Apteronotus Leptorhynchus Research Articles

Nonrenewal Statistics of Electrosensory Afferent Spike Trains: Implications for the Detection of Weak Sensory Signals

The ability of an animal to detect weak sensory signals is limited, in part, by statistical fluctuations in the spike activity of sensory afferent nerve fibers. In weakly electric fish, probability coding (P-type) electrosensory afferents encode amplitude modulations of the fish's self-generated electric field and provide information necessary for electrolocation. This study characterizes the statistical properties of baseline spike activity in P-type afferents of the brown ghost knifefish, Apteronotus leptorhynchus. Short-term variability, as measured by the interspike interval (ISI) distribution, is moderately high with a mean ISI coefficient of variation of 44%. Analysis of spike train variability on longer time scales, however, reveals a remarkable degree of regularity. The regularizing effect is maximal for time scales on the order of a few hundred milliseconds, which matches functionally relevant time scales for natural behaviors such as prey detection. Using high-order interval analysis, count analysis, and Markov-order analysis we demonstrate that the observed regularization is associated with memory effects in the ISI sequence which arise from an underlying nonrenewal process. In most cases, a Markov process of at least fourth-order was required to adequately describe the dependencies. Using an ideal observer paradigm, we illustrate how regularization of the spike train can significantly improve detection performance for weak signals. This study emphasizes the importance of characterizing spike train variability on multiple time scales, particularly when considering limits on the detectability of weak sensory signals.

Spontaneous modulations of the electric organ discharge in the weakly electric fish, Apteronotus leptorhynchus: a biophysical and behavioral analysis.

Brown ghosts, Apteronotus leptorhynchus, are weakly electric gymnotiform fish whose wave-like electric organ discharges are distinguished by their enormous degree of regularity. Despite this constancy, two major types of transient electric organ discharge modulations occur: gradual frequency rises, which are characterized by a relatively fast increase in electric organ discharge frequency and a slow return to baseline frequency; and chirps, brief and complex frequency and amplitude modulations. Although in spontaneously generated gradual frequency rises both duration and amount of the frequency increase are highly variable, no distinct subtypes appear to exist. This contrasts with spontaneously generated chirps which could be divided into four "natural" subtypes based on duration, amount of frequency increase and amplitude reduction, and time-course of the frequency change. Under non-evoked conditions, gradual frequency rises and chirps occur rather rarely. External stimulation with an electrical sine wave mimicking the electric field of a neighboring fish leads to a dramatic increase in the rate of chirping not only during the 30 s of stimulation, but also in the period immediately following the stimulation. The rate of occurrence of gradual frequency rises is, however, unaffected by such a stimulation regime.

Temperature Dependence of Electrocommunication Signals and Their Underlying Neural Rhythms in the Weakly Electric Fish, Apteronotus leptorhynchus

Weakly electric fish emit an electric communication signal that is controlled by a highly specialized neural circuit. In Apteronotus, the continuous electric organ discharge (EOD) is generated by electrotonically coupled neurons in the hindbrain pacemaker nucleus, and transient EOD modulations involve chemical synapses from descending midbrain and thalamic prepacemaker nuclei. We characterized the effects of temperature change (18–32°C) on both the continuous EOD and EOD modulations, chirps, in A. leptorhynchus. EOD frequency was linearly related to temperature (Q₁₀=1.62). By contrast, the temperature dependence of EOD amplitude changed with temperature. Amplitude increased steeply with temperature below 25°C (Q₁₀=2.0), but increased only gradually above 25°C (Q₁₀=1.15). EOD waveform, and consequently harmonic content, was also affected by temperature. The amplitude of the second harmonic was relatively high at both low and high temperature and relatively low at intermediate temperatures. The amplitude of the third harmonic increased monotonically with temperature. Thus, temperature has qualitative as well as quantitative effects on the production of the EOD. Chirp rate (Q₁0=3.2) had a higher temperature dependence than that of the continuous EOD, which likely reflects its reliance on chemical rather than electrotonic synapses. In vitro pacemaker firing frequency had a similar, but slightly higher Q₁₀ (1.82) than that of the EOD frequency.

Pharmacological characterization of ionic currents that regulate the pacemaker rhythm in a weakly electric fish.

Electric organ discharge (EOD) frequency in the brown ghost knifefish (Apteronotus leptorhynchus) is sexually dimorphic, steroid-regulated, and determined by the discharge rates of neurons in the medullary pacemaker nucleus (Pn). We pharmacologically characterized ionic currents that regulate the firing frequency of Pn neurons to determine which currents contribute to spontaneous oscillations of these neurons and to identify putative targets of steroid action in regulating sexually dimorphic EOD frequency. Tetrodotoxin (TTX) initially reduced spike frequency, and then reduced spike amplitude and stopped pacemaker activity. The sodium channel blocker muO-conotoxin MrVIA also reduced spike frequency, but did not affect spike amplitude or production. Two potassium channel blockers, 4-aminopyridine (4AP) and kappaA-conotoxin SIVA, increased pacemaker firing rates by approximately 20% and then stopped pacemaker firing. Other potassium channel blockers (tetraethylammonium, cesium, alpha-dendrotoxin, and agitoxin-2) did not affect the pacemaker rhythm. The nonspecific calcium channel blockers nickel and cadmium reduced pacemaker firing rates by approximately 15-20%. Specific blockers of L-, N-, P-, and Q-type calcium currents, however, were ineffective. These results indicate that at least three ionic currents-a TTX- and muO-conotoxin MrVIA-sensitive sodium current; a 4AP- and kappaA-conotoxin SIVA-sensitive potassium current; and a T- or R-type calcium current-contribute to the pacemaker rhythm. The pharmacological profiles of these currents are similar to those of currents that are known to regulate firing rates in other spontaneously oscillating neural circuits.

Precision of the pacemaker nucleus in a weakly electric fish: network versus cellular influences.

We investigated the relative influence of cellular and network properties on the extreme spike timing precision observed in the medullary pacemaker nucleus (Pn) of the weakly electric fish Apteronotus leptorhynchus. Of all known biological rhythms, the electric organ discharge of this and related species is the most temporally precise, with a coefficient of variation (CV = standard deviation/mean period) of 2 x 10(-4) and standard deviation (SD) of 0.12-1.0 micros. The timing of the electric organ discharge is commanded by neurons of the Pn, individual cells of which we show in an in vitro preparation to have only a slightly lesser degree of precision. Among the 100-150 Pn neurons, dye injection into a pacemaker cell resulted in dye coupling in one to five other pacemaker cells and one to three relay cells, consistent with previous results. Relay cell fills, however, showed profuse dendrites and contacts never seen before: relay cell dendrites dye-coupled to one to seven pacemaker and one to seven relay cells. Moderate (0.1-10 nA) intracellular current injection had no effect on a neuron's spiking period, and only slightly modulated its spike amplitude, but could reset the spike phase. In contrast, massive hyperpolarizing current injections (15-25 nA) could force the cell to skip spikes. The relative timing of subthreshold and full spikes suggested that at least some pacemaker cells are likely to be intrinsic oscillators. The relative amplitudes of the subthreshold and full spikes gave a lower bound to the gap junctional coupling coefficient of 0.01-0.08. Three drugs, called gap junction blockers for their mode of action in other preparations, caused immediate and substantial reduction in frequency, altered the phase lag between pairs of neurons, and later caused the spike amplitude to drop, without altering the spike timing precision. Thus we conclude that the high precision of the normal Pn rhythm does not require maximal gap junction conductances between neurons that have ordinary cellular precision. Rather, the spiking precision can be explained as an intrinsic cellular property while the gap junctions act to frequency- and phase-lock the network oscillations.

Differential expression of the PSD-95 gene family in electrosensory neurons.

The PSD-95 family of membrane-associated guanylate kinase (MAGUK) proteins are involved in the assembly and organization of neurotransmitter receptors at excitatory synapses in the vertebrate nervous system. We have isolated partial cDNAs for five PSD-95 family members from Apteronotus leptorhynchus brain RNA using a degenerate PCR method. The amino acid sequences deduced indicate that A. leptorhynchus neurons express homologues of the mammalian PSD-93, SAP-97, and SAP-102 MAGUKs and two homologues of mammalian PSD-95. In situ hybridization experiments have been carried out to localize the cellular expression of all five MAGUK mRNAs in the central nervous system of A. leptorhynchus. In the cerebellum the expression patterns are highly similar to patterns reported for mammalian cerebellum, suggesting an evolutionary conservation of the functional roles in this gene family. Cellular levels of expression of the PSD-95 MAGUK mRNAs and the NMDAR-1 mRNA were highly correlated in neurons of the dorsal forebrain but were not correlated in neurons of the electrosensory lateral line lobe (ELL) or the cerebellum. These results suggest that the expression of PSD-95 MAGUK genes in forebrain neurons may provide mechanisms for synaptic organization that are not shared by neurons in the ELL and cerebellum.

Cell Proliferation after Lesions in the Cerebellum of Adult Teleost Fish: Time Course, Origin, and Type of New Cells Produced

In contrast to mammals, fish exhibit an enormous capacity to replace damaged neurons following injuries to the adult central nervous system. As the mechanisms controlling this so-called neuronal regeneration are unknown, we have, in the present study, examined the role of cell proliferation in this process. Lesions were applied to one subdivision of the cerebellum, the corpus cerebelli, in the teleost fish Apteronotus leptorhynchus. Proliferative activity was monitored through incorporation of the thymidine analogue 5-bromo-2′-deoxyuridine into replicating DNA. Cerebellar lesions induce high proliferative activity especially in areas in close vicinity to the injury, although the number of cells produced is also increased in other regions of the corpus cerebelli. Many of the cells generated in these areas become, after migration, specifically incorporated at the site of the lesion. The vast majority of them is dividing between 1 and 10 days following the lesion, with the maximum proliferative activity occurring at 5 days. Remarkably, also cells dividing 2 days prior to applying a lesion participate, at a significant number, in the regenerative process. Combination of 5-bromo-2′-deoxyuridine labeling with retrograde tract-tracing techniques demonstrated that at least some of the new cells that replace damaged neurons are cerebellar granule cells. This ability to generate new neurons, together with the previously described occurrence of apoptosis to remove damaged cells, is likely to form the basis for the enormous capacity of teleost fish to perform neuronal regeneration.

Ionic currents that contribute to a sexually dimorphic communication signal in weakly electric fish.

Weakly electric fish produce a communication signal, the electric organ discharge, that is species specific, and in many species, sexually dimorphic. Because the neural circuit that controls the electric organ discharge is relatively simple, it is an excellent model in which to study both the biophysical mechanisms underlying a rhythmic behavior and the neuroendocrine control of a sexually dimorphic behavior. By studying the effects of ion channel blockers on neurons in the medullary pacemaker nucleus, I pharmacologically characterized three ionic currents that influence the pacemaker rhythm, and thus electric organ discharge frequency, in the gymnotiform fish, Apteronotus leptorhynchus. These currents included a tetrodotoxin-sensitive sodium current; a potassium current that was sensitive to 4-aminopyridine; and a calcium current that was sensitive to nickel and cadmium, but resistant to specific blockers of L-, N-, P-, and Q-type calcium currents. The pharmacological profiles of the ionic currents in the pacemaker nucleus are similar to those of ionic currents involved in pacemaking in other neuronal oscillators. Because these ionic currents dramatically influence pacemaker firing frequency, which is directly related to electric organ discharge frequency, these ionic currents are likely targets of steroid hormone action in producing sexual dimorphisms in electric organ discharge frequency. Additional studies are needed to determine how these ionic currents interact to generate the electric organ discharge rhythm and to investigate the possibility that sexual dimorphism in the electric organ discharge results from the actions of gonadal steroids on these ionic currents.

Up-regulation of somatostatin after lesions in the cerebellum of the teleost fish Apteronotus leptorhynchus

Following application of mechanical lesions to the corpus cerebelli, a cerebellar subdivision, in adult individuals of the teleost fish Apteronotus leptorhynchus, the pattern of expression of the neuropeptide somatostatin was examined by employing immunohistochemical techniques. In the intact corpus cerebelli, only a very few cells displayed somatostatin-like immunoreactivity. This number dramatically increased in the area of the lesion within the granule cell layer 1 day following the injury and peaked after 2 days, when the normalized total number of somatostatin-positive cells was approximately 50 times higher than the mean number of labeled cells found after 3, 6, and 12 h of survival. Between 5 and 10 days of post-lesioning survival time, this number abruptly declined and returned to background levels at 17 and 25 days. Confocal microscopy revealed three cell types, presumably corresponding to granule cell neurons, astrocytes and microglia, which all displayed a similar temporal pattern of somatostatin expression. It is hypothesized that somatostatin is involved in regulation of the genesis and/or development of new neurons which are produced in response to injuries and which replace damaged cells at the site of the lesion.

Distribution of calcium/calmodulin-dependent kinase 2 in the brain of Apteronotus leptorhynchus.

Antibodies directed against the mammalian alpha and beta subunits of calcium/calmodulin-dependent kinase 2 (CaMK2) and brain dissection were used for immunoblot analysis of these proteins in various brain regions of Apteronotus leptorhynchus. Western blots revealed that the CaMK2alpha antibody labeled a single band of the expected molecular mass (approximately 50 kDa) for this enzyme in rat cortex and electric fish brain. CaMK2alpha was enriched in fish forebrain and hypothalamus and also strongly expressed in midbrain sensory areas. Western blots revealed that CaMK2beta antibodies labeled bands in an appropriate molecular mass range (approximately 58-64 kDa) for this enzyme in mammalian cortex and electric fish brain. However, a higher molecular mass band (approximately 80 kDa) was also labeled; because all these bands were eliminated by preadsorbtion with the CaMK2-derived peptide antigen, they may all represent CaMK2beta-like isoforms. We mapped the brain distribution of CaMK2 isoforms with emphasis on the electrosensory system. CaMK2alpha was present at high density in dorsal forebrain, hypothalamic nuclei, torus semicircularis, and tectum. It was also enriched in discrete fiber tracts in forebrain, diencephalon, and rhombencephalon. CaMK2beta-like isoforms were enriched in ventral forebrain, hypothalamic nuclei, torus semicircularis and the reticular formation. Unlike CaMK2alpha, CaMK2beta -like isoforms were predominantly present in cell bodies and rarely found in fiber tracts or neuropil. In the electrosensory lateral line lobe, CaMK2alpha was restricted to specific feedback fibers, i.e., tractus stratum fibrosum and its terminal field in the ventral molecular layer. In contrast, CaMK2beta-like isoforms were enriched in somata and dendrites of pyramidal cells and granular interneurons.

Neurogenesis, cell death and regeneration in the adult gymnotiform brain.

Gymnotiform fish, like all teleosts examined thus far, are distinguished by their enormous potential for the production of new neurons in the adult brain. In Apteronotus leptorhynchus, on average 10(5) cells, corresponding to approximately 0.2 % of the total population of cells in the adult brain, are in S-phase within any period of 2 h. At least a portion of these newly generated cells survive for the rest of the fish's life. This long-term survival, together with the persistent generation of new cells, leads to a continuous growth of the brain during adulthood. Zones of high proliferative activity are typically located at or near the surface of the ventricular, paraventricular and cisternal systems. In the central posterior/ prepacemaker nucleus, for example, new cells are generated, at very high rates, in areas near the wall of the third ventricle. At least some of these cells differentiate into neurons, express immunoreactivity against the neuropeptide somatostatin and migrate into more lateral areas of this complex. Approximately 75 % of all new brain cells are generated in the cerebellum. In the corpus cerebelli and the valvula cerebelli, they are produced in the molecular layers, whereas in the eminentia granularis the newborn cells stem from proliferation zones in the pars medialis. Within the first few days of their life, these cells migrate towards specific target areas, namely the associated granule cell layers. At least some of them develop into granule neurons. The high proliferative activity is counterbalanced by apoptosis, a mechanism that resembles the processes known from embryonic development of the vertebrate brain. Apoptosis also appears to be used as an efficient mechanism for the removal of cells damaged through injury in the brain of adult Apteronotus leptorhynchus. Since apoptosis is not accompanied by the side effects known from necrosis, this 'clean' type of cell death may, together with the enormous proliferative activity in the brain, explain, at least partially, the tremendous capability of teleost fish to replace damaged neurons with newly generated ones. One factor that appears to play a major role in the generation of new cells and in their further development is the neuropeptide somatostatin. In the caudal cerebellum of the gymnotiform brain, somatostatin-binding sites are expressed, at extremely high densities, at sites corresponding to the areas of origin, migration and differentiation of the newborn cells. This pattern of expression resembles the expression pattern in the rat cerebellum, where somatostatin immunoreactivity and somatostatin-binding sites are transiently expressed at the time when the granule cells of the cerebellum are generated. Moreover, after mechanical lesions of the corpus cerebelli, the expression of somatostatin-like immunoreactivity is tremendously increased in several cell types (presumably astrocytes, microglia and granule cell neurons) near the path of the lesion; the time course of this expression coincides with the temporal pattern underlying the recruitment of new cells incorporated at the site of the lesion.

Plasticity of feedback inputs in the apteronotid electrosensory system.

Weakly electric fish generate an electric field surrounding their body by means of an electric organ typically located within the trunk and tail. Electroreceptors scattered over the surface of the body encode the amplitude and timing of the electric organ discharge (EOD), and central components of the electrosensory system analyze the information provided by the electroreceptor afferents. The electrosensory system is used for electrolocation, for the detection and analysis of objects near the fish which distort the EOD and for electrocommunication. Since the electric organ is typically located in the tail, any movement of this structure relative to the rest of the body alters the EOD field, resulting in large changes in receptor afferent activity. The amplitude of these reafferent stimuli can exceed the amplitudes of near-threshold electrolocation signals by several orders of magnitude. This review summarizes recent studies of the South American weakly electric fish Apteronotus leptorhynchus aimed at determining how the animals differentiate self-generated or reafferent electrosensory stimuli from those that are more behaviorally relevant. Cells within the earliest stages of central electrosensory processing utilize an adaptive filtering technique which allows the system preferentially to attenuate reafferent as well as other predictable patterns of sensory input without degrading responses to more novel stimuli. Synaptic plasticity within the system underlies the adaptive component of the filter and enables the system to learn to reject new stimulus patterns if these become predictable. A Ca2+-mediated form of postsynaptic depression contributes to this synaptic plasticity. The filter mechanism seen in A. leptorhynchus is surprisingly similar to adaptive filters described previously in mormyrid weakly electric fish and in elasmobranchs, suggesting that this mechanism may be a common feature of sensory processing systems.

Molecular biology of the apteronotus NMDA receptor NR1 subunit.

The complete sequences and expression patterns of the NR1 (aptNR1) subunit of the N-methyl-d-aspartate (NMDA) receptor and its alternative splice isoforms have been determined for the weakly electric fish Apteronotus leptorhynchus. The deduced amino acid sequence of aptNR1 is approximately 88 % identical to the NR1 sequences of other vertebrate. Two of the three alternative splice cassettes previously described for mammalian NR1s, N1 and C1, are present in aptNR1, but the third cassette, C2, is not found. In addition, two teleost-specific splice cassettes occur on the N-terminal side of the C1 sequence. The cellular patterns of aptNR1 expression, including the patterns of N1 and C1 splicing, have been mapped using the in situ hybridization technique. High levels of aptNR1 mRNA were detected throughout the central nervous system including most neurons of the electrosensory system, with the highest levels in electrosensory lateral line lobe pyramidal cells. Expression of the N1 splice isoform was higher in more caudal regions of the brain, and expression of the C1 splice isoform was higher in more rostral regions. The N1 splice isoform was present in almost all NR1-positive cells, in contrast to the C1 splice isoform which was restricted to a subset of NR1-positive cells. These results demonstrate that the NR1 subunit of the NMDA receptor is evolutionarily conserved across species and that regulation of alternative RNA splicing modulates the properties of NR1 in different neurons of the central nervous system of A. leptorhynchus.

Testosterone increases the number of substance P-like immunoreactive neurons in a specific sub-division of the lateral hypothalamus of the weakly electric, brown ghost knifefish, Apteronotus leptorhynchus.

During spawning, male and female brown ghost knife fish modulate their electric organ discharge to produce discrete courtship signals known as chirps. However, non-spawning fish show clear sex differences in chirp responsiveness to electrosensory stimuli; males consistently chirp, whereas females do not. This behavioral dimorphism is paralleled by sex differences in substance P-like immunoreactivity (SPl-ir) in the prepacemaker nucleus (PPn) which regulates chirping. The PPn is densely innervated by SPl-ir fibers in males, but not in females. However, we have shown that both female chirping behavior and the expression of SPl-ir in the PPn are enhanced following adult testosterone treatment [J.G. Dulka, L. Maler, W. Ellis, Androgen-induced changes in electrocommunicatory behavior are correlated with changes in substance P-like immunoreactivity in the brain of the electric fish, Apteronotus leptorhynchus, J. Neurosci. 15 (1995) 1879-1890]. Thus, testosterone-induced changes in SPl-ir input to the PPn may modulate female chirping during spawning. Recent evidence suggests that SPl-ir projections to the PPn may originate from SPl-ir neurons in the lateral hypothalamus (Hl). If so, one might expect to see a greater number of SPl-ir neurons in the Hl of testosterone-implanted females compared to Blank-implanted controls. In making this comparison, we found that both groups of females had similar numbers of SPl-ir neurons in the anterior Hl, but that testosterone-implanted females had significantly (p<0.01) more SPl-ir neurons in a distinct part of the posterior Hl. This sub-population of testosterone-sensitive, SPl-ir neurons may contribute to the regulation of chirping, since an increase in their number is positively correlated with the appearance of SPl-ir fibers in the PPn [J.G. Dulka, L. Maler, W. Ellis, Androgen-induced changes in electrocommunicatory behavior are correlated with changes in substance P-like immunoreactivity in the brain of the electric fish, Apteronotus leptorhynchus, J. Neurosci. 15 (1995) 1879-1890].

Inhibition evoked from primary afferents in the electrosensory lateral line lobe of the weakly electric fish (Apteronotus leptorhynchus).

Inhibition evoked from primary afferents in the electrosensory lateral line lobe of the weakly electric fish (Apteronotus leptorhynchus). J. Neurophysiol. 80: 3173-3196, 1998. The responses of two types of projection neurons of the electrosensory lateral line lobe, basilar (BP) and nonbasilar (NBP) pyramidal cells, to stimulation of primary electrosensory afferents were determined in the weakly electric fish, Apteronotus leptorhynchus. Using dyes to identify cell type, the response of NBP cells to stimulation of primary afferents was inhibitory, whereas the response of BP cells was excitation followed by inhibition. gamma-Aminobutyric acid (GABA) applications produced biphasic (depolarization then hyperpolarization) responses in most cells. GABAA antagonists blocked the depolarizing effect of GABA and reduced the hyperpolarizing effect. The GABAB antagonists weakly antagonized the hyperpolarizing effect. The early depolarization had a larger increase in cell conductance than the late hyperpolarization. The conductance changes were voltage dependent, increasing with depolarization. In both cell types, baclofen produced a slow small hyperpolarization and reduced the inhibitory postsynaptic potentials (IPSPs) evoked by primary afferent stimulation. Tetanic stimulation of primary afferents at physiological rates (100-200 Hz) produced strongly summating compound IPSPs (approximately 500-ms duration) in NBP cells, which were usually sensitive to GABAA but not GABAB antagonists; in some cells there remained a slow IPSP that was unaffected by GABAB antagonists. BP cells responded with excitatory or mixed excitatory + inhibitory responses. The inhibitory response had both a fast (approximately 30 ms, GABAA) and long-lasting slow phase (approximately 800 ms, mostly blocked by GABAA antagonists). In some cells there was a GABAA antagonist-insensitive slow IPSP (approximately 500 ms) that was sensitive to GABAB antagonists. Application of glutamate ionotropic receptor antagonists blocked the inhibitory response of NBP cells to primary afferent stimulation and the excitatory response of BP cells but enhanced the BP cell slow IPSP; this remaining slow IPSP was reduced by GABAB antagonists. Unit recordings in the granule cell layer and computer simulations of pyramidal cell inhibition suggested that the duration of the slow GABAA inhibition reflects the prolonged firing of GABAergic granule cell interneurons to primary afferent input. Correlation of the results with known GABAergic circuitry in the electrosensory lobe suggests that the GABAergic type 2 granule cell input to both pyramidal cell types is via GABAA receptors. The properties of the GC2 GABAA input are well suited to their putative role in gain control, regulation of phasicness, and coincidence detection. The slow GABAB IPSP evoked in BP cells is likely due to ovoid cell input to their basal dendrites.

Differential roles of Ca2+/calmodulin-dependent kinases in posttetanic potentiation at input selective glutamatergic pathways.

The electrosensory lateral line lobe (ELL) of the electric fish Apteronotus leptorhynchus is a layered medullary region receiving electroreceptor input that terminates on basal dendrites of interneurons and projection (pyramidal) cells. The molecular layer of the ELL contains two distinct glutamatergic feedback pathways that terminate on the proximal (ventral molecular layer, VML) and distal (dorsal molecular layer) apical dendrites of pyramidal cells. Western blot analysis with an antibody directed against mammalian Ca2+/calmodulin-dependent kinase 2, alpha subunit (CaMK2alpha) recognized a protein of identical size in the brain of A. leptorhynchus. Immunohistochemistry demonstrated that CaMK2 alpha expression in the ELL was restricted to fibers and terminals in the VML. Posttetanic potentiation (PTP) could be readily elicited in pyramidal cells by stimulation of either VML or DML in brain slices of the ELL. PTP in the VML was blocked by extracellular application of a CaMK2 antagonist (KN62) while intracellular application of KN62 or a CaMK2 inhibitory peptide had no effect, consistent with the presynaptic localization of CaMK2 alpha in VML. PTP in the dorsal molecular layer was not affected by extracellular application of KN62. Anti-Hebbian plasticity has also been demonstrated in the VML, but was not affected by KN62. These results demonstrate that, while PTP can occur independent of CaMK2, it is, in some synapses, dependent on this kinase.

Diversity of sexual dimorphism in electrocommunication signals and its androgen regulation in a genus of electric fish, Apteronotus.

Gymnotiform electric fish emit an electric organ discharge that, in several species, is sexually dimorphic and functions in gender recognition. In addition, some species produce frequency modulations of the electric organ discharge, known as chirps, that are displayed during aggression and courtship. We report that two congeneric species (Apteronotus leptorhynchus and A. albifrons) differ in the expression of sexual dimorphism in these signals. In A. leptorhynchus, males chirp more than females, but in A. albifrons chirping is monomorphic. The gonadosomatic index and plasma levels of 11-ketotestosterone were equivalent in both species, suggesting that they were in similar reproductive condition. Corresponding to this difference in dimorphism, A. leptorhynchus increases chirping in response to androgens, but chirping in A. albifrons is insensitive to implants of testosterone, dihydrotestosterone or 11-ketotestosterone. Species also differ in the sexual dimorphism and androgen sensitivity of electric organ discharge frequency. In A. leptorhynchus, male discharge at higher frequencies than females, and androgens increase electric organ discharge frequency. In A. albifrons, males discharge at lower frequencies than females, and androgens decrease electronic organ discharge frequency. Thus, in both chirping and electric organ discharge frequency, evolutionary changes in the presence or direction of sexual dimorphism have been accompanied and perhaps caused by changes in the androgen regulation of the electric organ discharge.

Testing the order of discrete Markov chains using surrogate data

Tests of the null-hypothesis that a symbolic sequence is of nth Markov order are described. These tests are based on resampling methods similar to the method of surrogate data used to test continuous time series for nonlinear structure. The size and power of these symbolic surrogate tests are estimated using binary and heptary test-sequences of known order. Conditioning on a sufficient statistic under the null-hypothesis yields an estimated true size which closely approximates the nominal size and increases the power of the test with respect to the unconditional test. We apply one method to a symbolic sequence derived from the Rössler system and to a binary and heptary representation of an interspike interval sequence of an electroreceptor of the weakly electric eel Apteronotus Leptorhynchus.

Sequence diversity of voltage-gated potassium channels in an electric fish

Cloning of voltage-gated K + channels has indicated that these channels constitute a diverse family of genes that have been subclassified into four closely related gene families Kv1–Kv4 ( Shaker, Shab, Shaw, Shal). A PCR approach has been used to assess the diversity of K + channels in the weakly electric fish Apteronotus leptorhynchus, which is a well studied model of sensory processing. Degenerate primers specific for the highly conserved pore and S6 transmembrane domains of the K + channel families were used to amplify an intronless 124 bp fragment from fish genomic DNA. DNA sequence analysis of a large number of these fragments has identified 19 putative K + channels, each of which can be classified into one of the four major families. Ten fall into the Kv1 class, two in the Kv2 class, five in the Kv3 class and two in the Kv4 class. The results indicate that the duplications that gave rise to multiple genes within each of the K + channel families predate the divergence of the Actinopterygii and Sarcopterygii lineages (∼400 million years ago) during early vertebrate evolution.

N-methyl-D-aspartate receptor 1 mRNA distribution in the central nervous system of the weakly electric fish Apteronotus leptorhynchus.

We have isolated a partial cDNA for the N-methyl-D-aspartate (NMDA) receptor 1 (NMDAR1) subunit from an Apteronotus leptorhynchus brain cDNA library. The A. leptorhynchus cDNA fragment, which corresponds to nucleotides 135-903 within the 5' region of the rat NR1 mRNA, encodes 252 amino acids that are >80% identical to the homologous segments of the rat, human, and duck NR1 proteins. RNAse protection assays revealed that the A. leptorhynchus NR1 mRNA was highly enriched in the forebrain and hypothalamus, with lesser amounts in the brainstem, and very low levels in the cerebellum. In situ hybridization also demonstrated that neurons in the pallial forebrain were highly enriched in NR1 transcripts. High levels of NR1 mRNA were found in pyramidal cells within the optic tectum and octavolateral regions. Pyramidal cells of the electrosensory lateral line lobe had the highest levels of expression, and the NR1 mRNA was found to be selectively enriched in their apical dendrites.