Temporal Code-Driven Stimulation: Definition and Application to Electric Fish Signaling.

Closed-loop activity-dependent stimulation is a powerful methodology to assess information processing in biological systems. In this context, the development of novel protocols, their implementation in bioinformatics toolboxes and their application to different description levels open up a wide range of possibilities in the study of biological systems. We developed a methodology for studying biological signals representing them as temporal sequences of binary events. A specific sequence of these events (code) is chosen to deliver a predefined stimulation in a closed-loop manner. The response to this code-driven stimulation can be used to characterize the system. This methodology was implemented in a real time toolbox and tested in the context of electric fish signaling. We show that while there are codes that evoke a response that cannot be distinguished from a control recording without stimulation, other codes evoke a characteristic distinct response. We also compare the code-driven response to open-loop stimulation. The discussed experiments validate the proposed methodology and the software toolbox.

Automated pulse discrimination of two freely-swimming weakly electric fish and analysis of their electrical behavior during dominance contest.

Electric fishes modulate their electric organ discharges with a remarkable variability. Some patterns can be easily identified, such as pulse rate changes, offs and chirps, which are often associated with important behavioral contexts, including aggression, hiding and mating. However, these behaviors are only observed when at least two fish are freely interacting. Although their electrical pulses can be easily recorded by non-invasive techniques, discriminating the emitter of each pulse is challenging when physically similar fish are allowed to freely move and interact. Here we optimized a custom-made software recently designed to identify the emitter of pulses by using automated chirp detection, adaptive threshold for pulse detection and slightly changing how the recorded signals are integrated. With these optimizations, we performed a quantitative analysis of the statistical changes throughout the dominance contest with respect to Inter Pulse Intervals, Chirps and Offs dyads of freely moving Gymnotus carapo. In all dyads, chirps were signatures of subsequent submission, even when they occurred early in the contest. Although offs were observed in both dominant and submissive fish, they were substantially more frequent in submissive individuals, in agreement with the idea from previous studies that offs are electric cues of submission. In general, after the dominance is established the submissive fish significantly changes its average pulse rate, while the pulse rate of the dominant remained unchanged. Additionally, no chirps or offs were observed when two fish were manually kept in direct physical contact, suggesting that these electric behaviors are not automatic responses to physical contact.

Automatic realistic real time stimulation/recording in weakly electric fish: long time behavior characterization in freely swimming fish and stimuli discrimination.

Weakly electric fish are unique model systems in neuroethology, that allow experimentalists to non-invasively, access, central nervous system generated spatio-temporal electric patterns of pulses with roles in at least 2 complex and incompletely understood abilities: electrocommunication and electrolocation. Pulse-type electric fish alter their inter pulse intervals (IPIs) according to different behavioral contexts as aggression, hiding and mating. Nevertheless, only a few behavioral studies comparing the influence of different stimuli IPIs in the fish electric response have been conducted. We developed an apparatus that allows real time automatic realistic stimulation and simultaneous recording of electric pulses in freely moving Gymnotus carapo for several days. We detected and recorded pulse timestamps independently of the fish's position for days. A stimulus fish was mimicked by a dipole electrode that reproduced the voltage time series of real conspecific according to previously recorded timestamp sequences. We characterized fish behavior and the eletrocommunication in 2 conditions: stimulated by IPIs pre-recorded from other fish and random IPI ones. All stimuli pulses had the exact Gymontus carapo waveform. All fish presented a surprisingly long transient exploratory behavior (more than 8 h) when exposed to a new environment in the absence of electrical stimuli. Further, we also show that fish are able to discriminate between real and random stimuli distributions by changing several characteristics of their IPI distribution.

Single synapse information coding in intraburst spike patterns of central pattern generator motor neurons.

Burst firing is ubiquitous in nervous systems and has been intensively studied in central pattern generators (CPGs). Previous works have described subtle intraburst spike patterns (IBSPs) that, despite being traditionally neglected for their lack of relation to CPG motor function, were shown to be cell-type specific and sensitive to CPG connectivity. Here we address this matter by investigating how a bursting motor neuron expresses information about other neurons in the network. We performed experiments on the crustacean stomatogastric pyloric CPG, both in control conditions and interacting in real-time with computer model neurons. The sensitivity of postsynaptic to presynaptic IBSPs was inferred by computing their average mutual information along each neuron burst. We found that details of input patterns are nonlinearly and inhomogeneously coded through a single synapse into the fine IBSPs structure of the postsynaptic neuron following burst. In this way, motor neurons are able to use different time scales to convey two types of information simultaneously: muscle contraction (related to bursting rhythm) and the behavior of other CPG neurons (at a much shorter timescale by using IBSPs as information carriers). Moreover, the analysis revealed that the coding mechanism described takes part in a previously unsuspected information pathway from a CPG motor neuron to a nerve that projects to sensory brain areas, thus providing evidence of the general physiological role of information coding through IBSPs in the regulation of neuronal firing patterns in remote circuits by the CNS.

Connection topology selection in central pattern generators by maximizing the gain of information.

A study of a general central pattern generator (CPG) is carried out by means of a measure of the gain of information between the number of available topology configurations and the output rhythmic activity. The neurons of the CPG are chaotic Hindmarsh-Rose models that cooperate dynamically to generate either chaotic or regular spatiotemporal patterns. These model neurons are implemented by computer simulations and electronic circuits. Out of a random pool of input configurations, a small subset of them maximizes the gain of information. Two important characteristics of this subset are emphasized: (1) the most regular output activities are chosen, and (2) none of the selected input configurations are networks with open topology. These two principles are observed in living CPGs as well as in model CPGs that are the most efficient in controlling mechanical tasks, and they are evidence that the information-theoretical analysis can be an invaluable tool in searching for general properties of CPGs.

StdpC: a modern dynamic clamp.

With the advancement of computer technology many novel uses of dynamic clamp have become possible. We have added new features to our dynamic clamp software StdpC ("Spike timing-dependent plasticity Clamp") allowing such new applications while conserving the ease of use and installation of the popular earlier Dynclamp 2/4 package. Here, we introduce the new features of a waveform generator, freely programmable Hodgkin-Huxley conductances, learning synapses, graphic data displays, and a powerful scripting mechanism and discuss examples of experiments using these features. In the first example we built and 'voltage clamped' a conductance based model cell from a passive resistor-capacitor (RC) circuit using the dynamic clamp software to generate the voltage-dependent currents. In the second example we coupled our new spike generator through a burst detection/burst generation mechanism in a phase-dependent way to a neuron in a central pattern generator and dissected the subtle interaction between neurons, which seems to implement an information transfer through intraburst spike patterns. In the third example, making use of the new plasticity mechanism for simulated synapses, we analyzed the effect of spike timing-dependent plasticity (STDP) on synchronization revealing considerable enhancement of the entrainment of a post-synaptic neuron by a periodic spike train. These examples illustrate that with modern dynamic clamp software like StdpC, the dynamic clamp has developed beyond the mere introduction of artificial synapses or ionic conductances into neurons to a universal research tool, which might well become a standard instrument of modern electrophysiology.

A network of electronic neural oscillators reproduces the dynamics of the periodically forced pyloric pacemaker group.

Low-dimensional oscillators are a valuable model for the neuronal activity of isolated neurons. When coupled, the self-sustained oscillations of individual free oscillators are replaced by a collective network dynamics. Here, dynamical features of such a network, consisting of three electronic implementations of the Hindmarsh-Rose mathematical model of bursting neurons, are compared to those of a biological neural motor system, specifically the pyloric CPG of the crustacean stomatogastric nervous system. We demonstrate that the network of electronic neurons exhibits realistic synchronized bursting behavior comparable to the biological system. Dynamical properties were analyzed by injecting sinusoidal currents into one of the oscillators. The temporal bursting structure of the electronic neurons in response to periodic stimulation is shown to bear a remarkable resemblance to that observed in the corresponding biological network. These findings provide strong evidence that coupled nonlinear oscillators realistically reproduce the network dynamics experimentally observed in assemblies of several neurons.

Whole cell stochastic model reproduces the irregularities found in the membrane potential of bursting neurons.

Irregular intrinsic behavior of neurons seems ubiquitous in the nervous system. Even in circuits specialized to provide periodic and reliable patterns to control the repetitive activity of muscles, such as the pyloric central pattern generator (CPG) of the crustacean stomatogastric ganglion (STG), many bursting motor neurons present irregular activity when deprived from synaptic inputs. Moreover, many authors attribute to these irregularities the role of providing flexibility and adaptation capabilities to oscillatory neural networks such as CPGs. These irregular behaviors, related to nonlinear and chaotic properties of the cells, pose serious challenges to developing deterministic Hodgkin-Huxley-type (HH-type) conductance models. Only a few deterministic HH-type models based on experimental conductance values were able to show such nonlinear properties, but most of these models are based on slow oscillatory dynamics of the cytosolic calcium concentration that were never found experimentally in STG neurons. Based on an up-to-date single-compartment deterministic HH-type model of a STG neuron, we developed a stochastic HH-type model based on the microscopic Markovian states that an ion channel can achieve. We used tools from nonlinear analysis to show that the stochastic model is able to express the same kind of irregularities, sensitivity to initial conditions, and low dimensional dynamics found in the neurons isolated from the STG. Without including any nonrealistic dynamics in our whole cell stochastic model, we show that the nontrivial dynamics of the membrane potential naturally emerge from the interplay between the microscopic probabilistic character of the ion channels and the nonlinear interactions among these elements. Moreover, the experimental irregular behavior is reproduced by the stochastic model for the same parameters for which the membrane potential of the original deterministic model exhibits periodic oscillations.

Synaptic modulation of the interspike interval signatures of bursting pyloric neurons.

The pyloric network of the lobster stomatogastric nervous system is one of the best described assemblies of oscillatory neurons producing bursts of action potentials. While the temporal patterns of bursts have been investigated in detail, those of spikes have received less attention. Here we analyze the intraburst firing patterns of pyloric neurons and the synaptic interactions shaping their dynamics in millisecond time scales not performed before. We find that different pyloric neurons express characteristic, cell-specific firing patterns in their bursts. Nonlinear analysis of the interspike intervals (ISIs) reveals distinctive temporal structures ('interspike interval signatures'), which are found to depend on the synaptic connectivity of the network. We compare ISI patterns of the pyloric dilator (PD), lateral pyloric (LP), and ventricular dilator (VD) neurons in 1) normal conditions, 2) after blocking glutamatergic synaptic connections, and 3) in various functional configurations of the three neurons. Manipulation of the synaptic connectivity results in characteristic changes in the ISI signatures of the postsynaptic neurons. The intraburst firing pattern of the PD neuron is regularized by the inhibitory synaptic connection from the LP neuron as revealed in current-clamp experiments and also as reconstructed with a dynamic clamp. On the other hand, mutual inhibition between the LP and VD neurons tend to produce more irregular bursts with increased spike jitter. The results show that synaptic interactions fine-tune the output of pyloric neurons. The present data also suggest a way of processing of synaptic information: bursting neurons are capable of encoding incoming signals by altering the fine structure of their intraburst spike patterns.