Color-specific conditioning effects due to both orange and blue stimuli are observed in a Halobacterium salinarum strain devoid of putative methylatable sites on HtrI.

Behavioral responses of Halobacterium salinarum appear as changes in the frequency of motion reversals. Turning on orange light decreases the reversal frequency, whereas blue light induces reversals. Light pulses normally induce the same response as step-up stimuli. However, anomalous behavioral reactions, including inverse responses, are seen when stimuli are applied in sequence. The occurrence of a prior stimulus is conditioning for successive stimulation on a time scale of the same order of adaptational processes. These prolonged conditioning effects are color-specific. The only adaptation process identified so far is methylation of the transducers, and this could be somehow color-specific. Therefore we tested for the behavioral anomalies in a mutant in which all methylation sites on the transducer have been eliminated. The results show that behavioral anomalies are unaffected by the absence of methylation processes on the transducer.

Competition-integration of blue and orange stimuli in Halobacterium salinarum cannot occur solely in SRI photoreceptor.

Experiments on the integration of blue and orange stimuli in Halobacterium salinarum were performed by using different combinations of blue and orange steps. The results show that the prevalence of the blue stimulus over the orange one depends on both the blue and the orange light intensities. A quantitative analysis of the current hypotheses on the phototransduction of orange and UV-blue light stimuli is presented, showing that the balancing between the two antagonistic stimuli should depend only on the intensity of the blue stimulus and not on that of the orange one, provided that the combination of the two stimuli occurs linearly at the photoreceptor stage. We conclude that blue and orange stimuli elicit distinct intracellular signals whose integration occurs downstream of the photoreceptor.

Photoresponses of Halobacterium salinarum to repetitive pulse stimuli.

Halobacterium salinarum cells from 3-day-old cultures have been stimulated with different patterns of repetitive pulse stimuli. A short train of 0.6-s orange light pulses with a 4-s period resulted in reversal peaks of increasing intensity. The reverse occurred when blue light pulses were delivered as a finite train: with a 3-s period, the response declined in sequence from the first to the last pulse. To evaluate the response of the system under steady-state conditions of stimulation, continuous trains of pulses were also applied; whereas blue light always produced a sharply peaked response immediately after each pulse, orange pulses resulted in a declining peak of reversals that lasted until the subsequent pulse. An attempt to account for these results in terms of current excitation/adaptation models shows that additional mechanisms appear to be at work in this transduction chain.

Photosensory transduction in Halobacterium salinarium: evidence for a non-linear network of cross-talking pathways.

Transduction of light stimuli in Halobacterium salinarium is studied by behavioural experiments. Selected patterns of sequential stimuli (impinging on couples of the signalling states of its photoreceptors) show that a simple model integrating different stimuli is inadequate and that non linear interactions between different pathways occur through a network with several connections. The experiments reported herein yield rough but clear-cut information on the level of such interactions and shed new light on earlier findings.

Effects of sequential stimuli on Halobacterium salinarium photobehavior.

We analyzed the motor photoresponses of Halobacterium salinarium to different test stimuli applied after a first photophobic response produced by a step-down of red-orange light (prestimulus). We observed that pulses given with a suitable delay after the prestimulus produced unusual responses. Pulses of blue, green, or red-orange light, each eliciting no response when applied alone, produced a secondary photophobic response when applied several seconds after the prestimulus; the same occurred with a negative blue pulse (rapid shut-off and turning on of a blue light). Conversely, no secondary photophobic response was observed when the test stimulus was a step (a step-up for red-orange light, a step-down for blue light) of the same wavelength and intensity. When the delay was varied, different results were obtained with different wavelengths; red-orange pulses were typically effective in producing a secondary photophobic response, even with a delay of 2 s, whereas the response to a blue pulse was suppressed when the test stimulus was applied within 5 s after the prestimulus. The secondary photophobic response to pulses was abolished by reducing the intensity of the prestimulus without affecting the primary photophobic response. These results, some of which were previously reported in the literature as inverse effects, must be produced by a facilitating mechanism depending on the prestimulus itself, the occurrence of reversals being per se ineffective. The fact that red-orange test stimuli are facilitated even at the shortest delay, whereas those of different wavelengths become effective only after several seconds, suggests that the putative mechanism of the facilitating effect is specific for different signaling pathways.

Photoreceptor sensitivity and the shot noise of chemical processes.

The general modeling of dose-response curves to very low stimuli in a photosensory-effector system is critically reshaped starting from basic assumptions on the fluctuations of chemical signals inside the receptor cell, which add to those of the stimulus itself, both arising from their granular (or quantal) structure. We have shown, both through the analytical treatment of a simple kinetic scheme and by means of Monte Carlo simulations of the same, that shot noise arising from chemical transduction ("chemical shot noise") contributes considerably to the output noise of the receptor-effector system, thus affecting both the shape and the abscissa shift of dose-response curves under these conditions; the latter phenomenon has indeed been reported in Halobacterium halobium. After evaluating the general properties of a single-step amplifying mechanism, the effects of introducing several low-amplifying steps in cascade were investigated briefly. The results obtained were qualitatively and quantitatively at variance from those of earlier models on the same phenomenon, and the discrepancies are discussed in order to highlight the fundamental contribution of chemical shot noise to the response of any kind of sensory system to very low stimuli.

Periodic forcing of a K+ channel at various temperatures.

The random sequence of openings and closings of single ion channels and the channel conductances have been the object of intense study over the past two decades with a view toward illuminating the underlying kinetics of the channel protein molecules. Channels that are sensitive to voltage, such as many K(+)-selective channels, have been particularly useful, because the kinetic rates can be manipulated by changing the membrane voltage. Most such studies have been performed under stationary conditions and usually at a single temperature. Here we report the results of experiments with sinusoidal modulation of the membrane potential performed at several temperatures. Dwell time and cycle histograms, objects not normally associated with ion channel experiments, are herein reported. From the last, the transition probability densities for channel opening and closing events are obtained. A new and unusual phase anticipation is observed in the cycle histograms, and its temperature dependence is measured.

Photobehavior of Halobacterium halobium: sinusoidal stimulation and a suppression effect of responses to flashes.

Sequences of reversals recorded by single-cell observation of Halobacterium halobium are analyzed. Autocorrelation functions of spontaneous and stimulated reversals are computed; the results show that the only periodicity present in our data is that of the stimulus. Several different patterns of light stimuli were used. Responses to repetitive linear ramps of different slopes and to sinusoidal lights with different mean values and/or modulation depths are reported, showing that the modulation depth is the stimulus parameter most effective in eliciting photoresponses. Responses to more complex stimuli obtained by superimposing flashes to sinusoidal stimuli are also reported; a suppression effect depending on the phase of the sinusoidal stimulation is shown in responses to complex stimuli. A model which accounts for this effect is proposed.

Use of conditioned distributions in the analysis of ion channel recordings.

A method to test the Markov nature of ion channel gating is proposed. It makes use of singly and doubly conditional distributions. The application of this method to recordings from single BK channels provides evidence that at least two states of the underlying kinetic scheme are left at a constant rate. Moreover, the probabilities, when leaving a state, of reaching another given state are shown to be constant for all the states of the system.

Motor responses of Halobacterium halobrium to sinusodial light stimuli.

Sinusoidal light stimuli have been used to elicit photoresponses in Halobacterium halobium at various periods of the stimulus. Cycle histograms and histograms of the times between consecutive reversals are used to illustrate the responses, and the method used to obtain sinusoidally modulated intense light (with periods up to 1 s) is described.

Effects of current pulses on the sustained discharge of visual cells of Limulus.

Current pulses were used in the eccentric and retinular cells of the Limulus lateral eye to produce changes in the interspike interval of the discharge sustained by a constant light level. The effects on the interspike interval of hyperpolarizing and depolarizing perturbations, applied at various delays from the previous spike, were measured for different intensities and durations of the current pulse. The results show that when the perturbations were applied in the first part of the interval, effects contrary to what is normal were produced (i.e, hyperpolarizing pulses decreased the interspike interval instead of increasing it and vice versa for depolarizing pulses). Here we discuss briefly the implications on neural encoding models.

Resonant response of a neural model and of Limulus ommatidia to double frequency stimulation.

Signals which are the sum of two sinusoids of frequencies v1 and v2 are used to stimulate: i) an electronic analog of the leaky integrator neural model, ii) the visual neurons of the Limulus lateral eye. This makes it possible to investigate the resonant amplification of the impulse density modulation for v1 + v2 which approaches the free-run discharge rate; this resonance is predicted by the Volterra series representation of the leaky integrator (Poggio and Torre, 1977). The resonant responses obtained look very similar for the simulated discharge and for the experimental one.

Effects of electromagnetic fields on the motion of Euglena gracilis.

The orientation behavior of Euglena gracilis cultures in electromagnetic fields is shown to agree with the predictions of a model involving only a passive mechanism. The increase in Euglena motor activity with increasing field intensity is demonstrated by measuring various motion parameters by the laser scattering technique. The effect of electric field on the speed of Euglenas is compared with that of temperature. We conclude that the electric field warms up the culture, thus inducing an increase in cell motility.

Phase-locked responses in the Limulus lateral eye. Theoretical and experimental investigation.

The 1:1 phase locking of the neural discharge to sinusoidally modulated stimuli was investigated both theoretically and experimentally. On the theoretical side, a neural encoder model, the self-inhibited leaky integrator, was considered, and the phase of the locked impulse was computed for each frequency in the locking range by imposing the condition that the "leaky integral" u(t) of the driving signal should reach the threshold for the first time one stimulus period after the preceding impulse. As u(t) can be a nonmonotonic function, this approach leads to results that sometimes differ from those reported in the literature. It turns out that the phase excursion is often much smaller than the values of about 180 degrees predicted from previous analysis. Moreover, our analysis shows a peculiar effect; the phase locking frequency range narrows when the input modulation depth increases. The theoretical predictions are then compared with phase-locked discharge patterns recorded from visual cells of the Limulus lateral eye, stimulated by sinusoidally modulated light or depolarizing current. The phases of the locked spikes at each of a number of modulation frequencies have been measured. The predictions offered by the model fit the experimental data, although there are some difficulties in determining the effective driving signal.