Audiovisual integration in the Mauthner cell enhances escape probability and reduces response latency.

Fast and accurate threat detection is critical for animal survival. Reducing perceptual ambiguity by integrating multiple sources of sensory information can enhance perception and reduce response latency. However, studies addressing the link between behavioral correlates of multisensory integration and its underlying neural basis are rare. Fish that detect an urgent threat escape with an explosive behavior known as C-start. The C-start is driven by an identified neural circuit centered on the Mauthner cell, an identified neuron capable of triggering escapes in response to visual and auditory stimuli. Here we demonstrate that goldfish can integrate visual looms and brief auditory stimuli to increase C-start probability. This multisensory enhancement is inversely correlated to the salience of the stimuli, with weaker auditory cues producing a proportionally stronger multisensory effect. We also show that multisensory stimuli reduced C-start response latency, with most escapes locked to the presentation of the auditory cue. We make a direct link between behavioral data and its underlying neural mechanism by reproducing the behavioral data with an integrate-and-fire computational model of the Mauthner cell. This model of the Mauthner cell circuit suggests that excitatory inputs integrated at the soma are key elements in multisensory decision making during fast C-start escapes. This provides a simple but powerful mechanism to enhance threat detection and survival.

Stimulus Contrast Information Modulates Sensorimotor Decision Making in Goldfish.

Animal survival relies on environmental information gathered by their sensory systems. We found that contrast information of a looming stimulus biases the type of defensive behavior that goldfish (Carassius auratus) perform. Low-contrast looms only evoke subtle alarm reactions whose probability is independent of contrast. As looming contrast increases, the probability of eliciting a fast escape maneuver, the C-start response, increases dramatically. Contrast information also modulates the decision of when to escape. Although response latency is known to depend on looming retinal size, we found that contrast acts as an additional parameter influencing this decision. When presenting progressively higher contrast stimuli, animals need shorter periods of stimulus processing to initiate the response. Our results comply with the notion that the decision to escape is a flexible process initiated with stimulus detection and followed by assessment of the perceived risk posed by the stimulus. Highly disruptive behaviors as the C-start are only observed when a multifactorial threshold that includes stimulus contrast is surpassed.

Optimal searching behaviour generated intrinsically by the central pattern generator for locomotion.

Efficient searching for resources such as food by animals is key to their survival. It has been proposed that diverse animals from insects to sharks and humans adopt searching patterns that resemble a simple Lévy random walk, which is theoretically optimal for 'blind foragers' to locate sparse, patchy resources. To test if such patterns are generated intrinsically, or arise via environmental interactions, we tracked free-moving Drosophila larvae with (and without) blocked synaptic activity in the brain, suboesophageal ganglion (SOG) and sensory neurons. In brain-blocked larvae, we found that extended substrate exploration emerges as multi-scale movement paths similar to truncated Lévy walks. Strikingly, power-law exponents of brain/SOG/sensory-blocked larvae averaged 1.96, close to a theoretical optimum (µ ≅ 2.0) for locating sparse resources. Thus, efficient spatial exploration can emerge from autonomous patterns in neural activity. Our results provide the strongest evidence so far for the intrinsic generation of Lévy-like movement patterns.

Binocular Neuronal Processing of Object Motion in an Arthropod.

Animals use binocular information to guide many behaviors. In highly visual arthropods, complex binocular computations involved in processing panoramic optic flow generated during self-motion occur in the optic neuropils. However, the extent to which binocular processing of object motion occurs in these neuropils remains unknown. We investigated this in a crab, where the distance between the eyes and the extensive overlapping of their visual fields advocate for the use of binocular processing. By performing in vivo intracellular recordings from the lobula (third optic neuropil) of male crabs, we assessed responses of object-motion-sensitive neurons to ipsilateral or contralateral moving objects under binocular and monocular conditions. Most recorded neurons responded to stimuli seen independently with either eye, proving that each lobula receives profuse visual information from both eyes. The contribution of each eye to the binocular response varies among neurons, from those receiving comparable inputs from both eyes to those with mainly ipsilateral or contralateral components, some including contralateral inhibition. Electrophysiological profiles indicated that a similar number of neurons were recorded from their input or their output side. In monocular conditions, the first group showed shorter response delays to ipsilateral than to contralateral stimulation, whereas the second group showed the opposite. These results fit well with neurons conveying centripetal and centrifugal information from and toward the lobula, respectively. Intracellular and massive stainings provided anatomical support for this and for direct connections between the two lobulae, but simultaneous recordings failed to reveal such connections. Simplified model circuits of interocular connections are discussed.SIGNIFICANCE STATEMENT Most active animals became equipped with two eyes, which contributes to functions like depth perception, objects spatial location, and motion processing, all used for guiding behaviors. In visually active arthropods, binocular neural processing of the panoramic optic flow generated during self-motion happens already in the optic neuropils. However, whether binocular processing of single-object motion occurs in these neuropils remained unknown. We investigated this in a crab, where motion-sensitive neurons from the lobula can be recorded in the intact animal. Here we demonstrate that different classes of neurons from the lobula compute binocular information. Our results provide new insight into where and how the visual information acquired by the two eyes is first combined in the brain of an arthropod.

Differential processing in modality-specific Mauthner cell dendrites.

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.

A network of visual motion-sensitive neurons for computing object position in an arthropod.

Highly active insects and crabs depend on visual motion information for detecting and tracking mates, prey, or predators, for which they require directional control systems containing internal maps of visual space. A neural map formed by large, motion-sensitive neurons implicated in processing panoramic flow is known to exist in an optic ganglion of the fly. However, an equivalent map for processing spatial positions of single objects has not been hitherto identified in any arthropod. Crabs can escape directly away from a visual threat wherever the stimulus is located in the 360° field of view. When tested in a walking simulator, the crab Neohelice granulata immediately adjusts its running direction after changes in the position of the visual danger stimulus smaller than 1°. Combining mass and single-cell staining with in vivo intracellular recording, we show that a particular class of motion-sensitive neurons of the crab's lobula that project to the midbrain, the monostratified lobula giants type 1 (MLG1), form a system of 16 retinotopically organized elements that map the 360° azimuthal space. The preference of these neurons for horizontally moving objects conforms the visual ecology of the crab's mudflat world. With a mean receptive field of 118°, MLG1s have a large superposition among neighboring elements. Our results suggest that the MLG1 system conveys information on object position as a population vector. Such computational code can enable the accurate directional control observed in the visually guided behaviors of crabs.

The Mauthner-cell circuit of fish as a model system for startle plasticity.

The Mauthner-cell (M-cell) system of teleost fish has a long history as an experimental model for addressing a wide range of neurobiological questions. Principles derived from studies on this system have contributed significantly to our understanding at multiple levels, from mechanisms of synaptic transmission and synaptic plasticity to the concepts of a decision neuron that initiates key aspects of the startle behavior. Here we will review recent work that focuses on the neurophysiological and neuropharmacological basis for modifications in the M-cell circuit. After summarizing the main excitatory and inhibitory inputs to the M-cell, we review experiments showing startle response modulation by temperature, social status, and sensory filtering. Although very different in nature, actions of these three sources of modulation converge in the M-cell network. Mechanisms of modulation include altering the excitability of the M-cell itself as well as changes in excitatory and inhibitor drive, highlighting the role of balanced excitation and inhibition for escape decisions. One of the most extensively studied forms of startle plasticity in vertebrates is prepulse inhibition (PPI), a sensorimotor gating phenomenon, which is impaired in several information processing disorders. Finally, we review recent work in the M-cell system which focuses on the cellular mechanisms of PPI and its modulation by serotonin and dopamine.

The 5-HT5A receptor regulates excitability in the auditory startle circuit: functional implications for sensorimotor gating.

Here we applied behavioral testing, pharmacology, and in vivo electrophysiology to determine the function of the serotonin 5-HT5A receptor in goldfish startle plasticity and sensorimotor gating. In an initial series of behavioral experiments, we characterized the effects of a selective 5-HT5A antagonist, SB-699551 (3-cyclopentyl-N-[2-(dimethylamino)ethyl]-N-[(4'-{[(2-phenylethyl)amino]methyl}-4-biphenylyl)methyl]propanamide dihydrochloride), on prepulse inhibition of the acoustic startle response. Those experiments showed a dose-dependent decline in startle rates in prepulse conditions. Subsequent behavioral experiments showed that SB-699551 also reduced baseline startle rates (i.e., without prepulse). To determine the cellular mechanisms underlying these behaviors, we tested the effects of two distinct selective 5-HT5A antagonists, SB-699551 and A-843277 (N-(2,6-dimethoxybenzyl)-N'[4-(4-fluorophenyl)thiazol-2-yl]guanidine), on the intrinsic membrane properties and synaptic sound response of the Mauthner cell (M-cell), the decision-making neuron of the startle circuit. Auditory-evoked postsynaptic potentials recorded in the M-cell were similarly attenuated after treatment with either 5-HT5A antagonist (SB-699551, 26.41 ± 3.98% reduction; A-843277, 17.52 ± 6.24% reduction). This attenuation was produced by a tonic (intrinsic) reduction in M-cell input resistance, likely mediated by a Cl(-) conductance, that added to the extrinsic inhibition produced by an auditory prepulse. Interestingly, the effector mechanisms underlying neural prepulse inhibition itself were unaffected by antagonist treatment. In summary, these results provide an in vivo electrophysiological characterization of the 5-HT5A receptor and its behavioral relevance and provide a new perspective on the interaction of intrinsic and extrinsic modulatory mechanisms in startle plasticity and sensorimotor gating.

Regionalization in the eye of the grapsid crab Neohelice granulata (=Chasmagnathus granulatus): variation of resolution and facet diameters.

Crabs have panoramic compound eyes, which can show marked regional specializations of visual acuity. These specializations are thought to be related to the particular features of the animal's ecological environment. Modern knowledge on the neuroanatomy and neurophysiology of the crabs' visual system mainly derives from studies performed in the grapsid crab Neohelice granulata (=Chasmagnathus granulatus). However, the organization of the visual sampling elements across the eye surface of this animal had not yet been addressed. We analyzed the sampling resolution across the eye of Neohelice by measuring the pseudopupil displacement with a goniometer. In addition, we measured the facet sizes in the different regions of the eye. We found that Neohelice possesses an acute band of high vertical resolution around the eye equator and an increase in horizontal sampling resolution and lenses diameter towards the lateral side of the eye. Therefore, the analysis of the optical apparatus indicates that this crab possesses greater visual acuity around the equator and at the lateral side of the eye. These specializations are compared with those found in different species of crabs and are discussed in connection to the particular ecological features of Neohelice's habitat.

Dopaminergic-induced changes in Mauthner cell excitability disrupt prepulse inhibition in the startle circuit of goldfish.

Prepulse inhibition (PPI) is a widespread sensorimotor gating phenomenon characterized by a decrease in startle magnitude if a nonstartling stimulus is presented 20-1,000 ms before a startling stimulus. Dopaminergic agonists disrupt behavioral PPI in various animal models. This provides an important neuropharmacological link to schizophrenia patients that typically show PPI deficits at distinct (60 ms) prepulse-pulse intervals. Here, we study time-dependent effects of dopaminergic modulation in the goldfish Mauthner cell (M-cell) startle network, which shows PPI-like behavioral and physiological startle attenuations. The unique experimental accessibility of the M-cell system allows investigating the underlying cellular mechanism with physiological stimuli in vivo. Our results show that the dopaminergic agonist apomorphine (2 mg/kg body wt) reduced synaptic M-cell PPI by 23.6% (n = 18; P = 0.009) for prepulse-pulse intervals of 50 ms, whereas other intervals showed no reduction. Consistently, application of the dopamine antagonist haloperidol (0.4 mg/kg body wt) restored PPI to control level. Current ramp injections while recording M-cell membrane potential revealed that apomorphine acts through a postsynaptic, time-dependent mechanism by deinactivating a M-cell membrane nonlinearity, effectively increasing input resistance close to threshold. This increase is most pronounced for prepulse-pulse intervals of 50 ms (47.9%, n = 8; P < 0.05) providing a time-dependent, cellular mechanism for dopaminergic disruption of PPI. These results provide, for the first time, direct evidence of dopaminergic modulation of PPI in the elementary startle circuit of vertebrates and reemphasize the potential of characterizing temporal aspects of PPI at the physiological level to understand its underlying mechanisms.

How visual space maps in the optic neuropils of a crab.

The Decapoda is the largest order of crustaceans, some 10,000 species having been described to date. The order includes shrimps, lobsters, crayfishes, and crabs. Most of these are highly visual animals that display complex visually guided behaviors and, consequently, large areas of their nervous systems are dedicated to visual processing. However, our knowledge of the organization and functioning of the visual nervous system of these animals is still limited. Beneath the retina lie three serially arranged optic neuropils connected by two chiasmata. Here, we apply dye tracers in different areas of the retina or the optic neuropils to investigate the organization of visual space maps in the optic neuropils of the brachyuran crab Chasmagnathus granulatus. Our results reveal the way in which the visual space is represented in the three main optic neuropils of a decapod. We show that the crabs' optic chiasmata are oriented perpendicular to each other, an arrangement that seems to be unique among malacostracans. Crabs use retinal position in azimuth and elevation to categorize visual stimuli; for instance, stimuli moving above or below the horizon are interpreted as predators or conspecifics, respectively. The retinotopic maps revealed in the present study create the possibility of relating particular regions of the optic neuropils with distinct behavioral responses elicited by stimuli occurring in different regions of the visual field.

Characterization of lobula giant neurons responsive to visual stimuli that elicit escape behaviors in the crab Chasmagnathus.

In the grapsid crab Chasmagnathus, a visual danger stimulus elicits a strong escape response that diminishes rapidly on stimulus repetition. This behavioral modification can persist for several days as a result of the formation of an associative memory. We have previously shown that a generic group of large motion-sensitive neurons from the lobula of the crab respond to visual stimuli and accurately reflect the escape performance. Additional evidence indicates that these neurons play a key role in visual memory and in the decision to initiate an escape. Although early studies recognized that the group of lobula giant (LG) neurons consisted of different classes of motion-sensitive cells, a distinction between these classes has been lacking. Here, we recorded in vivo the responses of individual LG neurons to a wide range of visual stimuli presented in different segments of the animal's visual field. Physiological characterizations were followed by intracellular dye injections, which permitted comparison of the functional and morphological features of each cell. All LG neurons consisted of large tangential arborizations in the lobula with axons projecting toward the midbrain. Functionally, these cells proved to be more sensitive to single objects than to flow field motion. Despite these commonalities, clear differences in morphology and physiology allowed us to identify four distinct classes of LG neurons. These results will permit analysis of the role of each neuronal type for visually guided behaviors and will allow us to address specific questions on the neuronal plasticity of LGs that underlie the well-recognized memory model of the crab.

Escape behavior and neuronal responses to looming stimuli in the crab Chasmagnathus granulatus (Decapoda: Grapsidae).

Behavioral responses to looming stimuli have been studied in many vertebrate and invertebrate species, but neurons sensitive to looming have been investigated in very few animals. In this paper we introduce a new experimental model using the crab Chasmagnathus granulatus, which allows investigation of the processes of looming detection and escape decision at both the behavioral and neuronal levels. By analyzing the escape response of the crab in a walking simulator device we show that: (i) a robust and reliable escape response can be elicited by computer-generated looming stimuli in all tested animals; (ii) parameters such as distance, speed, timing and directionality of the escape run, are easy to record and quantify precisely in the walking device; (iii) although the magnitude of escape varies between animals and stimulus presentations, the timing of the response is remarkably consistent and does not habituate at 3 min stimulus intervals. We then study the response of neurons from the brain of the crab by means of intracellular recordings in the intact animal and show that: (iv) two subclasses of previously identified movement detector neurons from the lobula (third optic neuropil) exhibit robust and reliable responses to the same looming stimuli that trigger the behavioral response; (v) the neurons respond to the object approach by increasing their rate of firing in a way that closely matches the dynamics of the image expansion. Finally, we compare the neuronal with the behavioral response showing that: (vi) differences in the neuronal responses to looming, receding or laterally moving stimuli closely reflect the behavioral differences to such stimuli; (vii) during looming, the crab starts to run soon after the looming-sensitive neurons begin to increase their firing rate. The increase in the running speed during stimulus approach faithfully follows the increment in the firing rate, until the moment of maximum stimulus expansion. Thereafter, the neurons abruptly stop firing and the animal immediately decelerates its run. The results are discussed in connection with studies of responses to looming stimuli in the locust.

Nectar foraging behaviour is affected by ant body size in Camponotus mus.

The nectivorous ant Camponotus mus shows a broad size variation within the worker caste. Large ants can ingest faster and larger loads than small ones. Differences in physiological abilities in fluid ingestion due to the insect size could be related to differences in decision-making according to ant size during nectar foraging. Sucrose solutions of different levels of sugar concentration (30% or 60%w/w), viscosity (high or low) or flow rate (ad libitum or 1microl/min) were offered in combination to analyse the behavioural responses to each of these properties separately. Differences were found depending on ant body size and the property compared. A regulated flow produced smaller crop loads for medium and large ants compared to the same solution given ad libitum. All foragers remained longer times feeding at the regulated flow source but larger ants often made longer interruptions. When sugar concentration was constant but viscosity was high, only large ants increased feeding time. Constant viscosity with different sugar concentration determined longer feeding time and bigger loads for the most concentrated solution for small but not for large ants. Small ants reached similar crop loads in a variety of conditions while large ants did not. These differences could be evidence of a possible specialization for nectar foraging based on ant body size.