Linear Relations between Stimulus Amplitudes and Amplitudes of Retinal Action Potentials from the Eye of the Wolf Spider

Incremental photic stimuli have been used to elicit small amplitude retinal action potentials from light-adapted ocelli of the wolf spider, Lycosa baltimoriana (Keyserling) in order to see whether or not the amplitudes of these potentials are linearly related to the stimulus amplitudes. Sine wave variations of light intensity around a mean elicit sine wave variations in potential which contain inappreciable harmonics of the stimulus frequency and whose amplitudes are linearly related to the stimulus amplitudes. Likewise, the responses to the first two periodic Fourier components of incremental rectangular wave stimuli of variable duty cycle are directly proportional to the amplitudes of these components and have phases dependent only on the frequencies and phases of these components. Thirdly, a linear transfer function can be found which describes the amplitudes and phases of responses recorded at different frequencies of sine wave stimulation and this transfer function is sufficient to predict the responses to incremental step stimuli. Finally, it is shown that flash response amplitudes are linearly related to incremental flash intensities at all levels of adaptation. The relations of these linear responses to non-linear responses and to physiological mechanisms of the eye are discussed.     

Detection of red and green flashes: evidence for cancellation and facilitation.

Green and red flashes of light will differentially stimulate the middle- and long-wavelength sensitive cones. Interaction of cone signals was studied by measuring increment thresholds for combinations of green and red flashes on a yellow adapting field. When the yellow adapting field was at 10.000 trolands (td), green and red incremental flashes (1 degree, 200-msec duration) produced cancellation when presented simultaneously and facilitation when presented sequentially. A green incremental flash (1.15 degrees, 200 msec, 5000-td adaptation field) and red decremental flash, or vice versa, produced facilitation when presented simultaneously. The results can be explained by color-differencing, opponent-mechanisms. The cancellation effect for the simultaneous incremental flashes largely disappeared when the flashes were exposed briefly (10 msec) or reduced in size (0.04 degrees). It is unlikely that the stimuli were exclusively detected by achromatic, luminance channels, as suggested by previous work, since observers could partially distinguish the hue of threshold flashes of 570- and 590-nm light (0.04 degrees, 10 msec) on a bright yellow field.     

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.     

Light-induced changes of sensitivity in Limulus ventral photoreceptors

The responses of Limulus ventral photoreceptors to brief test flashes and to longer adapting lights were measured under voltage clamp conditions. When the cell was dark adapted, there was a range of energy of the test flashes over which the peak amplitude of the responses (light-induced currents) was directly proportional to the flash energy. This was also true when test flashes were superposed on adapting stimuli but the proportionality constant (termed peak currently/photon) was reduced. The peak current/photon was attenuated more by brighter adapting stimuli than by less bright adapting stimuli. The peak current/photon is a measure of the sensitivity of the conductance-increase mechanism underlying the light response of the photo-receptor. The response elicited by an adapting stimulus had a large initial transient which declined to a smaller plateau. The peak current/photon decreased sharply during the declining phase of the transient and was relatively stable during the plateau. This indicates that the onset of light adaptation is delayed with respect to the onset of the response to the adapting stimulus. If the adaptational state just before the onset of each of a series of adapting stimuli was constant, the amplitude of the transient was a nearly linear function of intensity. When the total intensity was rapidly doubled (or halved) during a plateau response, the total current approximately doubled (or halved). We argue that the transition from transient to plateau, light-elicited changes of threshold, and the nonlinear function relating the plateau response to stimulus intensity all reflect changes of the responsiveness of the conductance-increase mechanism.     

Enhancement and phototransduction in the ventral eye of limulus

Limulus ventral photoreceptors were voltage clamped to the resting (dark) potential and stimulated by a 20-ms test flash and a 1-s conditioning flash. At a constant level of adaptation, we measured the response to the test flash given in the dark (control) and the incremental response produced when the test flash occurred within the duration of the conditioning flash. The incremental response is defined as the response to the conditioning and test flashes minus the response to the conditioning flash given alone. When the test flash was presented within 100 ms after the onset of the conditioning flash we observed that: (a) for dim conditioning flashes the incremental response equaled the control response; (b) for intermediate intensity conditioning flashes the incremental response was greater than the control response (we refer to this as enhancement); (c) for high intensity conditioning flashes the incremental response nearly equaled the control response. Using 10-μm diam spots of illumnination, we stimulated two spatially separate regions of one photoreceptor. When the test flash and the conditioning flash were presented to the same region, enhancement was present; but when the flashes were applied to separate regions, enhancement was nearly absent. This result indicates that enhancement is localized to the region of illumination. We discuss mechanisms that may account for enhancement.     

Dynamics of skate horizontal cells

The all-rod retina of the skate (Raja erinacea or R. oscellata) is known to have the remarkable capability of responding to incremental flashes superimposed on background intensities that initially block all light-evoked responses and are well above the level at which rods saturate in mixed rod/cone retinas. To examine further the unusual properties of the skate visual system, we have analyzed responses of their horizontal cells to intensity-modulated step, sinusoidal, and white-noise stimuli. We found that during exposures to mean intensities bright enough to block responses to incremental stimuli, decremental stimuli were also initially blocked. Thereafter, the horizontal cells underwent a slow recovery phase during which there was marked nonlinearity in their response properties. The cell first (within 2-3 min) responded to decrements in intensity and later (after greater than 10 min) became responsive to incremental stimuli. After adaptation to a steady state, however, the responses to intensity modulation were nearly linear over a broad range of modulation depths even at the brightest mean levels of illumination. Indeed, examination of the steady-state responses over a 5-log-unit range of mean intensities revealed that the amplitude of the white noise-evoked responses depended solely on contrast, and was independent of the retinal irradiance as the latter was increased from 0.02 to 20 muW/cm2; i.e., contrast sensitivity remained unchanged over this 1,000-fold increase in mean irradiance. A decrement from the mean as brief as 2 s, however, disturbed the steady state. Another unexpected finding in this all-rod retina concerns surround-enhancement, a phenomenon observed previously for cone-mediated responses of horizontal cells in the retinas of turtle and catfish. While exposure to annular illumination induced response compression and a pronounced sensitivity loss in response to incremental light flashes delivered to the dark central region, the cell's sensitivity showed a significant increase when tested with a white noise or sinusoidally modulated central spot. Unlike horizontal cells in other retinas studied thus far, however, response dynamics remained unchanged. Responses evoked either by a small spot (0.25-mm diam) or by a large field light covering the entire retina were almost identical in time course. This is in contrast with past findings from cone-driven horizontal cells whose response waveform (dynamics) was dependent upon the size of the retinal area stimulated.     

The Rat Electroretinogram : II. Bloch's law and the latency mechanism of the b-wave

Electroretinograms were obtained from the all-rod eye of the rat with uniform illumination of the entire retina and stimulus flashes of less than 3 msec. duration. Bloch's law of temporal summation was verified for the b-wave latency by varying the time between two equal intensity flashes and observing that no change occurred in the latency when measured from the midpoint of the two flashes. The results of this and other experiments are described in terms of a simple but general model of the latency-determining mechanism. It is shown that this latency mechanism acts as if it depends on a linear additive process; and also that a hypothetical excitatory substance which triggers activity in the sources of the b-wave must accumulate rapidly in time after the flash, approximately as t⁸. The rate at which this substance accumulates is accurately represented by the diffusion equation for more than 4 to 6 log units in the flash intensity. This suggests that the rate-determining step in the latency mechanism may be diffusion-limited.     

The statistical detection of threshold signals in the retina

1. Photographic records of impulses from single ganglion cells in the cat's retina were made while the retina was stimulated by flashes occurring once a second. Ten flashes at each of several intensities near threshold were used. 2. For the purpose of statistical analysis, the number of impulses (x) falling within a critical period following each flash was used as an index of the response. Histograms of x were plotted and used to calculate rates of transfer of information by the ganglion cell for the case of an ideal experiment, the yes-no choice, in which flashes of intensity I and blanks are to be distinguished. 3. The information rate increased (a) with increasing stimulus intensity and (b) with the number of identical flashes or blanks presented successively in a block. The intensity chosen as threshold by the experimenter, who observed the impulses visually and aurally, corresponded to an average information rate for single flashes of 0.7 bit/flash, compared to the maximum possible rate of 1 bit/flash. A threshold intensity giving 0.4 or more bit/flash, if presented in blocks of six identical flashes, corresponded to 0.95 or more bit/block, or near certainty. Thus the calculation of information rates using the index x provides an estimate of threshold at least as sensitive as those obtained during an experiment, which were made only after observing the responses to five to ten flashes of the same intensity. 4. The index x has statistical properties similar to those of the "index of neural activity" used by Tanner and Swets (1954) in their statistical model of human vision, and represents a possible physical interpretation of their index. However, x gave values (0.5 to 1.5) of the parameter called the slope which were consistently smaller than their values (2.1 to 3.1).     

Response entrainment of movement fibers in the optic tract of crayfish

The response properties of jittery movement fibers (JMF) in the crayfish optic tract reacting to a non-moving temporally patterned light were analyzed. The JMFs usually show no response during the regular flickering of stationary light with a flash duration of less than 50 msec when the stimulus frequency is between 4 and 20 per second; however they do respond when the flickering stops if a certain number of flashes have been given. The response appears about 50 msec after the first missing flash, i.e., the latency of the response after the last flash of the train changed from 100 to 300 msec. Thus, the “off” response at the end of the flicker is entrained to the stimulus repetition interval and locked onto the time of the first missing flash. The response of a sustaining fiber to an identical stimulus has quite different features as illustrated in Fig. 2. Some of the fibers show responses to the beginning part of the flicker but not necessarily to each flash, and habituate after several flashes. When a single flash longer than 250 msec is given, the fiber shows an “off” response with about 50 msec latency, as it does to sustained light. Some fibers show a double burst of “off” discharge to single flashes; the first at 50 msec is followed after 120 msec by the second one. However, when the flash duration is between 250 and 50 msec, a single flash elicits little or no response. The latency of the “off” response is as much as 300 msec for short single flashes less than 50 msec. An “on” response to flashes of light is observed when the inter-stimulus interval is more than 5 sec. The responses to the beginning part of flicker train are not simply locked to the just preceding flash except the “on” response to the very first one, but they can be the long latency responses to the flash before that. This response is modified in latency by the succeeding flashes in flicker trains and becomes entrained to the missing flash. Four types of entrainment are classified on the basis of the change in latency from the missing flash with regard to the number of flashes in a train. In most cases, 10 flashes are sufficient to entrain the response to the first missing flash. Non-resposiveness, i.e., habituation, during a regular flicker, may be due to an active inhibitory process, initiated by each succeeding light pulse. The response to the missing flash, therefore results from a disinhibited modified response to the last flash. Some JMFs continue to respond to the flicker even after a considerable number of flashes but only when the repetition interval is about 120 msec corresponding well to the interval of the double burst “off” discharge, thus the JMF has a resonant frequency of about 8 Hz. The JMFs appear to be acting as an irregularity detector in temporal sequence.     

Abdomen Posture and Flash Gesture Direction in a Female Synchronic Firefly <Emphasis Type="Italic">Photinus carolinus</Emphasis> (Green) (Coleoptera: Lampyridae)

North American Photinus fireflies use bioluminescent flashes to communicate an individual’s species and sex, and to attract potential mates. A female firefly responds to a male firefly’s courtship flash with her own species-specific flash. We used a photic stimulator to produce male-like species-specific P. carolinus LED courtship flashes. These evoked species-specific response flashes from a female. The female’s flashes were preceded by a flash gesture comprising a sequence of abdominal postural adjustments (pitch, roll, and yaw). These gestures changed her lantern’s orientation which, at rest, was downward towards the substrate. Our results demonstrate that these gestures mediate a lateralization of the female’s response flashes towards the direction of the stimulating LED. That is, she directs her response to the left of midline when stimuli are presented from her left, and similarly, she directs her response to the right of midline when stimuli are presented from her right. The directional aspect of the flash gesture adds a new perspective to the complexity of the behaviors associated with flash communication in fireflies. Lateralization of the flash gesture suggests that the female’s visual system processes information about the location of male’s flashes as well as their temporal pattern.     

Mechanisms and asymmetries in visual perception of simultaneity and temporal order

Temporally overlapping, spatially separated visual stimuli were used for studying perception of simultaneity and temporal order. Pairs of flashes each of 100 ms duration were presented with stimulus onset asynchronies of 0, 30, 50, and 70 ms. Three spatial arrangements of flash presentation were tested: 1) both flashes were presented foveally; 2) one flash was presented foveally and the other at 9 deg in the left visual hemifield; 3) one flash was presented foveally and the other at 8 deg in the right visual hemifield. Onset asynchronies of 30 and 50 ms were not sufficient for correct identification of temporal order although the flashes were not perceived as simultaneous. Analysis of the response distributions suggests the existence of two-independent mechanisms for evaluating temporal interrelations: one for detecting simultaneity and the other for identifying temporal order. A better detection of simultaneity was found when synchroneous flashes were presented together with pairs of flashes separated by larger onset asynchronies. Reading habits may explain only part of the left-right asymmetries of the response distributions. The possible lateralization of the two suggested mechanisms within the cerebral hemispheres is discussed.     

Mechanisms of synchrony in the North American fireflyPhotinus carolinus (Coleoptera: Lampyridae)

Photometric recordings combined with computer stimulation, acquisition and analysis were used to study synchrony in the North American fireflyPhotinus carolinus. A computer-generated burst of simulatedP. carolinus flashes was used to trigger a firefly flash burst. We found that the first triggered firefly flashes occurred after the second or third flashes in the stimulus burst, that there were fewer flashes in a triggered burst than a spontaneous burst, and that extending the stimulus flashes into the firefly's interburst interval inhibited firefly flashing. When the stimulus flash interval (389–560 ms) was changed, no change was seen in the interflash interval. When the stimulus flash interval was changed, the average time between stimulus flash and firefly flash (flash delay) changed as if the firefly interflash interval was constant. Thus, interflash interval inP. carolinus does not change its length, making it similar to the Southeast Asian synchronizerPteroptyx cribellata and different fromPteroptyx malaccae, which can change its interval. We suspect that the time between bursts is functionally analogous to the time between flashes in Southeast Asian synchronizers.     

The Impact of Spatial Incongruence on an Auditory-Visual Illusion

Background

The sound-induced flash illusion is an auditory-visual illusion – when a single flash is presented along with two or more beeps, observers report seeing two or more flashes. Previous research has shown that the illusion gradually disappears as the temporal delay between auditory and visual stimuli increases, suggesting that the illusion is consistent with existing temporal rules of neural activation in the superior colliculus to multisensory stimuli. However little is known about the effect of spatial incongruence, and whether the illusion follows the corresponding spatial rule. If the illusion occurs less strongly when auditory and visual stimuli are separated, then integrative processes supporting the illusion must be strongly dependant on spatial congruence. In this case, the illusion would be consistent with both the spatial and temporal rules describing response properties of multisensory neurons in the superior colliculus.

Methodology/Principal Findings

The main aim of this study was to investigate the importance of spatial congruence in the flash-beep illusion. Selected combinations of one to four short flashes and zero to four short 3.5 KHz tones were presented. Observers were asked to count the number of flashes they saw. After replication of the basic illusion using centrally-presented stimuli, the auditory and visual components of the illusion stimuli were presented either both 10 degrees to the left or right of fixation (spatially congruent) or on opposite (spatially incongruent) sides, for a total separation of 20 degrees.

Conclusions/Significance

The sound-induced flash fission illusion was successfully replicated. However, when the sources of the auditory and visual stimuli were spatially separated, perception of the illusion was unaffected, suggesting that the “spatial rule” does not extend to describing behavioural responses in this illusion. We also find no evidence for an associated “fusion” illusion reportedly occurring when multiple flashes are accompanied by a single beep.     

Fast visuomotor processing of redundant targets: the role of the right temporo-parietal junction

Parallel processing of multiple sensory stimuli is critical for efficient, successful interaction with the environment. An experimental approach to studying parallel processing in sensorimotor integration is to examine reaction times to multiple copies of the same stimulus. Reaction times to bilateral copies of light flashes are faster than to single, unilateral light flashes. These faster responses may be due to 'statistical facilitation' between independent processing streams engaged by the two copies of the light flash. On some trials, however, reaction times are faster than predicted by statistical facilitation. This indicates that a neural 'coactivation' of the two processing streams must have occurred. Here we use fMRI to investigate the neural locus of this coactivation. Subjects responded manually to the detection of unilateral light flashes presented to the left or right visual hemifield, and to the detection of bilateral light flashes. We compared the bilateral trials where subjects' reaction times exceeded the limit predicted by statistical facilitation to bilateral trials that did not exceed the limit. Activity in the right temporo-parietal junction was higher in those bilateral trials that showed coactivation than in those that did not. These results suggest the neural coactivation observed in visuomotor integration occurs at a cognitive rather than sensory or motor stage of processing.     

Neural Responses in Parietal and Occipital Areas in Response to Visual Events Are Modulated by Prior Multisensory Stimuli

The effect of multi-modal vs uni-modal prior stimuli on the subsequent processing of a simple flash stimulus was studied in the context of the audio-visual ‘flash-beep’ illusion, in which the number of flashes a person sees is influenced by accompanying beep stimuli. EEG recordings were made while combinations of simple visual and audio-visual stimuli were presented. The experiments found that the electric field strength related to a flash stimulus was stronger when it was preceded by a multi-modal flash/beep stimulus, compared to when it was preceded by another uni-modal flash stimulus. This difference was found to be significant in two distinct timeframes – an early timeframe, from 130–160 ms, and a late timeframe, from 300–320 ms. Source localisation analysis found that the increased activity in the early interval was localised to an area centred on the inferior and superior parietal lobes, whereas the later increase was associated with stronger activity in an area centred on primary and secondary visual cortex, in the occipital lobe. The results suggest that processing of a visual stimulus can be affected by the presence of an immediately prior multisensory event. Relatively long-lasting interactions generated by the initial auditory and visual stimuli altered the processing of a subsequent visual stimulus.     

On the interactivity in the human visual cortex caused by specific and nonspecific inflows

Summary An interactivity (Sato, 1969) with respect not only to the interstimulus intervenal between the conditioned and test stimuli but also with respect to the time after the test stimulus was defined as a new generalized recovery function.The contour maps of the above interactivities of the cerebral evoked potentials elicited by the paired flash stimuli and by the conditioned cutaneous and test flash stimuli, respectively, were obtained in the adult human visual cortex.It was suggested to a certain degree by the autocorrelograms of background occipital EEGs and average VEPs that the more prominent VEPs are elicited, the more predominant are the alpha waves in the occipital background EEG, and vice versa. In the contour maps of the interactivity by the paired flash stimuli, cyclic changes of facilitory mountains and occlusive valleys were regular and conspicuous in the subject who exhibited predominant alpha waves in his background EEG, whereas the cyclic changes were less regular and less marked in the subject with few alpha waves in the background EEG.The cyclic changes in the former subject appeared at an interval of about 100 msec at an interstimulus interval of around 100 msec. This acts favorably in the contribution of the cyclic change in interactivity via natural stimulations to the generation of alpha waves in spontaneous EEG. The main facilitory mountains and occlusive valleys in the maps were arranged in a line parallel to the ordinate (axis of the interstimulus interval), which suggest that the conditioned flash will affect in a specific fashion each response compoment elicited via specific afferent inflows in the VEP timelocked to the test flash stimulus. On the contrary, those in the maps obtained by the conditioned cutaneous and test flash stimuli were diagonal to the abscissa (time after the test stimulus) and ordinate, and/or the peaks of facilitory mountains and bottoms of occlusive valleys tended to form a line parallel to the abscissa, which indicates that as the cutaneous somatosensory afferent inflow converges to the visual cortex nonspecific interactivities are exhibited in each component in the VEP.     

Normal and mutant rhodopsin activation measured with the early receptor current in a unicellular expression system.

The early receptor current (ERC) represents molecular charge movement during rhodopsin conformational dynamics. To determine whether this time-resolved assay can probe various aspects of structure-function relationships in rhodopsin, we first measured properties of expressed normal human rhodopsin with ERC recordings. These studies were conducted in single fused giant cells containing on the order of a picogram of regenerated pigment. The action spectrum of the ERC of normal human opsin regenerated with 11-cis-retinal was fit by the human rhodopsin absorbance spectrum. Successive flashes extinguished ERC signals consistent with bleaching of a rhodopsin photopigment with a normal range of photosensitivity. ERC signals followed the univariance principle since millisecond-order relaxation kinetics were independent of the wavelength of the flash stimulus. After signal extinction, dark adaptation without added 11-cis-retinal resulted in spontaneous pigment regeneration from an intracellular store of chromophore remaining from earlier loading. After the ERC was extinguished, 350-nm flashes overlapping metarhodopsin-II absorption promoted immediate recovery of ERC charge motions identified by subsequent 500-nm flashes. Small inverted R(2) signals were seen in response to some 350-nm flashes. These results indicate that the ERC can be photoregenerated from the metarhodopsin-II state. Regeneration with 9-cis-retinal permits recording of ERC signals consistent with flash activation of isorhodopsin. We initiated structure-function studies by measuring ERC signals in cells expressing the D83N and E134Q mutant human rhodopsin pigments. D83N ERCs were simplified in comparison with normal rhodopsin, while E134Q ERCs had only the early phase of charge motion. This study demonstrates that properties of normal rhodopsin can be accurately measured with the ERC assay and that a structure-function investigation of rapid activation processes in analogue and mutant visual pigments is feasible in a live unicellular environment.     

Photoreceptor Spike Responses in the Hardshell Clam, Mercenaria mercenaria

Spikes were recorded from single axons in the siphonal nerve of the hardshell clam Mercenaria mercenaria which respond to dimming of light. No axons were found to respond to the onset, or increase, of illumination. In a dark-adapted state there is little or no ongoing spike activity. The responsive area of a single axon is a circle of approximately 85 µm diameter on the inner siphon wall. The number of spikes elicited at the off of constant-duration flashes grows as approximately the 0.4 power of flash intensity. For constant intensity and constant light-time fraction, the off-response increases with increasing duration at least up to 500 s duration. For long durations, the response grows as the logarithm of stimulus duration. Subthreshold light can suppress the off-response from preceding illumination. In a light-adapted state, the off-response is greater and its latency shorter than in the dark-adapted state. The fine structure of groups of cell processes thought to comprise the photoreceptor in Mercenaria is described. On the basis of morphological and physiological findings it is suggested that phototransduction occurs in the fine distal processes of the axons from which we have recorded. Axonal processes were found to contain well organized pentalaminar whorls which may be the site of photo-pigment concentration. The action spectrum obtained from the integrated responses of nerve bundles appears to be that of a single Dartnall pigment having maximal absorption at about 510 nm.     

The induction kinetics of bacterial photophosphorylation. Threshold effects by the phosphate potential and correlation with the amplitude of the carotenoid absorption band shift

1. ATP synthesis (monitored by luciferin-luciferase) can be elicited by a single turnover flash of saturating intensity in chromatophores from Rhodopseudomonas capsulata, Kb1. The ATP yield from the first to the fourth turnover is strongly influenced by the phosphate potential: at high phosphate potential (?11.5 kcal/mol) no ATP is formed in the first three turnovers while at lower phosphate potential (?8.2 kcal/mol) the yield in the first flash is already one half of the maximum, which is reached after 2–3 turnovers.2. The response to ionophores indicates that the driving force for ATP synthesis in the first 20 turnovers is mainly given by a membrane potential. The amplitude of the carotenoid band shift shows that during a train of flashes an increasing ΔΨ is built up, which reaches a stationary level after a few turnovers; at high phosphate potential, therefore, more turnovers of the same photosynthetic unit are required to overcome an energetic threshold.3. After several (six to seven) flashes the ATP yield becomes constant, independently from the phosphate potential; the yield varies, however, as a function of dark time (t_d) between flashes, with an optimum for t_d = 160–320 ms.4. The decay kinetics of the high energy state generated by a long (125 ms) flash have been studied directly measuring the ATP yield produced in post-illumination by one single turnover flash, under conditions of phosphate potential (?10 kcal/mol), which will not allow ATP formation by one single turnover. The high energy state decays within 20 s after the illumination. The decay rate is strongly accelerated by 10^?8 M valinomycin.5. Under all the experimental conditions described, the amplitude of the carotenoid signal correlates univocally with the ATP yield per flash, demonstrating that this signal monitores accurately an energetic state of the membrane directly involved in ATP synthesis.6. Although values of the carotenoid signal much larger than the minimal threshold are present, relax slowly, and contribute to the energy input for phosphorylation, no ATP is formed unless electron flow is induced by a single turnover flash.7. The conclusions drawn are independent from the assumption that a ΔΨ between bulk phases is evaluable from the carotenoid signal.     

The initial response of Limulus ventral photoreceptors to bright flashes. Released calcium as a synergist to excitation

The leading edge of the response of Limulus ventral photoreceptors to brief flashes was investigated using a voltage clamp. The leading edge of responses increases linearly with flash intensity when dim flashes produce less than one photoisomerization per square micron of cell surface. Brighter flashes accelerate the initial portion of the response, resulting in a fourth-power relationship between the magnitude of the response at brief times after the flash and the flash intensity. The onset of this nonlinearity with increasing flash intensity is determined by the local density of photoisomerizations within the receptor. Responses to bright 10-15-mum-diam spots therefore rise faster than responses to diffuse flashes producing the same number of photoisomerizations within the receptor. Background illumination shortens the response latency and suppresses the initial nonlinearity. These phenomena can be explained by a model of transduction in which light activates two parallel cascades of reactions. Particles released by the first of these cascades open ionic channels, while the second produces an agent that accelerates the rate of production of particles by the first. Injection of the calcium buffer EGTA slows the initial portion of the response to bright flashes and suppresses its nonlinearity, which suggests that the accelerating agent released by the second cascade is calcium.