Freezing of movements and its correspondence with MLG1 neurons response to looming stimuli in the crab Neohelice.

Upon visually detecting a moving predator animals often freeze, i.e. stop moving, to minimize being uncovered and to gather detailed information of the object's movements and properties. In certain conditions the freezing behavior can be enough to avoid a predatory menace but, when the risk is high or increases to a higher level, animals switch strategy and engage in an escape response. The neural bases underlying escape responses to visual stimuli are extensively investigated both in vertebrates and arthropods. However, those involved in freezing behaviors are much less studied. Here, we investigated the freezing behavior displayed by the crab Neohelice granulata when confronted with a variety of looming stimuli simulating objects of distinct sizes approaching on a collision course at different speeds. The experiments were performed in a treadmill-like device. Animals engaged in exploratory walks respond to the looming stimulus with freezing followed by escaping. The analysis of the stimulus optical variables shows that regardless of the looming dynamic, the freezing decision is made when the angular size of the object increases by 1.4°. In vivo intracellular recording responses of Monostratified Lobula Giant Neurons (MLG1) to the same looming stimuli show that the freezing times correlate with the times predicted by a hypothetical spike counter of this neuron.

Matched function of the neuropil processing optic flow in flies and crabs: the lobula plate mediates optomotor responses in Neohelice granulata.

When an animal rotates (whether it is an arthropod, a fish, a bird or a human) a drift of the visual panorama occurs over its retina, termed optic flow. The image is stabilized by compensatory behaviours (driven by the movement of the eyes, head or the whole body depending on the animal) collectively termed optomotor responses. The dipteran lobula plate has been consistently linked with optic flow processing and the control of optomotor responses. Crabs have a neuropil similarly located and interconnected in the optic lobes, therefore referred to as a lobula plate too. Here we show that the crabs' lobula plate is required for normal optomotor responses since the response was lost or severely impaired in animals whose lobula plate had been lesioned. The effect was behaviour-specific, since avoidance responses to approaching visual stimuli were not affected. Crabs require simpler optic flow processing than flies (because they move slower and in two-dimensional instead of three-dimensional space), consequently their lobula plates are relatively smaller. Nonetheless, they perform the same essential role in the visual control of behaviour. Our findings add a fundamental piece to the current debate on the evolutionary relationship between the lobula plates of insects and crustaceans.

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.