Sex-specific role of a glutamate receptor subtype in a pacemaker nucleus controlling electric behavior.

Electric communication signals, produced by South American electric fish, vary across sexes and species and present an ideal opportunity to examine the bases of signal diversity, and in particular, the mechanisms underlying sexually dimorphic behavior. Gymnotiforms produce electric organ discharges (EOD) controlled by a hindbrain pacemaker nucleus (PN). Background studies have identified the general cellular mechanisms that underlie the production of communication signals, EOD chirps and interruptions, typically displayed in courtship and agonistic contexts. Brachyhypopomus gauderio emit sexually dimorphic signals, and recent studies have shown that the PN acquires the capability of generating chirps seasonally, only in breeding males, by modifying its glutamatergic system. We hypothesized that sexual dimorphism was caused by sexual differences in the roles of glutamate receptors. To test this hypothesis, we analyzed NMDA and AMPA mediated responses in PN slice preparations by field potential recordings, and quantified one AMPA subunit mRNA, in the PNs of males and females during the breeding season. In situ hybridization of GluR2B showed no sexual differences in quantities between the male and female PN. Functional responses of the PN to glutamate and AMPA, on the other hand, showed a clear cut sexual dimorphism. In breeding males, but not females, the PN responded to glutamate and AMPA with bursting activity, with a temporal pattern that resembled the pattern of EOD chirps. In this study, we have been successful in identifying cellular mechanisms of sexual dimorphic communication signals. The involvement of AMPA receptors in PN activity is part of the tightly regulated changes that account for the increase in signal diversity during breeding in this species, necessary for a successful reproduction.

A central pacemaker that underlies the production of seasonal and sexually dimorphic social signals: anatomical and electrophysiological aspects.

Our long-term goal is to approach the understanding of the anatomical and physiological bases for communication signal diversity in gymnotiform fishes as a model for vertebrate motor pattern generation. Brachyhypopomus gauderio emits, in addition to its electric organ discharge (EOD) at basal rate, a rich repertoire of rate modulations. We examined the structure of the pacemaker nucleus, responsible for the EOD rate, to explore whether its high output signal diversity was correlated to complexity in its neural components or regional organization. We confirm the existence of only two neuron types and show that the previously reported dorsal-caudal segregation of these neurons is accompanied by rostral-caudal regionalization. Pacemaker cells are grouped dorsally in the rostral half of the nucleus, and relay cells are mainly ventral and more abundant in the caudal half. Relay cells are loosely distributed from the center to the periphery of the nucleus in correlation to somata size. Our findings support the hypothesis that regional organization enables a higher diversity of rate modulations, possibly offering distinct target areas to modulatory inputs. Since no anatomical or electrophysiological seasonal or sexual differences were found, we explored these aspects from a functional point of view in a companion article.

A central pacemaker that underlies the production of seasonal and sexually dimorphic social signals: functional aspects revealed by glutamate stimulation.

The cyclic enrichment of behavioral repertoires is a common event in seasonal breeders. Breeding males Brachyhypopomus gauderio produce electric organ discharge (EOD) rate modulations called chirps while females respond with interruptions. The electromotor system is commanded by a pacemaker nucleus (PN) which sets the basal rate and produces the rate modulations. We focused on identifying functional, seasonal and sexual differences in this nucleus in correlation to these differences in behavior. The in vivo response to glutamate injection in the PN was seasonal, sexually dimorphic and site specific. Non-breeding adults and breeding females injected in dorsal and ventral sites generated EOD rate increases and interruptions, respectively. Reproductive males added a conspicuous communication signal to this repertoire. They chirped repetitively when we injected glutamate in a very restricted area of the ventral-rostral nucleus, surprisingly one with a low number of relay cell somata. This study shows that the PN is functionally organized in regions in a caudal-rostral axis, besides the previously documented dorsal-ventral division. Functional regions are revealed by seasonal changes that annually provide this nucleus with the cellular mechanisms that allow the bursting activity underlying chirp production, only in males.

Vasotocin actions on electric behavior: interspecific, seasonal, and social context-dependent differences.

Social behavior diversity is correlated with distinctively distributed patterns of a conserved brain network, which depend on the action of neuroendocrine messengers that integrate extrinsic and intrinsic cues. Arginine vasotocin (AVT) is a key integrator underlying differences in behavior across vertebrate taxa. Weakly electric fish use their electric organ discharges (EODs) as social behavioral displays. We examined the effect of AVT on EOD rate in two species of Gymnotiformes with different social strategies: Gymnotus omarorum, territorial and highly aggressive, and Brachyhypopomus gauderio, gregarious and aggressive only between breeding males. AVT induced a long-lasting and progressive increase of EOD rate in isolated B. gauderio, partially blocked by the V1a AVT receptor antagonist (Manning compound, MC), and had no effects in G. omarorum. AVT also induced a long-lasting increase in the firing rate (prevented by MC) of the isolated medullary pacemaker nucleus (PN) of B. gauderio when tested in an in vitro preparation, indicating that the PN is the direct effector of AVT actions. AVT is involved in the seasonal, social context-dependent nocturnal increase of EOD rate that has been recently described in B. gauderio to play a role in mate selection. AVT produced the additional nocturnal increase of EOD rate in non-breeding males, whereas MC blocked it in breeding males. Also, AVT induced a larger EOD rate increase in reproductive dyads than in agonistic encounters. We demonstrated interspecific, seasonal, and context-dependent actions of AVT on the PN that contribute to the understanding of the mechanisms the brain uses to shape sociality.

Social electric signals in freely moving dyads of Brachyhypopomus pinnicaudatus.

Brachyhypopomus pinnicaudatus (pulse-type weakly electric fish) is a gregarious species that displays reproductive behavior and agonistic encounters between males only during the breeding season. During social interactions, in addition to its basal electric organ discharge (EOD), fish emit social electric signals (SESs) in the contexts of reproduction and intrasexual aggression. We reproduced natural behavior in laboratory settings: SESs recorded in the field are indistinguishable from those observed in our experimental setup. SESs are nocturnal, change seasonally and exhibit sexual dimorphism. This study provides an exhaustive characterization and classification of SESs produced by males and females during the breeding season. In male-female dyads, males produce accelerations and chirps while females interrupt their EODs. The same SESs are observed in male-male dyads. We present a novel, thorough classification of male chirps into four independent types (A, B, C, and M) based on their duration and internal structure. The type M chirp is only observed in male-male dyads. Chirps and interruptions, both in male-female and male-male dyads, are emitted in bouts, which are also grouped throughout the night. Our data suggest the existence of a sophisticated electric dialog during reproductive and aggressive interaction whose precise timing and behavioral significance are being investigated.

The effect of general and spinal anesthesia on balance control in elderly patients.

Falls are a major problem in the elderly population, but few communications address the influence of anesthesia on balance control. This study reports how a general balanced anesthesia (GBA) and a spinal anesthesia (SA) affect balance control in the elderly. We divided into three groups, according to electronystagmography findings and type of anesthesia, 21 men older than 65 years (mean age, 72 years) who were scheduled for prostate adenectomy. One group, designated GBN, consisted of normal subjects who underwent surgery under GBA. In another group, designated GBP, were pathological subjects who had clinically compensated central vestibular disorders (CVDs) and underwent surgery under GBA. The third group, designated SP, contained CVD patients who underwent surgery under SA. We assessed balance control via static posturography preoperatively and 48 hours postoperatively. We observed no change in balance control parameters (center of pressure distribution area [COPa] or COP sway velocity [SV]) for those patients in the GBN group or for those in the SP group. We did observe a significant difference for the patients in the GBP group, with higher postoperative values of COPa and SV (Wilcoxon signed rank test). Our results showed that in subjects with clinically compensated underlying CVD prior to a GBA, balance control worsens after the procedure, whereas no change in balance control occurs after an SA. Balance control in subjects with normal vestibuloocular function did not change even after a GBA.

Comunicación elcctrica en peces sudamericanos del orden gymnotiformes / Electric cmmuniccation in south american gymnotiform fishes

We present a description of the electrogenic system and the reproductive behavior of Gymnotiform electrical fishes focusing on sexual and seasonal variations of their electric displays, and the communicative value of their social electric signals roduced during courtship, using histological techniques and electrophysiological recordings. The pacemaker nucleus organizes the discharge sequence whereas the rest of the system determines the electric organ discharge waveform, which is species specific. During mating, Gymnotiform fishes show electric discharge sexual dimorphism, and generate transient social signals during courtship and gametoposition. Males of Brachyhypopomus pinnicaudatus prolong duration of the late phase of the discharge and generate brief signals characterized by rate increase and amplitude decrease called "Chirps". Females ransiently stop their discharge. These courtship transient signals are always produced over a specially large nocturnal rate increase. Chirps and interruptions are repeated in bursts with a temporal association that suggests communication.

Se presenta la organización del sistema electrogénico y de la conducta reproductiva de los peces eléctricos Gymnnotiformes enfatizando las variaciones sexuales y estacionales de sus despliegues eléctricos mediante histología y registros electrofisiológicos de la forma de onda de la descarga y de la actividad de las células componentes. Identificamos un núcleo electro-motor bulbar que activa la descarga eléctrica del pez. En la reproducción, los Gymnotiformes exhiben dimorfismo sexual de la frecuencia de descarga y producen señales para el cortejo y la gametoposición. Los machos de Brachyhypopomus pinnicaudatus exhiben prolongación de la segunda fase de la Descarga del Órgano Eléctrico (DOE) y emiten señales breves con aumento brusco de la frecuencia y disminución de la amplitud (Chirps). Las hembras interrumpen su descarga de forma transitoria. Estas señales se sobreimponen a un aumento nocturno de la frecuencia basal. Chirps e interrupciones se producen en ráfagas con asociación temporal precisa que sugiere comunicación.

Environmental, seasonal, and social modulations of basal activity in a weakly electric fish.

The electric organ discharge (EOD) of weakly electric fish encodes information about species, sex, behavioral, and physiological states throughout the lifetime. Its central command is crucial for sensory-motor coordination, and is therefore the target of plastic mechanisms that adapt fish to environmental and social challenges. The EOD waveform of Brachyhypopomus pinnicaudatus is modulated by environmental factors and the neuroendocrine system. In this study we investigate the effects of water temperature and day-night cycle upon EOD rate in this species during the breeding and non-breeding seasons. During the non-breeding season, EOD rate is a linear function of water temperature and exhibits counterclockwise hysteresis. During breeding, a thermal resistance strategy prevents the decrease of EOD rate to cooling. A nocturnal increase of EOD basal rate independent of water temperature and locomotor activity was demonstrated in isolated non-breeding adults and in male-female dyads all year round. An additional increase of nocturnal EOD rate, probably acting as a social courtship signal, was found in breeding dyads. This additional increase of nocturnal EOD rate could not be fully explained by gonadal maturation and was modulated by social stimuli. This study provides novel data on the complex interactions between environment, reproductive cycle, social behavior, and electromotor output in an advantageous model of the vertebrate central nervous system.

Influence of temperature and reproductive state upon the jamming avoidance response in the pulse-type electric fish Brachyhypopomus pinnicaudatus.

The electric organ discharge (EOD) in Brachyhypopomus pinnicaudatus is modified by temperature and reproductive state. We studied the influence of these variables upon a complex behavior, the jamming avoidance response (JAR). Experiments were performed in non-reproductive fish and in two groups of fish after the induction of reproductive state (by nature or by acclimation at 28 degrees C). JARs were elicited at 20 and 30 degrees C by free-run electric stimuli with different deltaLs (interval difference between stimulus and EOD). In non-reproductive fish, JARs induced by stimuli with +deltaLs showed temperature sensitivity, with smaller responses at 30 degrees C. Conversely, similar JARs were obtained at both temperatures in reproductive fish. These observations were replicated in curarized preparations. Stimuli with -deltaLs were almost ineffective in non-reproductive fish at 30 degrees C, whereas adequate JARs were shown by reproductive fish. Phase-locked stimuli were used to evaluate the duration of the low-threshold electrosensory periods preceding and following the EOD. In non-reproductive fish, the temperature step induced a shortening of these periods. The opposite effect was observed in reproductive fish, probably explaining the differences in JAR capability. A prolongation of the low-threshold periods would favor the perception of electrocommunication signals during courtship. JAR changes would be a consequence of this adaptation.

Sodium-dependent plateau potentials in electrocytes of the electric fish Gymnotus carapo.

The weakly electric fish Gymnotus carapo emits a triphasic electric organ discharge generated by muscle-derived electrocytes, which is modified by environmental and physiological factors. Two electrode current clamp recordings in an in vitro preparation showed that Gymnotus electrocytes fired repetitively and responded with plateau potentials when depolarized. This electrophysiological behavior has never been observed in electrocytes from related species. Two types of plateaus with different thresholds and amplitudes were evoked by depolarization when Na(+)-dependent currents were isolated in a K(+)- and Ca(2+)-free solution containing TEA and 4-AP. Two electrode voltage clamp recordings revealed a classical fast activating-inactivating Na+ current and two persistent Na(+)-dependent currents with voltage-dependencies consistent with the action potential (AP) and the two plateaus observed under current clamp, respectively. The three currents, the APs and the plateaus were reduced by TTX, and were absent in Na(+)-free solution. The different Na(+)-dependent currents in Gymnotus electrocytes may be targets for the modifications of the electric organ discharge mediated by environmental and physiological factors.

Temperature induces gonadal maturation and affects electrophysiological sexual maturity indicators in Brachyhypopomus pinnicaudatus from a temperate climate.

In contrast to most of the previous studies in gymnotiform reproduction, which have been conducted in the tropical region, this study examines a gymnotid from the temperate region in both the natural habitat and the laboratory. The gonadal histology of Brachyhypopomus pinnicaudatus is described for the first time. The male had a paired, lobular testis of the unrestricted spermatogonial type, and females a paired saccular cystovary. Analysis of gonads and their annual cycle enabled us to confirm the breeding season and to conclude that this species is a multiple spawner. Water temperature and photoperiod showed the expected annual cycles for the region. High temperature and a 14 h:10 h L:D photoperiod in the natural habitat coincided with (1) mature gonadal stages, (2) electrophysiological sexual dimorphism: males present a lengthened negative phase in their electric organ discharge (EOD) and (3) decreased temperature sensitivity of the EOD: the waveform does not change when temperature increases above 20 degrees C. Acclimation to sustained high temperature (30 days, 28 degrees C, 12 h:12 h L:D, low conductivity) induced gonad maturation along with EOD dimorphism. Our data show that high environmental temperature is enough to trigger sexual maturity in Brachyhypopomus pinnicaudatus from a temperate climate.

Environmental and hormonal influences upon EOD waveform in gymnotiform pulse fish.

Temperature is a major variable that affects all biological systems. Environmental temperature determines animal geographical distribution and activity, and influences their reproductive cycle, particularly within the temperate zone. Temperature, as a physical parameter, also strongly affects excitable tissues. The hypothesis of temperature as the most important environmental cue for the onset of breeding in gymnotiform pulse fish of the temperate zone is supported by: (a) a clear temporal correlation that was observed in the wild between water temperature and sexual maturity, and (b) the induction of gonadal maturation and sexual differences after acclimation at high temperature (28 degrees C) in the laboratory. Temperature sensitivity of EOD waveform (described in Brachyhypopomus pinnicaudatus and Gymnotus carapo) is characterized by the decrease of the EOD's late head-negative phase as temperature increases. This phenomenon depends on electrocyte properties since: (a) experimentally induced changes of discharge rate at constant temperature generate smaller EOD distortion, and (b) the effect of temperature upon EOD also depends on water conductivity. Temperature sensitivity of EOD waveform is negatively correlated with gonadal maturity in Brachyhypopomus pinnicaudatus. High temperature sensitivity was observed during the non-breeding season, whereas low temperature sensitivity was recorded during the breeding season. Temperature sensitivity of EOD waveform in both Brachyhypopomus pinnicaudatus and Gymnotus carapo was modulated by: (a) testosterone treatment (100 microg/g) that decreased temperature sensitivity, and (b) acclimation at high temperature (28 degrees C, 1 month) that also decreased temperature sensitivity. Temperature is probably acting through the neuroendocrine system, and ultimately interacting with steroid hormones in their effects upon EOD waveform.