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.

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.

Neural substrate of an increase in sensory sampling triggered by a motor command in a gymnotid fish.

Despite recent advances that have elucidated the effects of collateral of motor commands on sensory processing structures, the neural mechanisms underlying the modulation of active sensory systems by internal motor-derived signals remains poorly understood. This study deals with the neural basis of the modulation of the motor component of an active sensory system triggered by a central motor command in a gymnotid fish. In Gymnotus omarorum, activation of Mauthner cells, a pair of reticulospinal neurons responsible for the initiation of escape responses in most teleosts, evokes an abrupt and prolonged increase in the rate of the electric organ discharge (EOD), the output signal of the electrogenic component of the active electrosensory system. We show here that prepacemaker neural structures (PPs) that control the discharge of the command nucleus for EODs are key elements of this modulation. Retrograde labeling combined with injections of glutamate at structures that contain labeled neurons showed that PPs are composed of a bilateral group of dispersed brain stem neurons that extend from the diencephalon to the caudal medulla. Blockade of discrete PPs regions during the Mauthner cell-initiated electrosensory modulation indicate that the long duration of this modulation relied on activation of diencephalic PPs, whereas its peak amplitude depended on the recruitment of medullary PPs. Temporal correlation of motor and sensory consequences of Mauthner cell activation suggests that the Mauthner cell-initiated enhancement of electrosensory sampling is involved in the selection of escape trajectory.

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.

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.

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.