Object approach computation by a giant neuron and its relation with the speed of escape in the crab Neohelice

Upon detection of an approaching object, the crab Neohelice granulata continuously regulates the direction and speed of escape according to ongoing visual information. These visuomotor transformations are thought to be largely accounted for by a small number of motion-sensitive giant neurons projecting from the lobula (third optic neuropil) towards the supraesophageal ganglion. One of these elements, the monostratified lobula giant neuron of type 2 (MLG2), proved to be highly sensitive to looming stimuli (a 2D representation of an object approach). By performing in vivo intracellular recordings, we assessed the response of the MLG2 neuron to a variety of looming stimuli representing objects of different sizes and velocities of approach. This allowed us to: (1) identify some of the physiological mechanisms involved in the regulation of the MLG2 activity and test a simplified biophysical model of its response to looming stimuli; (2) identify the stimulus optical parameters encoded by the MLG2 and formulate a phenomenological model able to predict the temporal course of the neural firing responses to all looming stimuli; and (3) incorporate the MLG2-encoded information of the stimulus (in terms of firing rate) into a mathematical model able to fit the speed of the escape run of the animal. The agreement between the model predictions and the actual escape speed measured on a treadmill for all tested stimuli strengthens our interpretation of the computations performed by the MLG2 and of the involvement of this neuron in the regulation of the animal's speed of run while escaping from objects approaching with constant speed.Fil: Oliva, Damian Ernesto. Universidad Nacional de Quilmes. Departamento de Ciencia y Tecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Differences in the escape response of a grapsid crab in the field and in the laboratory

Escape behaviours of prey animals are frequently used to study the neural control of behaviour. Escape responses are robust, fast, and can be reliably evoked under both field and laboratory conditions. Many escape responses are not as simple as previously suggested, however, and are often modulated by a range of contextual factors. To date it has been unclear to what extent behaviours studied in controlled laboratory experiments are actually representative of the behaviours that occur under more natural conditions. Here we have used the model species, Neohelice granulata, a grapsid crab, to show that there are significant differences between the crabs' escape responses in the field compared to those previously documented in laboratory experiments. These differences are consistent with contextual adjustments such as the availability of a refuge and have clear consequences for understanding the crabs' neural control of behaviour. Furthermore, the methodology used in this study mirrors the methodology previously used in fiddler crab research, allowing us to show that the previously documented differences in escape responses between these grapsid species are real and substantial. Neohelice's responses are delayed and more controlled. Overall, the results highlight the adaptability and flexibility of escape behaviours and provide further evidence that the neural control of behaviour needs to be address in both the laboratory and field context.Fil: Hemmi, J. M.. University of Western Australia; AustraliaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Brain modularity in arthropods: individual neurons that support “what” but not “where” memories

Experiments with insects and crabs have demonstrated their remarkable capacity to learn and memorize complex visual features (Giurfa et al., 2001; Pedreira and Maldonado, 2003; Chittka and Niven, 2009). Such abilities are thought to require modular brain processing similar to that occurring in vertebrates (Menzel and Giurfa, 2001). Yet, physiological evidence for this type of functioning in the small brains of arthropods is still scarce (Liu et al., 1999, 2006; Menzel and Giurfa, 2001). In the crab Chasmagnathus granulatus, the learning rate as well as the long-term memory of a visual stimulus has been found to be reflected in the performance of identified lobula giant neurons (LGs) (Tomsic et al., 2003). The memory can only be evoked in the training context, indicating that animals store two components ofthe learned experience, one relatedtothe visual stimulus and one relatedtothe visual context (Tomsic et al., 1998; Hermitte et al., 1999). By performing intracellular recordings inthe intact animal, we showthatthe ability of crabsto generalizethe learned stimulus into new space positions and to distinguish it from a similar but unlearned stimulus, two of the main attributes of stimulus memory, is reflected bythe performance ofthe LGs. Conversely, wefoundthat LGs do not supportthe visual context memory component. Our results provide physiological evidence that the memory traces regarding “what” and “where” are stored separately in the arthropod brain.Fil: Sztarker, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Predation risk modifies behaviour by shaping the response of identified brain neurons

Interpopulation comparisons in species that show behavioural variations associated with particular ecological disparities offer good opportunities for assessing how environmental factors may foster specific functional adaptations in the brain. Yet, studies on the neural substrate that can account for interpopulation behavioural adaptations are scarce. Predation is one of the strongest driving forces for behavioural evolvability and, consequently, for shaping structural and functional brain adaptations. We analysed the escape response of crabs Neohelice granulata from two isolated populations exposed to different risks of avian predation. Individuals from the high-risk area proved to be more reactive to visual danger stimuli (VDS) than those from an area where predators are rare. Control experiments indicate that the response difference was specific for impending visual threats. Subsequently, we analysed the response to VDS of a group of giant brain neurons that are thought to play a main role in the visually guided escape response of the crab. Neurons from animals of the population with the stronger escape response were more responsive to VDS than neurons from animals of the less reactive population. Our results suggest a robust linkage between the pressure imposed by the predation risk, the response of identified neurons and the behavioural outcome.Fil: Magani, Fiorella. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Luppi, Tomas Atilio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Nuñez, Jesus Dario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Marinas y Costeras. Universidad Nacional de Mar del Plata. Facultad de Ciencia Exactas y Naturales. Instituto de Investigaciones Marinas y Costeras; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Visuo-motor transformations involved in the escape response to looming stimuli in the crab Neohelice (=Chasmagnathus) granulata

Escape responses to directly approaching predators represent one instance of an animal s ability to avoid collision. Usually, such responses can be easily evoked in the laboratory using two-dimensional computer simulations of approaching objects, known as looming stimuli. Therefore, escape behaviors are considered useful models for the study of computations performed by the brain to efficiently transform visual information into organized motor patterns. The escape response of the crab Neohelice (previously Chasmagnathus) granulata offers an opportunity to investigate the processing of looming stimuli and its transformation into complex motor patterns. Here we studied the escape performance of this crab to a variety of different looming stimuli. The response always consisted of a vigorous run away from the stimulus. However, the moment at which it was initiated, as well as the developed speed, closely matched the expansion dynamics of each particular stimulus. Thus, we analyzed the response events as a function of several variables that could theoretically be used by the crab (angular size, angular velocity, etc.). Our main findings were that: (1) the decision to initiate the escape run is made when the stimulus angular size increases by 7deg; (2) the escape run is not a ballistic kind of response, as its speed is adjusted concurrently with changes in the optical stimulus variables; and (3) the speed of the escape run can be faithfully described by a phenomenological input–output relationship based on the stimulus angular increment and the angular velocity of the stimulus.Fil: Oliva, Damian Ernesto. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Unidirectional Optomotor Responses and Eye Dominance in Two Species of Crabs

Animals, from invertebrates to humans, stabilize the panoramic optic flow through compensatory movements of the eyes, the head or the whole body, a behavior known as optomotor response (OR). The same optic flow moved clockwise or anticlockwise elicits equivalent compensatory right or left turning movements, respectively. However, if stimulated monocularly, many animals show a unique effective direction of motion, i.e., a unidirectional OR. This phenomenon has been reported in various species from mammals to birds, reptiles, and amphibious, but among invertebrates, it has only been tested in flies, where the directional sensitivity is opposite to that found in vertebrates. Although OR has been extensively investigated in crabs, directional sensitivity has never been analyzed. Here, we present results of behavioral experiments aimed at exploring the directional sensitivity of the OR in two crab species belonging to different families: the varunid mud crab Neohelice granulata and the ocypode fiddler crab Uca uruguayensis. By using different conditions of visual perception (binocular, left or right monocular) and direction of flow field motion (clockwise, anticlockwise), we found in both species that in monocular conditions, OR is effectively displayed only with progressive (front-to-back) motion stimulation. Binocularly elicited responses were directional insensitive and significantly weaker than monocular responses. These results are coincident with those described in flies and suggest a commonality in the circuit underlying this behavior among arthropods. Additionally, we found the existence of a remarkable eye dominance for the OR, which is associated to the size of the larger claw. This is more evident in the fiddler crab where the difference between the two claws is huge.Fil: Barnatan, Yair Benjamín. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Sztarker, Julieta. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

Direction selective neurons responsive to horizontal motion in a crab reflect an adaptation to prevailing movements in flat environments

All animals need information about the direction of motion to be able to track the trajectory of a target (prey, predator, cospecific) or to control the course of navigation. This information is provided by direction selective (DS) neurons, which respond to images moving in a unique direction. DS neurons have been described in numerous species including many arthropods. In these animals, the majority of the studies have focused on DS neurons dedicated to processing the optic flow generated during navigation. In contrast, only a few studies were performed on DS neurons related to object motion processing. The crab Neohelice is an established experimental model for the study of neurons involved in visually- guided behaviors. Here, we describe in male crabs of this species a new group of DS neurons that are highly directionally selective to moving objects. The neurons were physiologically and morphologically characterized by intracellular recording and staining in the optic lobe of intact animals. Because of their arborization in the lobula complex, we called these cells lobula complex directional cells (LCDCs). LCDCs also arborize in a previously undescribed small neuropil of the lateral protocerebrum. LCDCs are responsive only to horizontal motion. This nicely fits in the behavioral adaptations of a crab inhabiting a flat, densely crowded environment, where most object motions are generated by neighboring crabs moving along the horizontal plane.Fil: Scarano, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Tomsic, Daniel. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Sztarker, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentin

Unidirectional Optomotor Responses and Eye Dominance in Two Species of Crabs

Animals, from invertebrates to humans, stabilize the panoramic optic flow through compensatory movements of the eyes, the head or the whole body, a behavior known as optomotor response (OR). The same optic flow moved clockwise or anticlockwise elicits equivalent compensatory right or left turning movements, respectively. However, if stimulated monocularly, many animals show a unique effective direction of motion, i.e., a unidirectional OR. This phenomenon has been reported in various species from mammals to birds, reptiles, and amphibious, but among invertebrates, it has only been tested in flies, where the directional sensitivity is opposite to that found in vertebrates. Although OR has been extensively investigated in crabs, directional sensitivity has never been analyzed. Here, we present results of behavioral experiments aimed at exploring the directional sensitivity of the OR in two crab species belonging to different families: the varunid mud crab Neohelice granulata and the ocypode fiddler crab Uca uruguayensis. By using different conditions of visual perception (binocular, left or right monocular) and direction of flow field motion (clockwise, anticlockwise), we found in both species that in monocular conditions, OR is effectively displayed only with progressive (front-to-back) motion stimulation. Binocularly elicited responses were directional insensitive and significantly weaker than monocular responses. These results are coincident with those described in flies and suggest a commonality in the circuit underlying this behavior among arthropods. Additionally, we found the existence of a remarkable eye dominance for the OR, which is associated to the size of the larger claw. This is more evident in the fiddler crab where the difference between the two claws is huge

Multielectrode Recordings From Identified Neurons Involved in Visually Elicited Escape Behavior

A major challenge in current neuroscience is to understand the concerted functioning of distinct neurons involved in a particular behavior. This goal first requires achieving an adequate characterization of the behavior as well as an identification of the key neuronal elements associated with that action. Such conditions have been considerably attained for the escape response to visual stimuli in the crab Neohelice. During the last two decades a combination of in vivo intracellular recordings and staining with behavioral experiments and modeling, led us to postulate that a microcircuit formed by four classes of identified lobula giant (LG) neurons operates as a decision-making node for several important visually-guided components of the crab’s escape behavior. However, these studies were done by recording LG neurons individually. To investigate the combined operations performed by the group of LG neurons, we began to use multielectrode recordings. Here we describe the methodology and show results of simultaneously recorded activity from different lobula elements. The different LG classes can be distinguished by their differential responses to particular visual stimuli. By comparing the response profiles of extracellular recorded units with intracellular recorded responses to the same stimuli, two of the four LG classes could be faithfully recognized. Additionally, we recorded units with stimulus preferences different from those exhibited by the LG neurons. Among these, we found units sensitive to optic flow with marked directional preference. Units classified within a single group according to their response profiles exhibited similar spike waveforms and similar auto-correlograms, but which, on the other hand, differed from those of groups with different response profiles. Additionally, cross-correlograms revealed excitatory as well as inhibitory relationships between recognizable units. Thus, the extracellular multielectrode methodology allowed us to stably record from previously identified neurons as well as from undescribed elements of the brain of the crab. Moreover, simultaneous multiunit recording allowed beginning to disclose the connections between central elements of the visual circuits. This work provides an entry point into studying the neural networks underlying the control of visually guided behaviors in the crab brain.Fil: Cámera, Alejandro Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Belluscio, Mariano Andres. Universidad de Buenos Aires. Facultad de Medicina; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentin

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.Fil: Scarano, María Florencia. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Sztarker, Julieta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Medan, Violeta. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; ArgentinaFil: Berón de Astrada, Martín. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; ArgentinaFil: Tomsic, Daniel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Fisiología, Biología Molecular y Neurociencias. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Fisiología, Biología Molecular y Neurociencias; Argentina. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Departamento de Fisiología, Biología Molecular y Celular; Argentin