Pharmacological study of the one spike spherical neuron phenotype in Gymnotus omarorum.

The intrinsic properties of spherical neurons play a fundamental role in the sensory processing of self-generated signals along a fast electrosensory pathway in electric fish. Previous results indicate that the spherical neuron's intrinsic properties depend mainly on the presence of two resonant currents that tend to clamp the voltage near the resting potential. Here we show that these are: a low-threshold potassium current blocked by 4-aminopyridine and a mixed cationic current blocked by cesium chloride. We also show that the low-threshold potassium current also causes the long refractory period, explaining the necessary properties that implement the dynamic filtering of the self-generated signals previously described. Comparative data from other fish and from the auditory system indicate that other single spiking onset neurons might differ in the channel repertoire observed in the spherical neurons of Gymnotus omarorum.

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.

Peripheral electrosensory imaging by weakly electric fish.

Different species have developed different solutions to the problem of constructing a representation of the environment from sensory images projected onto sensory surfaces. Comprehension of how these images are formed is an essential first step in understanding the representation of external reality by a given sensory system. Modeling of the electrical sensory images of objects began with the discovery of electroreception and continues to provide general insights into the mechanisms of imaging. Progress in electric image research has made it possible to establish the physical basis of electric imaging, as well as methods to accurately predict the electric images of objects alone and as a part of a natural electric scene. In this review, we show the following. (1) The internal low resistance of the fish's body shapes the image in two different ways: by funneling the current generated by the electric organ to the sensory surface, it increases the fields rostrally, thus enhancing the perturbation produced by nearby objects; and by increasing the projected image. (2) The electric fish's self-generated currents are modified by capacitive objects in a distinctive manner. These modulations can be detected by different receptor types, yielding the possibility of "electric color." (3) The effects of different objects in a scene interact with each other, generating an image that is different from the simple addition of the images of individual objects, thus causing strong contextual effects.

The electric image in Gnathonemus petersii.

We review modelling and experimental work dealing with the mechanisms of generation of electric image. We discuss: (1) the concept of electric image in the context of the reafference principle; (2) how waveform codes an impedance related qualia of the object image, referred to as "electric colour"; (3) that some characteristics of the spatial profiles generated by pre-receptor mechanisms are suitable for edge detection; (4) which parameters of the spatial profiles provide information for distance discrimination; (5) that electric images are distributed representations of the scene.

Electroreception in Gymnotus carapo: differences between self-generated and conspecific-generated signal carriers.

Local electric fields generated by the electric organ discharge of Gymnotus carapo were explored at selected points on the skin of an emitter fish ('local self-generated fields') and on the skin of a conspecific ('local conspecific-generated fields') using a specially designed probe. Local self-generated fields showed a constant pattern along the body of the fish. At the head, these fields were collimated, much stronger than elsewhere on the fish, and had a time waveform that was site-independent. This waveform consisted of a slow head-negative wave followed by a faster head-positive wave. In contrast, time waveforms in the trunk and tail regions were site-specific, with field vectors that changed direction over time. Local conspecific-generated fields were similar to the head-to-tail field, but their spatio-temporal pattern at the skin depended on the relative orientation between the receiving fish and the emitting fish. Because self-generated fields had a slow early component at the head region, they displayed a low-frequency peak in their power spectral density histograms. In contrast, the conspecific-generated fields had time waveforms with a sharper phase reversal, resulting in a peak at higher frequency than in the self-generated field. Lesions in emitting fish demonstrated that waveform components generated by the trunk and tail regions of the electric organ predominate in conspecific-generated fields, whereas waveform components generated by the abdominal region prevail in self-generated fields. Similar results were obtained from Brachyhypopomus pinnicaudatus. These results suggest that, in pulse-emitting gymnotids, electrolocation and electrocommunication signals may be carried by different field components generated by different regions of the electric organ.

Electroreception in Gymnotus carapo: pre-receptor processing and the distribution of electroreceptor types.

This paper describes the peripheral mechanisms involved in signal processing of self- and conspecific-generated electric fields by the electric fish Gymnotus carapo. The distribution of the different types of tuberous electroreceptor and the occurrence of particular electric field patterns close to the body of the fish were studied. The density of tuberous electroreceptors was found to be maximal on the jaw (foveal region) and very high on the dorsal region of the snout (parafoveal region), decaying caudally. Tuberous type II electroreceptors were much more abundant than type I electroreceptors. Type I electroreceptors occurred exclusively on the head and rostral trunk regions, while type II electroreceptors were found along as much as 90 % of the fish. Electrophysiological data indicated that conspecific- and self-generated electric currents are 'funnelled' by the high conductivity and geometry of the body of the fish. These currents are concentrated at the peri-oral zone, where most electroreceptors are located. Moreover, within this region, field vector directions were collimated, constituting the most efficient stimulus for electroreceptors. It can be concluded that the passive properties of the fish tissue represent a pre-receptor device that enhances exafferent and reafferent electrical signals at the fovea-parafoveal region.

The electric image in weakly electric fish: perception of objects of complex impedance.

Weakly electric fish explore the environment using electrolocation. They produce an electric field that is detected by cutaneous electroreceptors; external objects distort the field, thus generating an electric image. The electric image of objects of complex impedance was investigated using a realistic model, which was able to reproduce previous experimental data. The transcutaneous voltage in the presence of an elementary object is modulated in amplitude and waveform on the skin. Amplitude modulation (measured as the relative change in the local peak-to-peak amplitude) consists of a 'Mexican hat' profile whose maximum relative slope depends on the distance of the fish from the object. Waveform modulation depends on both the distance and the electrical characteristics of the object. Changes in waveform are indicated by the amplitude ratio of the larger positive and negative phases of the local electric organ discharge on the skin. Using the peak-to-peak amplitude and the positive-to-negative amplitude ratio of this discharge, a perceptual space can be defined and correlated with the capacitance and resistance of the object. When the object is moved away, the perceptual space is reduced but keeps the same proportions (homothetically): for a given object, the positive-to-negative amplitude ratio is a linear function of the peak-to-peak amplitude. This linear function depends on the electrical characteristics of the object. However, there are 'families' of objects with different electrical characteristics that produce changes in the parameters of the local electric organ discharge that are related by the same linear function. We propose that these functions code the perceptual properties of an object related to its impedance.

Structural and functional aspects of the fast electrosensory pathway in the electrosensory lateral line lobe of the pulse fish Gymnotus carapo.

The fast electrosensory pathway (FEP) of gymnotiform fish is mediated by tuberous electroreceptor organs innervated by ganglion cells that synapse with spherical cells of the electrosensory lateral line lobe (ELL). Spherical cells project to the magnocellular mesencephalic nucleus. The electrosensory environment was represented somatotopically within ELL. The mandibular (MN) and the supraorbital (SON) nerves projected to rostral ELL (occupying 19-28% and 4-10%, respectively), and the posterior branch of the anterior lateral line nerve (pALLN) projected to caudal ELL (occupying 56-64%). Labeling with horseradish peroxidase or biotinylated dextran-amine demonstrated three kinds of synaptic endings coupling primary afferents to spherical cells: multiple synaptic knobs, medium-sized calyxes, and very large calyxes. Multiple synaptic knobs arose from MN and SON primary afferents and were found in a narrow rostral area covering the centrolateral (CLS) and lateral (LS) segments of ELL. Medium and large calyxes, proceeding from the same nerves, predominated in the remaining parts of the three segments of ELL containing spherical cells. Calyx-type endings were also found in the LS-occupying regions in which the pALLNs projected. Calyx-type endings formed gap junctions but also contained vesicles and showed submembrane specializations typical of chemical synapses. The postsynaptic spherical cells were linked by dendrosomatic gap junctions and were also contacted by unlabeled en passant synaptic boutons, whose fine structure suggested chemical transmission. Electrophysiological studies indicated that spherical cell responsiveness diminished after electrosensory stimulation. This apparently inhibitory phenomenon may be subserved by the unlabeled synaptic boutons, which possibly originate from interneurons that have yet to be identified.

The electric organ discharge of Brachyhypopomus pinnicaudatus. The effects of environmental variables on waveform generation.

The electric organ discharge of Brachyhypopomus pinnicaudatus was studied by recording (1) the discharge field potentials in water at different conductivities and temperatures and (2) the spatiotemporal pattern of electromotive forces of the equivalent source. An early deflection, head positive (P wave), and a late deflection, head negative (N wave), are the major components of the discharge, however a striking double positive peak is generated at the abdominal level. Comparisons of this species with other pulse gymnotids provide evidence for common patterns of organization of the electrogenic system: (1) There is a head-to-tail activation wave along the fish; (2) the electromotive force increases exponentially from head to tail, but it is differentially attenuated by the passive tissues in male and females; (3) the abdominal region generates a complex species-specific waveform, whereas the tail discharge is similar across species. In B. pinnicaudatus the electric organ discharge waveform is sensitive to endocrine and environmental stimuli. The effect of seasonal sex differences on electrogenic and passive tissue, the changes in impedance matching between the fish's body and the environment, and the modulation of membrane properties by temperature, are able to modify the EOD waveform. Since these factors change during the breeding season, their appropriate combination might be crucial for reproduction.

The electric image in weakly electric fish: physical images of resistive objects in Gnathonemus petersii.

The present study describes a measurement-based model of electric image generation in the weakly electric mormyrid fish Gnathonemus petersii. Measurements of skin impedance, internal resistivity and fish body dimensions have been used to generate an electrical-equivalent model of the fish and to calculate electrical images and equivalent dipole sources for elementary resistive objects. These calculations allow us to understand how exafferent and reafferent signals are sensed by electroreceptors. An object's electric image consists of the modulation of the transcutaneous voltage profile generated by the fish's own discharge. The results suggest a set of rules for electrolocation: (1) the side of the fish where modulation is larger indicates the side on which the object is situated; (2) the object's position in the electroreceptive field is indicated by the point of maximum modulation of the transcutaneous voltage; (3) the degree of focus of the image indicates the distance to the object. In addition, center-surround opposition originating at pre-receptor level is proposed. Both experimental measurements and modeling indicate that fish skin impedance is relatively low (400-11 000 cm²) and mainly resistive. This low skin impedance appears to enhance the local electric organ discharge modulation, the center-surround effect, the signal-to-noise ratio for electrolocation and the active space for electrocommunication.

The electric image in weakly electric fish: I. A data-based model of waveform generation in Gymnotus carapo.

Understanding how electrosensory images are generated and perceived in actively electrolocating fish requires the study of the characteristics of fish bodies as electric sources. This paper presents a model of Gymnotus carapo based on measurements of the electromotive force generated by the electric organ and the impedance of the passive tissues. A good agreement between simulated and experimentally recorded transcutaneous currents was obtained. Passive structures participate in the transformation of the electromotive force pattern into transcutaneous current profiles. These spatial filtering properties of the fish's body were investigated using the model. The shape of the transcutaneous current profiles depends on tissue resistance and on the geometry and size of the fish. Skin impedance was mainly resistive. The effect of skin resistance on the spatial filtering properties of the fish's body was theoretically analyzed. The model results show that generators in the abdominal and central regions produce most of the currents through the head. This suggests that the electric organ discharge (EOD), generated in the abdominal and central regions is critical for active electrolocation. In addition, the well-synchronized EOD components generated all along the fish produce large potentials in the far field. These components are probably involved in long-distance electrocommunication. Preliminary results of this work were published as a symposium abstract.

The spinal cord of Gymnotus carapo: the electromotoneurons and their projection patterns.

The electric organ of Gymnotus carapo lies parallel to the spinal cord and extends from the pectoral girdle to the tip of the tail. The spinal electromotoneurons are distributed in a relatively consistent pattern: there is a peak at 17% of the fish's length and an irregular distribution beyond 25%. Horseradish peroxidase injections into the electric organ not exceeding 5% of the fish's length labeled electromotoneuron arrays occupying 20% of the fish's length. Injections made in four discrete rostrocaudal electric organ regions resulted in labeled electromotoneurons distributed along four sequential but overlapping arrays. Since the caudal portion of the spinal cord lacks electromotoneurons, there is a shortened representation of the electric organ. The electromotoneuron population is not homogeneous: there are small neurons (somata 25-40 microns) and large neurons (somata 45-60 microns) unevenly distributed along the cord. Small neurons occur at more rostral spinal cord segments, while large neurons lie in more caudal segments. Both kinds of nerve cells coexist in the intermediate regions. Overlapping of subsequent neuronal arrays favors synchronized firing of electrocytes. The presence of two neuronal populations differing in size and projecting to opposite electrocyte faces may account for the timed excitation of the electrogenic surfaces. Taking into account these new findings a comprehensive explanation of the activation sequence along the spinal cord and the electric organ is proposed.

Velocity of ultrasound in active and passive cat medial gastrocnemius muscle.

The lengths and pinnation angles of muscle fibers in the medial gastrocnemius (MG) muscle have recently been measured in freely moving cats [Hoffer et al., Progr. Brain Res. 80, 75-85 (1989); Muscle Afferents and Spinal Control of Movement (1992)] using an ultrasound transit-time (USTT) technique. This method assumed that the velocity of ultrasound through intact muscles was constant, independent of fiber orientation, muscle activity, load, belly shape, or fiber movement. However, the velocity of ultrasound along and across the fibers has been reported to depend on the state of muscle activation in frog muscle experiments in vitro [Hatta et al., J. Physiol. 403, 193-209 (1988)]. In the present study, the assumption of constant velocity of ultrasound in the cat MG muscle was evaluated. In acute experiments, done in situ with intact blood supply, the USTT was measured along and across cat MG muscle fibers in the passive, reflexly activated and tetanically activated states, with and without changes in muscle fiber length, for situations that reproduced the length and force ranges normally used by cats during locomotion. The velocity of ultrasound was found to be independent of the state of activation or motion of the muscle, and independent of the direction of the measurement with respect to the fiber orientation, within a measurement uncertainty less than or equal to 0.2%. These results validate the use of the USTT technique for the measurement of intramuscular dimensions in freely moving animals.