SUMMARY 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.

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.

Similar to most visual animals, crabs perform proper avoidance responses to objects directly approaching them. The monostratified lobula giant neurons of type 1 (MLG1) of crabs constitute an ensemble of 14-16 bilateral pairs of motion-detecting neurons projecting from the lobula (third optic neuropile) to the midbrain, with receptive fields that are distributed over the extensive visual field of the animal's eye. Considering the crab Neohelice (previously Chasmagnathus) granulata, here we describe the response of these neurons to looming stimuli that simulate objects approaching the animal on a collision course. We found that the peak firing time of MLG1 acts as an angular threshold detector signaling, with a delay of δ = 35 ms, the time at which an object reaches a fixed angular threshold of 49°. Using in vivo intracellular recordings, we detected the existence of excitatory and inhibitory synaptic currents that shape the neural response. Other functional features identified in the MLG1 neurons were phasic responses at the beginning of the approach, a relation between the stimulus angular velocity and the excitation delay, and a mapping between membrane potential and firing frequency. Using this information, we propose a biophysical model of the mechanisms that regulate the encoding of looming stimuli. Furthermore, we found that the parameter encoded by the MLG1 firing frequency during the approach is the stimulus angular velocity. The proposed model fits the experimental results and predicts the neural response to a qualitatively different stimulus. Based on these and previous results, we propose that the MLG1 neuron system acts as a directional coding system for collision avoidance.