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Unlike mammals, species such as fish and amphibians can regenerate damaged spinal cords, offering insights into potential therapeutic targets. This study investigates the structural features of the molly fish spinal cord through light and electron microscopy. The most notable characteristic was the presence of Mauthner cells (M‐cells), which exhibited large cell bodies and processes, as well as synaptic connections with astrocytes. These astrocytic connections contained synaptic vesicles, suggesting electrical transmission at the M‐cell endings. Astrocytes, which were labeled with glial fibrillary acidic protein (GFAP), contained cytoplasmic glycogen granules, potentially serving as an emergency fuel source. Two types of oligodendrocytes were identified: a small, dark cell and a larger, lighter cell, both of which reacted strongly with oligodendrocyte transcription factor 2 (Olig2). The dark oligodendrocyte resembled human oligodendrocyte precursors, while the light oligodendrocyte was similar to mature human oligodendrocytes. Additionally, proliferative neurons in the substantia grisea centralis expressed myostatin, Nrf2, and Sox9. Collectively, these findings suggest that the molly fish spinal cord has advanced structural features conducive to spinal cord regeneration and could serve as an excellent model for studying central nervous system regeneration. Further studies on the functional aspects of the molly fish spinal cord are recommended. Research Highlights: Mauthner cells (M‐cell), with their typical large cell body and processes, were the most characteristic feature in Molly fish spinal cord, where it presented synaptic connections with astrocytes and their ends contained synaptic vesicles indicating an electrical transmission in the M‐cells endings.Two types of oligodendrocytes could be recognized; both reacted intensely with Oligodendrocyte transcription factor 2 (Olig2).The proliferative neurons of the substantia grisea centralis expressed myostatin, Nrf2, and Sox9.The findings of this study suggest that molly fish possess highly developed structural features conducive to spinal cord regeneration. Consequently, they could be deemed an exemplary model for investigating central nervous system regeneration. [ABSTRACT FROM AUTHOR]

In animal species ranging from invertebrate to mammals, visually guided escape behaviours have been studied using looming stimuli, the two‐dimensional expanding projection on a screen of an object approaching on a collision course at constant speed. The peak firing rate or membrane potential of neurons responding to looming stimuli often tracks a fixed threshold angular size of the approaching stimulus that contributes to the triggering of escape behaviours. To study whether this result holds more generally, we designed stimuli that simulate acceleration or deceleration over the course of object approach on a collision course. Under these conditions, we found that the angular threshold conveyed by collision detecting neurons in grasshoppers was sensitive to acceleration whereas the triggering of escape behaviours was less so. In contrast, neurons in goldfish identified through the characteristic features of the escape behaviours they trigger, showed little sensitivity to acceleration. This closely mirrored a broader lack of sensitivity to acceleration of the goldfish escape behaviour. Thus, although the sensory coding of simulated colliding stimuli with non‐zero acceleration probably differs in grasshoppers and goldfish, the triggering of escape behaviours converges towards similar characteristics. Approaching stimuli with non‐zero acceleration may help refine our understanding of neural computations underlying escape behaviours in a broad range of animal species. Key points: A companion manuscript showed that two mathematical models of collision‐detecting neurons in grasshoppers and goldfish make distinct predictions for the timing of their responses to simulated objects approaching on a collision course with non‐zero acceleration.Testing these experimental predictions showed that grasshopper neurons are sensitive to acceleration while goldfish neurons are not, in agreement with the distinct models proposed previously in these species using constant velocity approaches.Grasshopper and goldfish escape behaviours occurred after the stimulus reached a fixed angular size insensitive to acceleration, suggesting further downstream processing in grasshopper motor circuits to match what was observed in goldfish.Thus, in spite of different sensory processing in the two species, escape behaviours converge towards similar solutions.The use of object acceleration during approach on a collision course may help better understand the neural computations implemented for collision avoidance in a broad range of species. [ABSTRACT FROM AUTHOR]

The processing of visual information for collision avoidance has been investigated at the biophysical level in several model systems. In grasshoppers, the (so-called) η model captures reasonably well the visual processing performed by an identified neuron called the lobular giant movement detector as it tracks approaching objects. Similar phenomenological models have been used to describe either the firing rate or the membrane potential of neurons responsible for visually guided collision avoidance in other animals. Specifically, in goldfish, the κ model has been proposed to describe the Mauthner cell, an identified neuron involved in startle escape responses. In the vinegar fly, a third model was developed for the giant fiber neuron, which triggers last resort escapes immediately before an impending collision. One key property of these models is their prediction that peak neuronal responses occur at a fixed delay after the simulated approaching object reaches a threshold angular size on the retina. This prediction is valid for simulated objects approaching at a constant speed. We tested whether it remains valid when approaching objects accelerate. After characterizing and comparing the models' responses to accelerating and constant speed stimuli, we find that the prediction holds true for the κ and the giant fiber model, but not for the η model. These results suggest that acceleration in the approach trajectory of an object may help distinguish and further constrain the neuronal computations required for collision avoidance in grasshoppers, fish and vinegar flies. [ABSTRACT FROM AUTHOR]

Wolfram Syndrome (WS) is a fatal human inherited disease with symptoms of diabetes, vision decreasing, and neurodegeneration caused by mutations in the endoplasmic reticulum (ER)-resident protein WFS1. WFS1 has been reported to play an important role in glucose metabolism. However, the role of WFS1 in axonal regeneration in the central nervous system has so far remained elusive. Herein, we established a model of the wfs1b globally deficient zebrafish line. wfs1b deficiency severely impeded the Mauthner-cell (M-cell) axon regeneration, which was partly dependent on the ER stress response. The administration of ER stress inhibitor 4-Phenylbutyric acid (4-PBA) promoted M-cell axon regeneration in wfs1b−/− zebrafish larvae, while the ER stress activator Tunicamycin (TM) inhibited M-cell axon regeneration in wfs1b+/+ zebrafish larvae. Moreover, complementation of wfs1b at the single-cell level stimulated M-cell axon regeneration in the wfs1b−/− zebrafish larvae. Altogether, our results revealed that wfs1b promotes M-cell axon regeneration through the ER stress signal pathway and provide new evidence for a therapeutic target for WS and axon degeneration. [ABSTRACT FROM AUTHOR]

Social status-dependent modulation of neural circuits has been investigated extensively in vertebrate and invertebrate systems. However, the effects of social status on neuromodulatory systems that drive motor activity are poorly understood. Zebrafish form a stable social relationship that consists of socially dominant and subordinate animals. The locomotor behavior patterns differ according to their social ranks. The sensitivity of the Mauthner startle escape response in subordinates increases compared to dominants while dominants increase their swimming frequency compared to subordinates. Here, we investigated the role of the endocannabinoid system (ECS) in mediating these differences in motor activities. We show that brain gene expression of key ECS protein pathways are socially regulated. Diacylglycerol lipase (DAGL) expression significantly increased in dominants and significantly decreased in subordinates relative to controls. Moreover, brain gene expression of the cannabinoid 1 receptor (CB1R) was significantly increased in subordinates relative to controls. Secondly, increasing ECS activity with JZL184 reversed swimming activity patterns in dominant and subordinate animals. JZL184 did not affect the sensitivity of the startle escape response in dominants while it was significantly reduced in subordinates. Thirdly, blockage of CB1R function with AM-251 had no effect on dominants startle escape response sensitivity, but startle sensitivity was significantly reduced in subordinates. Additionally, AM-251 did not affect swimming activities in either social phenotypes. Fourthly, we demonstrate that the effects of ECS modulation of the startle escape circuit is mediated via the dopaminergic system specifically via the dopamine D1 receptor. Finally, our empirical results complemented with neurocomputational modeling suggest that social status influences the ECS to regulate the balance in synaptic strength between excitatory and inhibitory inputs to control the excitability of motor behaviors. Collectively, this study provides new insights of how social factors impact nervous system function to reconfigure the synergistic interactions of neuromodulatory pathways to optimize motor output. [ABSTRACT FROM AUTHOR]

Dual oxidase (duox)-derived reactive oxygen species (ROS) have been correlated with neuronal polarity, cerebellar development, and neuroplasticity. However, there have not been many comprehensive studies of the effect of individual duox isoforms on central-axon regeneration in vivo. Here, we explored this question in zebrafish, an excellent model organism for central-axon regeneration studies. In our research, mutation of the duox gene with CRISPR/Cas9 significantly retarded the single-axon regeneration of the zebrafish Mauthner cell in vivo. Using deep transcriptome sequencing, we found that the expression levels of related functional enzymes in mitochondria were down-regulated in duox mutant fish. In vivo imaging showed that duox mutants had significantly disrupted mitochondrial transport and redox state in single Mauthner-cell axon. Our research data provide insights into how duox is involved in central-axon regeneration by changing mitochondrial transport. [ABSTRACT FROM AUTHOR]

During many behaviors in vertebrates, the CNS generates asymmetric activities between the left and right sides to produce asymmetric body movements. For asymmetrical activations of the CNS, reciprocal inhibition between the left and right sides is believed to play a key role. However, the complexity of the CNS makes it difficult to identify the reciprocal inhibition circuits at the level of individual cells and the contribution of each neuron to the asymmetric activity. Using larval zebrafish, we examined this issue by investigating reciprocal inhibition circuits between a pair of Mauthner (M) cells, giant reticulospinal neurons that trigger fast escapes. Previous studies have shown that a class of excitatory neurons, called cranial relay neurons, is involved in the reciprocal inhibition pathway between the M cells. Using transgenic fish, in which two of the cranial relay neurons (Ta1 and Ta2) expressed GFP, we showed that Ta1 and Ta2 constitute major parts of the pathway. In larvae in which Ta1/Ta2 were laser-ablated, the amplitude of the reciprocal IPSPs dropped to less than one-third. Calcium imaging and electrophysiological recording showed that the occurrence probability of bilateral M-cell activation upon sound/vibration stimuli was greatly increased in the Ta1/Ta2-ablated larvae. Behavioral experiments revealed that the Ta1/Ta2 ablation resulted in shallower body bends during sound/vibration-evoked escapes, which is consistent with the observation that increased occurrence of bilateral M-cell activation impaired escape performance. Our study revealed major components of the reciprocal inhibition circuits in the M cell system and the behavioral importance of the circuits. [ABSTRACT FROM AUTHOR]

Key points: The present study examines dendritic integrative processes that occur in many central neurons but have been challenging to study in vivo in the vertebrate brain. The Mauthner cell of goldfish receives auditory and visual information via two separate dendrites, providing a privileged scenario for in vivo examination of dendritic integration. The results show differential attenuation properties in the Mauthner cell dendrites arising at least partly from differences in cable properties and the nonlinear behaviour of the respective dendritic membranes. In addition to distinct modality‐dependent membrane specialization in neighbouring dendrites of the Mauthner cell, we report cross‐modal dendritic interactions via backpropagating postsynaptic potentials. Broadly, the results of the present study provide an exceptional example for the processing power of single neurons. Abstract: Animals process multimodal information for adaptive behavioural decisions. In fish, evasion of a diving bird that breaks the water surface depends on integrating visual and auditory stimuli with very different characteristics. How do neurons process such differential sensory inputs at the dendritic level? For that, we studied the Mauthner cells (M‐cells) in the goldfish startle circuit, which receive visual and auditory inputs via two separate dendrites, both accessible for in vivo recordings. We investigated whether electrophysiological membrane properties and dendrite morphology, studied in vivo, play a role in selective sensory processing in the M‐cell. The results obtained show that anatomical and electrophysiological differences between the dendrites combine to produce stronger attenuation of visually evoked postsynaptic potentials (PSPs) than to auditory evoked PSPs. Interestingly, our recordings showed also cross‐modal dendritic interaction because auditory evoked PSPs invade the ventral dendrite (VD), as well as the opposite where visual PSPs invade the lateral dendrite (LD). However, these interactions were asymmetrical, with auditory PSPs being more prominent in the VD than visual PSPs in the LD. Modelling experiments imply that this asymmetry is caused by active conductances expressed in the proximal segments of the VD. The results obtained in the present study suggest modality‐dependent membrane specialization in M‐cell dendrites suited for processing stimuli of different time domains and, more broadly, provide a compelling example of information processing in single neurons. [ABSTRACT FROM AUTHOR]

Electrical synapses are an omnipresent feature of nervous systems, from the simple nerve nets of cnidarians to complex brains of mammals. Formed by gap junction channels between neurons, electrical synapses allow direct transmission of voltage signals between coupled cells. The relative simplicity of this arrangement belies the sophistication of these synapses. Coupling via electrical synapses can be regulated by a variety of mechanisms on times scales ranging from milliseconds to days, and active properties of the coupled neurons can impart emergent properties such as signal amplification, phase shifts and frequency-selective transmission. This article reviews the biophysical characteristics of electrical synapses and some of the core mechanisms that control their plasticity in the vertebrate central nervous system. [ABSTRACT FROM AUTHOR]

Opioid receptors were shown to modulate a variety of cellular processes in the vertebrate central nervous system, including synaptic transmission. While the effects of opioid receptors on chemically mediated transmission have been extensively investigated, little is known of their actions on gap junction-mediated electrical synapses. Here we report that pharmacological activation of mu-opioid receptors led to a long-term enhancement of electrical (and glutamatergic) transmission at identifiable mixed synapses on the goldfish Mauthner cells. The effect also required activation of both dopamine D1/5 receptors and postsynaptic cAMP-dependent protein kinase A, suggesting that opioid-evoked actions are mediated indirectly via the release of dopamine from varicosities known to be located in the vicinity of the synaptic contacts. Moreover, inhibitory inputs situated in the immediate vicinity of these excitatory synapses on the lateral dendrite of the Mauthner cell were not affected by activation of mu-opioid receptors, indicating that their actions are restricted to electrical and glutamatergic transmissions co-existing at mixed contacts. Thus, as their chemical counterparts, electrical synapses can be a target for the modulatory actions of the opioid system. Because gap junctions at these mixed synapses are formed by fish homologs of the neuronal connexin 36, which is widespread in mammalian brain, it is likely that this regulatory property applies to electrical synapses elsewhere as well. [ABSTRACT FROM AUTHOR]

Prepulse inhibition (PPI) is understood as a sensorimotor gating process that attenuates sensory flow to the startle pathway during early stages (20--1000 ms) of information processing. Here, we applied in vivo electrophysiology and pharmacology to determine if PPI is mediated by glycine receptors (GlyRs) and/or GABAA receptors (GABAARs) in the goldfish auditory startle circuit. Specifically, we used selective antagonists to dissect the contributions of target receptors on sound-evoked postsynaptic potentials (PSPs) recorded in the neurons that initiate startle, the Mauthner-cells (M-cell). We found that strychnine, a GlyR antagonist, disrupted a fast-activated (5 ms) and rapidly (<50 ms) decaying (feed-forward) inhibitory process that contributes to PPI at 20 ms prepulse/pulse inter-stimulus intervals (ISI). Additionally we observed increases of the evoked postsynaptic potential (PSP) peak amplitude (+87.43 ± 21.53%, N = 9) and duration (+204 ± 48.91%, N = 9). In contrast, treatment with bicuculline, a GABAAR antagonist, caused a general reduction in PPI across all tested interstimulus intervals (ISIs) (20-500 ms). Bicuculline also increased PSP peak amplitude (+133.8 ± 10.3%, N = 5) and PSP duration (+284.95 ± 65.64%, N = 5). Treatment with either antagonist also tonically increased post-synaptic excitability in the M-cells, reflected by an increase in the magnitude of antidromically-evoked action potentials (APs) by 15.07 ± 3.21%, N = 7 and 16.23 ± 7.08%, N = 5 for strychnine and bicuculline, respectively. These results suggest that GABAARs and GlyRs are functionally segregated to short- and longer-lasting sound-evoked (phasic) inhibitory processes that contribute to PPI, with the mediation of tonic inhibition by both receptor systems being critical for gain control within the M-cell startle circuit. [ABSTRACT FROM AUTHOR]

In contrast to the knowledge of chemical synapses, little is known regarding the properties of gap junction-mediated electrical synapses in developing zebrafish, which provide a valuable model to study neural function at the systems level. Identifiable "mixed" (electrical and chemical) auditory synaptic contacts known as "club endings" on Mauthner cells (2 large reticulospinal neurons involved in tail-flip escape responses) allow exploration of electrical transmission in fish. Here, we show that paralleling the development of auditory responses, electrical synapses at these contacts become anatomically identifiable at day 3 postfertilization, reaching a number of~6 between days 4 and 9. Furthermore, each terminal contains ~18 gap junctions, representing between 2,000 and 3,000 connexon channels formed by the teleost homologs of mammalian connexin 36. Electrophysiological recordings revealed that gap junctions at each of these contacts are functional and that synaptic transmission has properties that are comparable with those of adult fish. Thus a surprisingly small number of mixed synapses are responsible for the acquisition of auditory responses by the Mauthner cells, and these are likely sufficient to support escape behaviors at early developmental stages. [ABSTRACT FROM AUTHOR]

Rapid escape swims in fish are initiated by the Mauthner cells, giant reticulospinal neurons with unique specializations for swift responses. The Mauthner cells directly activate motoneurons and facilitate predator detection by integrating acoustic, mechanosensory, and visual stimuli. In addition, larval fish show well-coordinated escape responses when exposed to electric field pulses (EFPs). Sensitization of the Mauthner cell by genetic overexpression of the voltage-gated sodium channel SCN5 increased EFP responsiveness, whereas Mauthner ablation with an engineered variant of nitroreductase with increased activity (epNTR) eliminated the response. The reaction time to EFPs is extremely short, with many responses initiated within 2 ms of the EFP. Large neurons, such as Mauthner cells, show heightened sensitivity to extracellular voltage gradients. We therefore tested whether the rapid response to EFPs was due to direct activation of the Mauthner cells, bypassing delays imposed by stimulus detection and transmission by sensory cells. Consistent with this, calcium imaging indicated that EFPs robustly activated the Mauthner cell but only rarely fired other reticulospinal neurons. Further supporting this idea, pharmacological blockade of synaptic transmission in zebrafish did not affect Mauthner cell activity in response to EFPs. Moreover, Mauthner cells transgenically expressing a tetrodotoxin (TTX)-resistant voltage-gated sodium channel retained responses to EFPs despite TTX suppression of action potentials in the rest of the brain. We propose that EFPs directly activate Mauthner cells because of their large size, thereby driving ultrarapid escape responses in fish. [ABSTRACT FROM AUTHOR]

Each neuron possesses a unique firing property, which is largely attributed to heterogeneity in the composition of voltage-gated ion channel complexes. Zebrafish Mauthner (M) cells, which are bilaterally paired giant reticulospinal neurons (RSNs) in the hindbrain and induce rapid escape behavior, generate only a single spike at the onset of depolarization. This single spiking is in contrast with the repetitive firing of the M cell's morphologically homologous RSNs, MiD2cm and MiD3cm, which are also involved in escapes. However, how the unique firing property of M cells is established and the underlying molecular mechanisms remain unclear. In the present study, we first demonstrated that the single-spiking property of M cells was acquired at 4 days postfertilization (dpf), accompanied by an increase in dendrotoxin I (DTX)-sensitive low-threshold K+ currents, prior to which the M cell repetitively fires as its homologs. Second, in situ hybridization showed that among DTX-sensitive Kv1 channel α-subunits, zKv1.1a was unexpectedly expressed even in the homologs and the bursting M cells at 2 dpf. In contrast, zKvβ2b, an auxiliary β-subunit of Kv1 channels, was expressed only in the single-spiking M cells. Third, zKv1.1a expressed in Xenopus oocytes functioned as a low-threshold K+ channel, and its currents were enhanced by coexpression of zKvβ2b subunits. Finally, knockdown of zKvβ2b expression in zebrafish larvae resulted in repetitive firing of M cells at 4 dpf. Taken together, these results suggest that associative expression of Kvβ2 subunits with Kv1.1 channels is crucial for developmental acquisition of the unique firing properties of the M cells among homologous neurons. [ABSTRACT FROM AUTHOR]

Segmental organization along the neuraxis is a prominent feature of the CNS in vertebrates. In a wide range of fishes, hindbrain segments contain orderly arranged reticulospinal neurons (RSNs). Individual RSNs in goldfish and zebrafish hindbrain are morphologically identified. RSNs sharing similar morphological features are called segmental homologs and repeated in adjacent segments. However, little is known about functional relationships among segmental homologs. Here we investigated the electrophysiological connectivity between the Mauthner cell (M-cell), a pair of giant RSNs in segment 4 (r4) that are known to trigger fast escape behavior, and different series of homologous RSNs in r4 -r6. Paired intracellular recordings in adult goldfish revealed unidirectional connections from the M-cell to RSNs. The connectivity was similar in morphological homologs. A single M-cell spike produced IPSPs in dorsally located RSNs (MiD cells) on the ipsilateral side and excitatory postsynaptic depolarization on the contralateral side, except for MiD2cm cells. The inhibitory or excitatory potentials effectively suppressed or enhanced target RSNs spiking, respectively. In contrast to the lateralized effects on MiD cells, single M-cell spiking elicited equally strong depolarizations on bilateral RSNs located ventrally (MiV cells), and the depolarization was high enough for MiV cells to burst. Therefore, the morphological homology of repeated RSNs in r4 -r6 and their functional M-cell connectivity were closely correlated, suggesting that each functional connection works as a functional motif during the M-cellinitiated escape. [ABSTRACT FROM AUTHOR]

Locomotor feedback signals from the spinal cord to descending brain stem neurons were examined in the lamprey using the uniquely identifiable reticulospinal neurons, the Müller and Mauthner cells. The same identified reticulospinal neurons were recorded in several preparations, under reduced conditions, to address whether an identified reticulospinal neuron shows similar locomotor-related oscillation timing from animal to animal and whether these timing signals can differ significantly from other identified reticulospinal neurons. Intracellular recordings of membrane potential in identified neurons were made in an isolated brain stem-spinal cord preparation with a high-divalent cation solution on the brain stem to suppress indirect neural pathways and with D-glutamate perfusion to the spinal cord to induce fictive swimming. Under these conditions, the identified reticulospinal neurons show significant clustering of the timings of the peaks and troughs of their locomotor-related oscillations. Whereas most identified neurons oscillated in phase with locomotor bursting in ipsilateral ventral roots of the rostral spinal cord, the B1 Müller cell, which has an ipsilateral descending axon, and the Mauthner cell, which has a contralateral descending axon, both had oscillation peaks that were out of phase with the ipsilateral ventral roots. The differences in oscillation timing appear to be due to differences in synaptic input sources as shown by cross-correlations of fast synaptic activity in pairs of Müller cells. Since the main source of the locomotor input under these experimental conditions is ascending neurons in the spinal cord, these experiments suggest that individual reticulospinal neurons can receive locomotor signals from different subsets of these ascending neurons. This result may indicate that the locomotor feedback signals from the spinal locomotor networks are matched in some way to the motor output functions of the individual reticulospinal neurons, which include command signals for turning and for compensatory movements. [ABSTRACT FROM AUTHOR]

Social life affects brain function at all levels, including gene expression, neurochemical balance, and neural circuits. We have previously shown that in the cichlid fish Astatotilapia burtoni brightly colored, socially dominant (DOM) males face a trade-off between reproductive opportunities and increased predation risk. Compared with camouflaged subordinate (SUB) males, DOMs exposed to a loud sound pip display higher startle responsiveness and increased excitability of the Mauthner cell (M-cell) circuit that governs this behavior. Using behavioral tests, intracellular recordings, and single-cell molecular analysis, we show here that serotonin (5-HT) modulates this socially regulated plasticity via the 5-HT receptor subtype 2 (5-HTR2). Specifically, SUBs display increased sensitivity to pharmacological manipulation of 5-HTR2 compared with DOMs in both startle-escape behavior and electrophysiological properties of the M-cell. Immunohistochemistry showed serotonergic varicosities around the M-cells, further suggesting that 5-HT impinges directly onto the startle-escape circuitry. To determine whether the effects of 5-HTR2 are pre- or postsynaptic, and whether other 5-HTR subtypes are involved, we harvested the mRNA from single M-cells via cytoplasmic aspiration and found that 5-HTR subtypes 5A and 6 are expressed in the M-cell. 5-HTR2, however, was absent, suggesting that it affects M-cell excitability through a presynaptic mechanism. These results are consistent with a role for 5-HT in modulating startle plasticity and increase our understanding of the neural and molecular basis of a trade-off between reproduction and predation. [ABSTRACT FROM AUTHOR]

The article discusses whether simple networks can implement flexible and complex decisions similarly. It says that decision-making is a basic aspect of life on all levels. According to the author, simpler systems can combine accessibility with complexity at the cellular level. An overview of the neuronal decision-making in the cortex and the evidence of how saccadic decision-making produces a good model for understanding flexible and complex decision-making are also presented.

The Roundabout (Robo) family of receptors and their Slit ligands play well-established roles in axonal guidance, including in humans where horizontal gaze palsy with progressive scoliosis (HGPPS) is caused by mutations in the robo3 gene. Although significant progress has been made toward understanding the mechanism by which Robo receptors establish commissural projections in the central nervous system, less is known about how these projections contribute to neural circuits mediating behavior. In this study, we report cloning of the zebrafish behavioral mutant twitch twice and show that twitch twice encodes robo3. We show that in mutant hindbrains the axons of an identified pair of neurons, the Mauthner cells, fail to cross the midline. The Mauthner neurons are essential for the startle response, and in twitch twice/ robo3 mutants misguidance of the Mauthner axons results in a unidirectional startle response. Moreover, we show that twitch twice mutants exhibit normal visual acuity but display defects in horizontal eye movements, suggesting a specific and critical role for twitch twice/ robo3 in sensory-guided behavior. [ABSTRACT FROM AUTHOR]

Studies of vertebrate brain evolution have focused primarily on patterns of gene expression or changes in size and organization of major brain regions. The Mauthner cell, an important reticulospinal neuron that functions in the startle response of many species, provides an opportunity for evolutionary comparisons at the cellular level. Despite broad interspecific similarities in Mauthner cell morphology, the motor patterns and startle behaviors it initiates vary markedly. Response diversity has been hypothesized to result, in part, from differences in the structure and function of the Mauthner cell-associated axon cap. We used light microscopy techniques to compare axon cap morphology across a wide range of species, including all four extant basal actinopterygian orders, representatives of a variety of teleost lineages and lungfishes, and we combined our data with published descriptions of axon cap structure. The ‘composite’ axon cap, observed in teleosts, is an organized conglomeration of glia and fibers of inhibitory and excitatory interneurons. Lungfish, amphibian tadpoles and several basal actinopterygian fishes have ‘simple’ axon caps that appear to lack glia and include few fibers. Several other basal actinopterygian fishes have ‘simple-dense’ caps that include greater numbers of fibers than simple caps, but lack the additional elements and organization of composite caps. Phylogenetic mapping shows that through evolution there are discrete transitions in axon cap morphology occurring at the base of gnathostomes, within basal actinopterygians, and at the base of the teleost radiation. Comparing axon cap evolution to the evolution of startle behavior and motor pattern provides insight into the relationship between Mauthner cell-associated structures and their functions in behavior. Copyright © 2009 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Brainstem reticulospinal neurons (RSNs) serve as the major descending system in vertebrate sensorimotor integration. One of the paired RSNs in zebrafish, the Mauthner (M) cell, is thought to initiate fast escape from sudden noxious stimuli. Two other paired RSNs, morphologically homologous to the M-cell, are also suggested to play key roles in controlling fast escape. However, the relationship among activities of the M-cell and its homologs during fast escape and the sensory inputs that elicit escape via their activation are unclear. We have monitored hindbrain RSN activity simultaneously with tail flip movement during fast escape in zebrafish. Confocal calcium imaging of RSNs was performed on larvae rostrally embedded in agar but with their tails allowed to move freely. Application of a pulsed waterjet to the otic vesicle (OV) to activate acousticovestibular input elicited contralateral fast tail flips with short latency and an apparent Ca2+ increase, reflecting a single action potential, in the ipsilateral M-cell (M-escape). Application of waterjet to head skin for tactile stimulation elicited fast escapes, but onset was delayed and the M-cell did not fire (non-M-escape). After eliminating either the M-cell or OV, only non-M-escape was initiated. Simultaneous high-speed confocal imaging of the M-cell and one of its homologs, MiD3cm, revealed complementary activation during fast escape: MiD3cm activity was low during M-escape but high during non-M-escape. These results suggest that M-cell firing is necessary for fast escape with short latency elicited by acousticovestibular input and that MiD3cm is more involved in non-M-escape driven by head-tactile input. [ABSTRACT FROM AUTHOR]

Although regulation of chemical transmission is known to involve the interaction of receptors with scaffold proteins, little is known about the existence of protein-protein interactions in regulating gap junction-mediated electrical synapses. The scaffold protein zonula-occludens-1 (ZO-1), a member of the MAGUK family of proteins was reported to interact with several connexins (Cxs). We show here that ZO-1 extensively colocalizes with Cx35 at identifiable "mixed" (electrical and chemical) contacts on goldfish Mauthner cells, a model synapse for the study of vertebrate electrical transmission where it is possible to correlate physiological properties with molecular composition. Further, our analysis indicates that these proteins directly interact at goldfish electrical synapses. In contrast to Cx43, which interacts with ZO-1 via the PDZ2 domain, Cx35 interacts with ZO-1 via the PDZ1 domain, and this association is of lower affinity. The properties of the ZO-1/Cx35 association suggest the existence of a more dynamic relation between these two proteins, possibly including a role of ZO-1 in regulating gap junctional conductance at these highly modifiable electrical synapses. The interaction of ZO-1 with conserved regions of the C termini of Cx35/Cx36 orthologs may have a common function at electrical synapses of mammals and other vertebrates. [ABSTRACT FROM AUTHOR]

Abstract: Protein kinase C isozymes are a biologically diverse group of enzymes known to be involved in a wide variety of cellular processes. They fall into three families (conventional, novel and atypical) depending upon their mode of activation. Several classes of zebrafish neurons have been shown to express PKCα during development, but the expression of other isoforms remains unknown. In this study we performed immunohistochemistry to determine if zebrafish express various isoforms of PKC. We used antibodies to test for the presence of enzymes that are thought to be preferentially expressed in the nervous system (PKCγ, βII, δ, ɛ, θ and ζ). Here, we show that PKCγ, ɛ, θ and ζ are expressed in the zebrafish CNS. Anti-PKCγ labels Rohon-Beard sensory neurons and Mauthner cells. PKCɛ and ζ staining is widespread in the CNS, and PKCθ and βII are expressed in skeletal muscle, especially at intersegmental boundaries. Immunoblot experiments confirm the specificity of the antibodies in zebrafish and indicate that the fish isoforms of PKCγ, βII, ɛ and ζ are similar to the mammalian isoforms. Interestingly, PKCθ appears to be similar to PKCθII, which, to date, has been found exclusively in mouse testis, but not in the mammalian CNS. Overall, our findings indicate that several different PKC isoforms are expressed in zebrafish, and that Rohon-Beard, Mauthner cells and muscle fibers preferentially express some isoforms over others. [Copyright &y& Elsevier]

When startled by sound, goldfish make large turns away from a rostral stimulus and small responses away from caudal stimuli, suggesting that rostral startling stimuli recruit larger pools of reticulospinal neurons in the Brainstem Escape Network (BEN) than do caudal stimuli. Consistent with this idea, the zebrafish Mauthner (M-) cell fires when the fish is startled by tail-directed stimuli, but the M-cell homologues (MiD2cm and MiD3cm) are also recruited when the fish is startled by displacing the head. Because vision is known to modulate M-cell activity, a nonstartling, modulatory sensory ‘signal’ conveyed to the reticular formation may be stronger if the visual sensory image is from a rostral vs. caudal spatial location and could account for a differential neuron pool recruitment and response magnitude. In this study, electrophysiological recordings from cichlid Mauthner neurons showed that visual stimulation of the caudal retina (by a rostral cue) generates a depolarization that is about 1.5 times the amplitude of that generated by stimulation of the rostral retina (by a caudal cue). In behavioral testing, where fish were stimulated visually for 30 ms and then startled by sound, fish startled in the presence of a rostral visual stimulus performed larger amplitude and faster turns than when startled in the presence of a caudal visual stimulus. Thus, M-cell potentials might reflect the strength of visual input to the BEN in general. For a particular visual spatial location, the relative strength of descending visual input appears to contribute to a recruitment of a reticulospinal neuron population that generates a turn magnitude appropriate to the visual cue, and suggests that a retinotopic representation is preserved in the BEN. Copyright © 2006 S. Karger AG, Basel [ABSTRACT FROM AUTHOR]

Although behavior is ultimately guided by decision-making neurons and their associated networks, the mechanisms underlying neural decision-making in a behaviorally relevant context remain mostly elusive. To address this question, we analyzed goldfish escapes in response to distinct visual looming stimuli with high-speed video and compared them with electrophysiological responses of the Mauthner cell (M-cell), the threshold detector that initiates such behaviors. These looming stimuli evoke powerful and fast body-bend (C-start) escapes with response probabilities between 0.7 and 0.91 and mean latencies ranging from 142 to 716 ms. Chronic recordings showed that these C-starts are correlated with M-cell activity. Analysis of response latency as a function of the different optical parameters characterizing the stimuli suggests response threshold is closely correlated to a dynamically scaled function of angular retinal image size, θ(t), specifically κ(t) = θ(t = δ x e -βθ(t - δ), where the exponential term progressively reduces the weight of θ(t). Intracellular recordings show that looming stimuli typically evoked bursts of graded EPSPs with peak amplitudes up to 9 mV in the M-cell. The proposed scaling function κ(t) predicts the slope of the depolarizing envelope of these EPSPs and the timing of the largest peak. An analysis of the firing rate of presynaptic inhibitory interneurons suggests the timing of the EPSP peak is shaped by an interaction of excitatory and inhibitory inputs to the M-cell and corresponds to the temporal window in which the probabilistic decision of whether or not to escape is reached. [ABSTRACT FROM AUTHOR]

Protein kinase C (PKC) is a family of enzymes involved in a wide range of biological functions. We investigated the expression of PKC-positive cells in zebrafish embryos and larvae within the first week of development to determine the developmental profile of PKC-containing cells. Our other goal was to determine if PKCα was associated with Rohon-Beard neurons during the first 5 days of development, when they are reported to undergo apoptosis. First, we confirmed the specificity of the antibodies by Western blotting zebrafish brain homogenates with anti-PKC and anti-PKCα, and detected single protein bands of approximately 78-82 kDa in size. Immunohistochemistry showed that several types of neurons were labeled, including neurons in the trigeminal ganglia, the dorsal spinal cord, and the dorsal root ganglia. Double-labeling with anti-PKCα and both anti-Islet-1 and zn12 confirmed the identity of the PKC-positive cells in the brain as trigeminal neurons, and in the spinal cord as Rohon-Beard cells. Some Rohon-Beard cells were labeled with anti-PKCα up to 7 days post fertilization (dpf). We performed TUNEL labeling and found no correlation between TUNEL-labeled and PKCα-labeled Rohon-Beard cells, suggesting that PKCα is not involved in Rohon-Beard apoptosis. Only approximately 40% of the ≈130 Rohon-Beard cells at 24 h postfertilization (hpf) were positively labeled for PKC. Mauthner cells were labeled by anti-PKC, but not anti-PKCα, suggesting that the major form of PKC within these cells was not PKCα. © 2004 Wiley Periodicals, Inc. J Neurobiol, 2005 [ABSTRACT FROM AUTHOR]

In the hindbrain of zebrafish and goldfish, reticulospinal (RS) neurons are arranged in seven segments, with segmental homologs in adjacent segments. The Mauthner cell (M-cell) in the fourth segment (r4) is known to trigger fast escape behavior. Its serial homologs, MiD2cm in r5 and MiD3cm in r6, are predicted to contribute to this behavior, which can be evoked by head-tap stimuli. However, little is known about their input- output properties. Therefore, we studied afferent projections from the auditory posterior eighth nerve (pVIIIn) and firing properties of MiD2cm and MiD3cm for comparison with the M-cell in adult goldfish. Labeling of RS neurons and the pVIIIn afferents with fluorescent tracers showed that the pVIIIn projected to r4 -r6. Tone burst and electrical stimulation of the pVIIIn evoked EPSPs in the M-cell, MiD2cm, and MiD3cm. Stepwise depolarization typically elicited a single spike at the onset in the M-cell but repetitive spiking in MiD2cm and MiD3cm. This atypical property of the M-cell was mediated by dendrotoxin-I (DTX-I)-sensitive voltage-gated potassium channels together with recurrent inhibition, because combined application of DTX-I, strychnine, and bicuculline led to continuous repetitive firing in M-cells. The M-cell but not MiD2cm or MiD3cm expressed Kv1.2, a DTX-I-sensitive potassium channel subunit. Thus, the M-cell and its segmental homologs may sense common auditory information but send different outputs to the spinal circuits to control adaptive escape behavior. [ABSTRACT FROM AUTHOR]

Hox genes are critical for establishing the segmental pattern of the vertebrate hindbrain. Changes in their expression can alter neural organization of hindbrain segments and may be a mechanism for brain evolution. To test the hypothesis that neurons induced through changes in Hox gene expression can integrate into functional neural circuits, we examined the roles of ectopic Mauthner cells (M-cells) in the escape response of larval zebrafish. The activity of the paired Mauthner cells in rhombomere 4 (r4) has been shown to be critical for generating a high-performance startle behavior in response to stimulation of the tail (Liu and Fetcho,1999). Previous studies have found that misexpression of particular Hox genes causes ectopic M-cells to be generated in r2 in addition to the r4 cells (Alexandra et al., 1996; McClintock et al., 2001). With calcium imaging, we found that the homeotically transformed neurons respond to startle stimuli. To determine the roles of ectopic and endogenous M-cells in the behavior, we lesioned the r2, r4, or both M-cells with cell-specific laser lesion and examined the effect on startle performance. Lesion of the normal M-cells did not decrease escape performance when the ectopic cells were present. These results indicate that the homeotically transformed Mauthner cells are fully functional in the escape circuit and are functionally redundant with normal M-cells. We suggest that such functional redundancy between neurons may provide a substrate for evolution of neural circuits. [ABSTRACT FROM AUTHOR]

To study the modulatory influences of visual information on Mauthner (M-) initiated C-start responses, short interval visual stimuli were presented to individual cichlid fish prior to being startled with a sound pulse. Because the axon of each Mauthner neuron activates trunk musculature contralateral to the soma, the initial direction of an ensuing startle response provides a behavioral measure of which cell has been driven closer to threshold by visual cues at the time the sound pulse causes one cell to fire. When an LED was illuminated on one side of the fish for 10 ms prior to a startling sound pulse, cichlids reliably turned toward the visual cue. At durations of 15 and 20 ms, fish turned away from the same stimulus. Thus, behavioral evidence suggests that the M-cell contralateral to a visual stimulus appears to be excited first but visual excitation of the M-cell ipsilateral to the visual stimulus follows and predominates. Consistent with the behavioral results, visually evoked excitatory potentials recorded intracellularly in the cichlid M-cells were complex, with initial PSPs showing latencies of about 11.6 ms from contralateral eye stimulation and 15.5 ms from stimulating the ipsilateral eye. PSP latencies in goldfish were longer and more similar for stimuli to the two eyes (about 22 ms). For contrast, sound-evoked PSPs begin within 2 ms. The relative long latencies for visually regulating M-cell function suggest that vision is most adaptive for biasing response direction prior to rather than during a predator's attack. [ABSTRACT FROM AUTHOR]

Stimulation of the spinal cord of the electric fish Gymnotus carapo, evoked an abrupt increase in the discharge rate of the electric organ. At the maximum of this response, the rate increased an average of 26 ± 11.8%. The duration of the response was 4.9 ± 2.12 s; its latency was 10.4 ± 1.1 ms. Activation of the Mauthner axon played a decisive role in this phenomenon as indicated by the following: (1) recordings from the axon cap of the Mauthner cell demonstrated that the response was evoked if the Mauthner axon was antidromically activated and (2) a response that was similar to that produced by spinal cord stimulation, was elicited by intracellular stimulation of either Mauthner cell. Stimulation of the eighth nerve could also increase the discharge rate of the electric organ. The effect was greater if a Mauthner cell action potential was elicited. The findings described in the present report, indicate the existence of a functional connection between the Mauthner cell and the electromotor system in Gymnotus carapo. This connection may function to enhance the electrolocative sampling of the environment during Mauthner-cell mediated behaviors. This is a novel function for the Mauthner cell. [ABSTRACT FROM AUTHOR]

Stimulation of the spinal cord of the electric fish Gymnotus carapo, evoked an abrupt increase in the discharge rate of the electric organ. At the maximum of this response, the rate increased an average of 26 ± 11.8%. The duration of the response was 4.9 ± 2.12 s; its latency was 10.4 ± 1.1 ms. Activation of the Mauthner axon played a decisive role in this phenomenon as indicated by the following: (1) recordings from the axon cap of the Mauthner cell demonstrated that the response was evoked if the Mauthner axon was antidromically activated and (2) a response that was similar to that produced by spinal cord stimulation, was elicited by intracellular stimulation of either Mauthner cell. Stimulation of the eighth nerve could also increase the discharge rate of the electric organ. The effect was greater if a Mauthner cell action potential was elicited. The findings described in the present report, indicate the existence of a functional connection between the Mauthner cell and the electromotor system in Gymnotus carapo. This connection may function to enhance the electrolocative sampling of the environment during Mauthner-cell mediated behaviors. This is a novel function for the Mauthner cell. [ABSTRACT FROM AUTHOR]

Once initiated, rapid escape responses of teleost fishes are thought to be completed without additional sensory modification. This suggests that the motor program for a particular response is selected for by the constellation of sensory cues existing at the time of the releasing stimulus. This paper presents initial evidence that a highly specialized, phylogenetically recent electrosensory system is integrated with a primitive motor system and allows an animal to continuously monitor its environment for producing accurate escape behaviors. Behavioral testing for directed startle responses in a 'Y-maze' demonstrates that when presented immediately before an acoustic startle stimulus, electric fish ( Eigenmannia virescens), direct their response away from the cue (a transient shorting of their electric field). Thus, electrosensory cues as brief as 100 ms provide directional information to the escape motor network. In electric fish that are curarized to facilitate intracellular recording, the normal electric organ discharge is attenuated. When an electronically generated replacement field of the same frequency and amplitude as the fish's normal signal is shorted, a fast-rising, 7 ms latency post-synaptic potential is evoked from the Mauthner cell. Similar PSPs are generated by turning the replacement stimulus on and off. In some recordings, removing the S1 replacement field elicits a rebound of other afferent activity to the Mauthner cell; replacing the field suppresses this activity. [ABSTRACT FROM AUTHOR]

The Mauthner (M-) cells, a bilateral pair of medullary neurons in fish, initiate the characteristic 'C-start' predatory escape response of teleosts. Similar movements have been described during hatching, social interactions, and feeding. M-cell firing, however, has not been correlated directly with these other behaviors. The objective of this study was to determine whether the M-cell, in addition to escape, plays a role in feeding. [ABSTRACT FROM AUTHOR]

A comparative anatomical and behavioral approach was used to elucidate the role of medullary networks associated with Mauthner cells (M-cells) in determining the type of fast startle responses of elongate bony fish with an anguilliform type of locomotion. The M-cell initiates fast startle responses in goldfish and is thought to initiate such responses in other fish as well [Wilson, 1959; Eaton et al., 1977; Currie and Carlsen, 1987; Currie, 1991]. Goldfish M-cells have a specialized structure, termed an axon cap, that surrounds their axon hillock-initial segment region and is critical for the control of M-cell excitability. The M-cell axon cap is present in some elongate fish such as the African lungfish (Protopterus, Lepidosireniformes) and absent in others such as the American eel (Anguilla, Anguilliformes). The lungfish startle response is characterized by a rotation of the head and upper trunk, similar to the initial phase of the goldfish (Carassius, Cypriniformes) startle response. The American eel, on the other hand, displays a withdrawal-type startle response. We hypothesize that the withdrawal-type startle response of the American eel results from the absence of inhibitory networks associated with the M-cell axon cap. The correlation of the absence of an axon cap and a withdrawal-type startle response may be a general feature of all Anguilliformes and elongate ray-finned fish with an anguilliform type of locomotion. [ABSTRACT FROM AUTHOR]

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The Mauthner cell can first be recognized in Xenopus embryos at stage 31-32 by its position and by the size of its nucleus and nucleolus. At early stages the perikaryon is dominated by a large ellipsoidal nucleus containing a single relatively compact nucleolus. The cytoplasm is characterized by numerous free ribosomes, poorly developed membrane systems, yolk platelets and globules, and by the presence of dense bodies containing what appear to be remnants of yolk breakdown. During subsequent development, membranes of both the endoplasmic reticulum and the Golgi apparatus become increasingly numerous. Long sinuous tubules of the endoplasmic reticulum are replaced by shorter repeatedly branching cisterns, and incidents of fusion between these elements and the nuclear envelope are less frequent. Large intramitochondrial granules and yolk elements are seldom encountered. The number of multivesicular bodies, alveolate vesicles and small secondary lysosomes increases. The originally nearly homogeneous cytoplasm is rearranged into areas rich in ribosomes and membranous organelles separated by channels containing predominantly neurofilaments and microtubules. At intermediate stages of development a cytoplasmic inclusion consisting of short arcs of parallel beaded strands, each 130-150 Å in diameter, is often present. The relationship of these morphological changes to the overall synthetic pattern of the cell is discussed. It is suggested that they signal the end of 'seeking' growth by the cell. [ABSTRACT FROM AUTHOR]

The article discusses about the importance of dendritic structure for neuronal computation and neuronal functioning. Topics discussed include integration of combined sound and optic tectum stimulation on the dendrites of Mauthner cells; response of dendrites to the auditory channel as well as visual channel; and visual and auditory stimulation led to complex voltage traces.