measure serotonin release in the electrosensory system of weakly electric fish, Apteronotus leptorhynchus. These fish use an electric organ to generate a quasi-sinusoidal electric field for communicating with conspecifics. In response to conspecific signals, they frequently produce signal modulations called chirps. We measured changes in serotonin concentration in the hindbrain electrosensory lobe (ELL) with a resolution of 0.1 s concurrently with chirping behavior evoked by mimics of conspecific electric signals. We show that serotonin release can occur phase locked to stimulus onset as well as spontaneously in the ELL region responsible for processing these signals. Intense auditory stimuli, on the other hand, do not modulate serotonin levels in this region, suggesting modality specificity. We found no significant correlation between serotonin release and chirp production on a trial-by-trial basis. However, on average, in the trials where the fish chirped, there was a reduction in serotonin release in response to stimuli mimicking similar-sized same-sex conspecifics. We hypothesize that the serotonergic system is part of an intricate sensory-motor loop: serotonin release in a sensory area is triggered by sensory input, giving rise to motor output, which can in turn affect serotonin release at the timescale of the ongoing sensory experience and in a context-dependent manner.

Connections between the central posterior/prepacemaker nucleus and hypothalamic areas in the weakly electric fish Apteronotus leptorhynchus: evidence for an indirect, but not a direct, link.

The central posterior/prepacemaker nucleus (CP/PPn) of the weakly electric fish Apteronotus leptorhynchus consists of a few thousands of neurons in the dorsal thalamus. Subpopulations of this complex play a crucial role in neural control of transient modulations of the otherwise extremely constant electric organ discharges. Because both the propensity to execute these modulations and the type of modulations produced may vary enormously with the behavioral situation, it has been hypothesized that this behavioral plasticity is, partially, mediated by peptidergic neuromodulators originating from hypothalamic areas. To define the structural basis of this proposed modulatory input, we have in the present study examined the connections between the CP/PPn proper and hypothalamic areas by employing an in vitro tract-tracing technique. Neither anterograde nor retrograde tracing experiments could provide evidence for the existence of a direct link between the CP/PPn proper and hypothalamic areas. However, the results of our investigation suggest an indirect connection between the CP/PPn proper and two hypothalamic regions, the hypothalamus ventralis and the hypothalamus lateralis, with the preglomerular nucleus serving as a relay station.

Resource use by two electric fishes (Gymnotiformes) of the National Forest Saracá-Taquera, Oriximiná, Pará

Fishes of the order Gymnotiformes have high diversity of oral and head morphology, which suggests trophic specializations within each clade. The aim of this study was to describe resource use patterns by two fish species (Gymnorhamphichthys rondoni and Gymnotus coropinae) in the National Forest Saracá-Taquera, Oriximiná - Pará, analyzing microhabitat use, diet composition, feeding strategies, niche breadth and niche overlap. Stomach contents of 101 individuals (41 G. rondoni and 60 G. coropinae), sampled in 23 headwater streams were analyzed and volume of food items was quantified to characterize their feeding ecology. Gymnorhamphichthys rondoni was captured mainly on sandy bottoms, whereas G. coropinae in crevices. Both species had a zoobenthivorous diet and consumed predominantly Sediment/Detritus and Diptera larvae, but also included allochthonous prey in their diet. These species had high niche overlap, with small variations related to the higher consumption of Ceratopogonidae larvae by G. rondoni and of Chironomidae larvae by G. coropinae. Both species had a generalist feeding strategy, but G. coropinae had a broader niche breadth. Our results demonstrate that G. rondoni and G. coropinae occupy different microhabitats but rely on similar food resources.(AU)

Peixes da ordem Gymnotiformes apresentam alta diversidade morfológica, o que sugere a existência de especializações tróficas dentro dos clados. O objetivo deste estudo foi analisar o uso de recursos por duas espécies de peixes elétricos (Gymnorhamphichthys rondoni e Gymnotus coropinae) na Floresta Nacional Saracá-Taquera, Oriximiná - Pará, analisando o uso de microhabitats, composição da dieta, estratégias alimentares, amplitude de nicho e sobreposição de nicho. Conteúdos estomacais de 101 indivíduos (41 G. rondoni e 60 G. coropinae), capturados em 23 igarapés de cabeceira, foram analisados e quantificados volumetricamente para a caracterização da ecologia trófica. Gymnorhamphichthys rondoni foi registrada principalmente em bancos de areia, enquanto G. coropinae em fendas. Ambas as espécies apresentaram dieta zoobentívora e consumiram predominantemente sedimento/detritos e larvas de Diptera, mas também consumiram presas alóctones. Foi observada alta sobreposição de nicho trófico e a baixa variação encontrada foi relacionada à maior utilização de larvas de Chironomidae por G. rondoni e de larvas de Ceratopogonidae por G. coropinae. Ambas as espécies apresentaram estratégia alimentar generalista, porém G. coropinae apresentou maior amplitude de nicho. Nossos resultados demonstram que G. rondoni e G. coropinae ocupam microhabitats diferentes, mas dependem de recursos alimentares similares.(AU)

Fish geometry and electric organ discharge determine functional organization of the electrosensory epithelium.

Active electroreception in Gymnotus omarorum is a sensory modality that perceives the changes that nearby objects cause in a self generated electric field. The field is emitted as repetitive stereotyped pulses that stimulate skin electroreceptors. Differently from mormyriformes electric fish, gymnotiformes have an electric organ distributed along a large portion of the body, which fires sequentially. As a consequence shape and amplitude of both, the electric field generated and the image of objects, change during the electric pulse. To study how G. omarorum constructs a perceptual representation, we developed a computational model that allows the determination of the self-generated field and the electric image. We verify and use the model as a tool to explore image formation in diverse experimental circumstances. We show how the electric images of objects change in shape as a function of time and position, relative to the fish's body. We propose a theoretical framework about the organization of the different perceptive tasks made by electroreception: 1) At the head region, where the electrosensory mosaic presents an electric fovea, the field polarizing nearby objects is coherent and collimated. This favors the high resolution sampling of images of small objects and perception of electric color. Besides, the high sensitivity of the fovea allows the detection and tracking of large faraway objects in rostral regions. 2) In the trunk and tail region a multiplicity of sources illuminate different regions of the object, allowing the characterization of the shape and position of a large object. In this region, electroreceptors are of a unique type and capacitive detection should be based in the pattern of the afferents response. 3) Far from the fish, active electroreception is not possible but the collimated field is suitable to be used for electrocommunication and detection of large objects at the sides and caudally.

Expression of myogenic regulatory factors in the muscle-derived electric organ of Sternopygus macrurus.

In most groups of electric fish, the current-producing cells of electric organs (EOs) derive from striated muscle fibers but retain some phenotypic characteristics of their precursor muscle cells. Given the role of the MyoD family of myogenic regulatory factors (MRFs) in the transcriptional activation of the muscle program in vertebrates, we examined their expression in the electrocytes of the gymnotiform Sternopygus macrurus. We estimated the number of MRF genes in the S. macrurus genome and our Southern blot analyses revealed a single MyoD, myogenin, myf5 and MRF4 gene. Quantitative RT-PCR showed that muscle and EO transcribe all MRF genes. With the exception of MyoD, the endogenous levels of myogenin, myf5 and MRF4 transcripts in electrocytes were greater than those detected in muscle fibers. These data indicate that MRF expression levels are not sufficient to predict the level to which the muscle program is manifested. Qualitative expression analysis of MRF co-regulators MEF2C, Id1 and Id2 also revealed these genes not to be unique to either muscle or EO, and detected similar expression patterns in the two tissues. Therefore, the partial muscle program of the EO is not associated with a partial expression of MRFs or with apparent distinct levels of some MRF co-factors. In addition, electrical inactivation by spinal cord transection (ST) resulted in the up-regulation of some muscle proteins in electrocytes without an accompanying increase in MRF transcript levels or notable changes in the co-factors MEF2C, Id1 and Id2. These findings suggest that the neural regulation of the skeletal muscle program via MRFs in S. macrurus might differ from that of their mammalian counterparts. Together, these data further our understanding of the molecular processes involved in the plasticity of the vertebrate skeletal muscle program that brings about the muscle-like phenotype of the non-contractile electrogenic cells in S. macrurus.

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.

Serotonin modulates the electric waveform of the gymnotiform electric fish Brachyhypopomus pinnicaudatus.

The gymnotiform electric fish Brachyhypopomus pinnicaudatus communicates with a sexually dimorphic electric waveform, the electric organ discharge (EOD). Males display pronounced circadian rhythms in the amplitude and duration of their EODs. Changes in the social environment influence the magnitudes of these circadian rhythms and also produce more transient responses in the EOD waveforms. Here we show that injections of serotonin produce quick, transient, dose-dependent enhancements of the male EOD characters similar to those induced by encounters with another male. The response to serotonin administered peripherally begins 5-10 min post injection and lasts approximately 3 h. The magnitude of the response to serotonin is tightly associated with the magnitude of the day-to-night swing of the circadian rhythm prior to injection. Taken together these findings suggest that the male's social environment influences his response to serotonin by altering the function of some part of the downstream chain between the serotonin receptors and the ion channels involved in control of the EOD waveform. Although chronic activation of serotonin circuitry is widely known to elicit subordinate behavior, we find that 5-HT initially increases a dominance signal in these fish. These findings are consistent with the emerging view that serotonin facilitates different adaptive responses to acute and chronic social challenge and stress.