PHOTOTACTIC BEHAVIOR OF INDIVIDUAL CELLS OF CRYPTOMONAS SP. IN RESPONSE TO CONTINUOUS AND INTERMITTENT LIGHT STIMULI

Abstract— The phototactic response of cells of Cryptomonas sp. to stimulation with continuous or intermittent lateral light was determined by an individual cell method using photomicrography and videomicrography. The cells showed positive phototaxis under the conditions studied. The phototactic orientation of individual cells was induced most effectively by irradiation with light of 570 nm; blue light was less effective, and no orientation was found in red light. An intermittent stimulus regime with a long dark interval (250 ms) elicited a weaker phototactic orientation than did a regime with a short dark interval (63 ms) irrespective of the duration of light pulses (16, 250 and 1000 ms). The swimming rate was ca. 240 ums ^-1 and the rotation period ca. 450 ms in the dark, neither of which was greatly affected by stimulation with continuous or intermittent light. Neither step-up nor step-down photophobic responses were observed at the time of onset or removal of the light stimulus under the experimental conditions. The swimming direction of individual cells became gradually oriented toward the light source. Phototactic response was detectable within 4 s after the onset of light stimulation, reaching a saturation level after more than 30 s.     

Dark adaptation of horizontal cells in the teleost fish retina

The dark adaptation behaviors of rod-driven and cone-driven horizontal cells were examined by analyzing their light responses recorded intracellularly in the intact, immobilized carp, and compared with that of the electroretinographic b-wave recorded simultaneously. Like the b-wave, the light responsiveness of rod horizontal cells increased gradually with time in the dark and attained a steady level at 60 min. On the other hand, cone horizontal cells initially increased in light responsiveness in the first 10 min, but thereafter decreased steadily so that the response amplitudes of these cells to bright light flashes were only 3-5 mV. The results suggest that cone horizontal cells are strongly suppressed in prolonged darkness.     

LATERAL INTERACTIONS BETWEEN VERTEBRATE PHOTORECEPTORS

Vertebrate photoreceptors generate electrical signals across their cell membrane when they absorb light. Recent studies show, moreover, that the membrane potential of an individual photoreceptor may be also modified by illumination of its neighbors. Two types of lateral interactions have been described: a direct interaction mediated by electrical synapses between adjacent photoreceptors, and a recurrent interaction mediated by a feedback circuit involving horizontal cells. In the first mechanism, photoreceptors summate their responses over short retinal distances. In the feedback circuit photoreceptors may develop light responses of opposite polarity to those induced by direct illumination. In general, these 'antagonistic' responses are best elicited by large area retinal illumination. As a consequence of these photoreceptor interactions rather complex processing of visual information, in both spatial and chromatic domains, occurs at the first stage of the retinal network.     

DARK ADAPTATION OF HORIZONTAL CELLS IN THE TELEOST FISH RETINA

The dark adaptation behaviors of rod-driven and cone-driven horizontal cells were exam-ined by analyzing their light responses recorded intracellularly in the intact, immobilizedcarp, and compared with that of the electroretinographic b--wave recorded simultaneously.Like the b--wave, the light responsiveness of rod horizontal cells increased gradually withtime in the dark and attained a steady level at 60 min. On the other hand, cone horizontalcells initially increased in light responsiveness in the first 10 min, but thereafter decreasedsteadily so that the response amplitudes of these cells to bright light flashes were only 3--5 mV.The results suggest that cone horizontal cells are strongly suppressed in prolonged darkness.     

THE VECTORIAL PHOTO-EXCITATION IN SOLAR-TRACKING LEAVES OF Lavatera cretica (MALVACEAE)*,†

Abstract— The diaphototropic responses of the solar-tracking leaves of Lavatera cretica were studied under constant levels of vectorial photo-excitation (negligible variations in angle of incidence and fluence rates). The results showed the following:
(a) The photoreceptors, that are associated with the (major) veins, can equally perceive vectorial excitation (by an oblique light-beam in the plane of symmetry of the vein), directed either towards its tip (tip-oriented: TO), or towards its base (base-oriented: BO).
(b) The mechanism of photo-perception is apparently qualitatively the same for TO and BO excitation.
(c) The response to TO and BO vectorial excitation, which takes place in a circular sheath of motor-cell tissue in the pulvinus (a2–3 mm long segment at the top of the petiole), is also highly directional and results in bending in the vertical plane of the oblique beam. Each sector of the motor tissue can expand longitudinally in response to TO excitation, presumably by influx of solutes, and contract longitudinally in response to BO excitation of the same vein, presumably by efflux of solutes.
(d) The angular velocity of the vectorial response to a constant level of vectorial excitation remains constant over large angular displacements and long periods, and was similar for TO and BO over a wide range of fluence rates (30-400 μmol m⁻²s⁻¹) of white light (400-700 nm).
(e) Reversal of the direction of vectorial excitation (TO to BO, and vice versa) results in reversal of the direction of the vectorial response (laminar reorientation). The change in the direction of laminar reorientation involved a larger overshoot in the original direction when the change was from BO to TO, than in the reverse case.     

Phototransduction: different mechanisms in vertebrates and invertebrates

The photoreceptor cells of invertebrate animals differ from those of vertebrates in morphology and physiology. Our present knowledge of the different structures and transduction mechanisms of the two animal groups is described. In invertebrates, rhodopsin is converted by light into a meta-rhodopsin which is thermally stable and is usually re-isomerized by light. In contrast, photoisomerization in vertebrates leads to dissociation of the chromophore from opsin, and a metabolic process is necessary to regenerate rhodopsin. The electrical signals of visual excitation have opposite character in vertebrates and invertebrates: the vertebrate photoreceptor cell is hyperpolarized because of a decrease in conductance and invertebrate photoreceptors are depolarized owing to an increase in conductance. Single-photon-evoked excitatory events, which are believed to be a result of concerted action (the opening in invertebrates and the closing in vertebrates) of many light-modulated cation channels, are very different in terms of size and time course of photoreceptors for invertebrates and vertebrates. In invertebrates, the single-photon events (bumps) produced under identical conditions vary greatly in delay (latency), time course and size. The multiphoton response to brighter stimuli is several times as long as a response evoked by a single photon. The single-photon response of vertebrates has a standard size, a standard latency and a standard time course, all three parameters showing relatively small variations. Responses to flashes containing several photons have a shape and time scale that are similar to the single-photon-evoked events, varying only by an amplitude scaling factor, but not in latency and time course. In both vertebrate and invertebrate photoreceptors the single-photon-evoked events become smaller (in size) and faster owing to light adaptation. Calcium is mainly involved in these adaptation phenomena. All light adaptation in vertebrates is primarily, or perhaps exclusively, attributable to calcium feedback. In invertebrates, cyclic AMP (cAMP) is apparently another controller of sensitivity in dark adaptation. The interaction of photoexcited rhodopsin with a G-protein is similar in both vertebrate and invertebrate photoreceptors. However, these G-proteins activate different photoreceptor enzymes (phosphodiesterases): phospholipase C in invertebrates and cGMP phosphodiesterase in vertebrates. In the photoreceptors of vertebrates light leads to a rapid hydrolysis of cGMP which results in closing of cation channels. At present, the identity of the internal terminal messenger in invertebrate photoreceptors is still unsolved.(ABSTRACT TRUNCATED AT 400 WORDS)     

PHYSIOLOGICAL STUDIES ON PEA TENDRILS—X. CHARACTERIZATION OF THE LIGHT ACTIVATION EFFECT ON CONTACT COILING AS A BLUE LIGHT TRIGGER*

Abstract— Tendrils of the Alaska pea lose their capacity to coil in response to a mechanical stimulus between 24 and 48 h after being placed in the dark. Light is thereafter necessary for the motor response to proceed once the mechanical stimulus has becn sensed by dark-adapted tendrils. An action spectrum for this light activation effect showed that the most effective region of the visible spectrum was a narrow band in the blue region. Tendrils excised from their petiole and leaflets coiled more than any other preparation used. A linear relationship was found between the duration of post-stimulated white light treatment and the amount of coiling.     

Evidence for a circadian rhythm of susceptibility to retinal light damage

This study investigated a possible circadian rhythm of light damage susceptibility in photoreceptors of both cyclic light-reared and dark-reared rats. A single exposure to intense green light was administered, beginning either in the early light period, the late light period or the dark period. In some animals exposed in the dark period, the synthetic antioxidant dimethylthiourea was administered before or after the onset of intense light exposure. Retinas were examined either immediately after exposure or after 2 weeks of recovery in darkness. Rod outer segment length and outer nuclear layer thickness measurements were used to assess light damage, along with qualitative analysis of swelling and disruption of the outer retinal layers. In all animals, retinal light damage was the most severe when intense light exposure began during the dark period. However, this severe damage was significantly reduced by pretreatment with the antioxidant. In a separate set of unexposed animals, fluctuations in plasma adrenocorticotropic hormone (ACTH) and corticosterone concentrations followed the same time course, regardless of the light regime during rearing. Our data support the notion of a circadian rhythm of light damage susceptibility that peaks in the dark period and yet can be modulated by the exogenous administration of an antioxidant.     

EXTRAOCULAR PHOTORECEPTION IN AMPHIBIANS

Abstract— Amphibians possess extraocular photoreceptors (EOPs) which exclusively or together with the lateral eyes perceive light for various physiological and behavioral activities. Several kinds of EOPs are discussed but emphasis is given to the pineal complex: the dermal frontal organ (or stirnorgan) found only in frogs and toads among amphibians and the intracranial pineal body (or epiphysis cerebri ) found in all Amphibia. Both structures are derived as dorsal evaginations of the diencephalon and have a retina-like fine structure. Both are sensitive to visible and UV light but not to IR, mechanical or chemical stimuli. The frontal organ gives chromatic and achromatic responses but in most species only achromatic ones are recorded from the pineal. Photopigments have been identified for some of these responses.
Pineal EOPs are involved in several activities: (i) neurosecretory activity by the subcommissural organ in the brain; (ii) body-lightening reaction in larval amphibians, involving the light-inhibited release by the pineal of a hormone, presumably melatonin, which contracts melanophores in the skin; (iii) cuing of circadian locomotor rhythms, including phase shifts and synchronization with environmental light cycles; (iv) perception of celestial cues for use in time-compensated compass orientation; and (v) possibly perception of linearly-polarized light.
Non-pineal EOPs have been implicated in (i) tail-darkening reaction of frog larvae where light has a direct effect on melanophores; and (ii) in phototaxis where unknown EOPs in various parts of the body and tail mediate withdrawal from light. Evidence for use of pineal and non-pineal EOPs and location of specific receptors is reviewed drawn from biochemical, ultrastructural, neurophysiological and behavioral studies. The possible use of EOPs for other biological functions and their possible adaptive value are briefly discussed.     

Fluorescence resonance energy transfer between polyphenolic compounds and riboflavin indicates a possible accessory photoreceptor function for some polyphenolic compounds

The photoreceptive extreme tip of the wheat coleoptile exhibits intense green-yellow fluorescence under UV light, suggesting the presence of UV-absorbing materials. Fluorescence spectra of the intact coleoptile tip and tip homogenate showed the presence of the known photoreceptor pigments flavin and carotene, and a preponderance of phenolic compounds. Absorption spectra and fluorescence spectra of various phenolic compounds showed close overlap with the absorption and fluorescence spectra of the wheat coleoptile tip homogenate. Fluorescence spectra of several phenolic compounds showed close overlap with the absorption bands of flavin, carotene and pterine, suggesting possible energy transduction from phenols to these photoreceptors. Excitation of gentisic acid and ferulic acid with 340 nm light in the presence of flavin showed enhancement of flavin fluorescence in a concentration- and viscosity-dependent fashion, indicating fluorescence resonance energy transfer between them and riboflavin. Furthermore, several phenolic compounds tested generated superoxide anion on excitation at 340 nm, suggesting that superoxide-dependent signal cascades could operate in a polyphenol-mediated pathway. Phenolic compounds thus may act as accessory photoreceptors bringing about excitation energy transfer to the reactive photoreceptor molecules, or they may take over the function of the normal photoreceptor in genetic mutations lacking the system, or both processes may occur. The responses of plants to UV-B and UV-A light in mutants may be explained in terms of various phenolics acting as energy transducers in photoreceptor functioning.     

INTERACTION BETWEEN PHYTOCHROME AND THE BLUE LIGHT PHOTORECEPTOR SYSTEM IN Mougeotia*

Abstract— Face-to-profile chloroplast movement in Mougeotia was induced by sequences of strong blue and red short irradiations. This type of response occured only when blue light was applied prior to or simultaneously with red light, and far-red irradiation was necessary after the sequence to cancel the remaining gradient of the far-red absorbing form of phytochrome P_fr. The dependence of the response magnitude on blue and red light sequences was studied for a wide range of light durations and dark intervals. The relationship between the response and the dark interval points to the lack of direct coupling between phytochrome and blue-absorbing “cryptochrome”. It was postulated that a photoproduct having a life-time of2–3 min is formed by the blue-light-mediated reaction. This photoproduct interacts with phytochrome during its transformation or with its final P_fr form.     

Bacterial Photosensory Proteins and Their Role in Plant–pathogen Interactions

Light is an important environmental signal for almost all living organisms. The light perception is achieved by photoreceptor proteins. As can be observed from the great number of bacterial genomes sequenced, plant pathogenic bacteria encode for a large number of photoreceptor proteins. The physiological implications of these photoreceptors are still poorly characterized. However, recent studies revealed the participation of these photosensory proteins in the pathogenic process. Here, we summarize what is known about these proteins and their role during the virulence process, concluding that the light environment modulates the plant–pathogen interaction.     

Controlled Sol–Gel Transitions of a Thermoresponsive Polymer in a Photoswitchable Azobenzene Ionic Liquid as a Molecular Trigger

Producing ionic liquids (ILs) that function as molecular trigger for macroscopic change is a challenging issue. Photoisomerization of an azobenzene IL at the molecular level evokes a macroscopic response (light‐controlled mechanical sol–gel transitions) for ABA triblock copolymer solutions. The A endblocks, poly(2‐phenylethyl methacrylate), show a lower critical solution temperature in the IL mixture containing azobenzene, while the B midblock, poly(methyl methacrylate), is compatible with the mixture. In a concentrated polymer solution, different gelation temperatures were observed in it under dark and UV conditions. Light‐controlled sol–gel transitions were achieved by a photoresponsive solubility change of the A endblocks upon photoisomerization of the azobenzene IL. Therefore, an azobenzene IL as a molecular switch can tune the self‐assembly of a thermoresponsive polymer, leading to macroscopic light‐controlled sol–gel transitions.     

Retinal phosphenes and discrete dark noises in rods: A new biophysical framework

Spontaneous rhodopsin activation produces discrete noises indistinguishable from single-photon responses. However, there is a serious discrepancy between the apparent energy barrier of thermal events compared with that of the photon-driven process. Current estimates of the activation energies of discrete dark noises in vertebrate rod and cone pigments are 40–50 kcal/mol for activation by photon and 20–25 kcal/mol for activation by heat. To reconcile this discrepancy, it was assumed that thermal activation and photon activation of rhodopsin follow different molecular mechanisms. The most convincing hypothesis for a separate low-energy thermal pathway is that the discrete dark noises of rods arise in a small subpopulation of rhodopsins, where the Schiff base linking the chromophore to the protein part has been deprotonated.According to Narici et al.’ experiments (2009, Radiation Measurements), phosphene perception in space travel is due to the ionizing radiation-induced free radicals that generate chemiluminescent photons from lipid peroxidation. These photons are absorbed by the photoreceptors chromophores, which modify the rhodopsin molecules (bleaching) and start the photo-transduction cascade resulting in the perception of phosphenes.Here, we point out that not only retinal phosphenes but also the discrete dark noise of rods can be due to the natural redox related (free radical) bioluminescent photons in the retina. In other words, under regulated conditions, lipid peroxidation is a natural process in cells and also in retinal membranes. Since the natural lipid peroxidation is one of the main sources of bioluminescent photons and the photoreceptors have the highest oxygen demand and polyunsaturated fatty acid (PUFA) concentration, there is a continuous, low level bioluminescent photon emission in the retina without any external photonic stimulation. During photopic or scotopic vision, evanescent bioluminescent photon emission is negligible. In contrast, in dark-adapted retinal cells this evanescent bioluminescent photon emission is not negligible. Therefore, our hypothesis is that the discrete dark noise of rods can be due to these bioluminescent photons.     

STATE I-STATE II TRANSITIONS IN THE THERMOPHILIC BLUE-GREEN ALGA (CYANOBACTERIUM) Synechococcus lividus*

Time courses of state I-state II transitions were measured in the thermophilic blue-green alga (Cyanobacterium), Synechococcus lividus, that was grown at 55°C. The rate of the state I–II transition using light II illumination was the same as that in the dark, and the dark state was identified to be state II. Therefore, light regulation attained by state transitions is produced by the state II–I transition induced by system I light. The redox level of plastoquinone did not affect this dark state II. Arrhenius plots of the state transitions showed a break point around 43°C that corresponded to the phase transition temperature of this alga. Since both the state I–II and II–I transitions were very much temperature-independent, we could keep the alga in either state for a long time at a “low” temperature such as room temperature. Activities of both photosystems I and II in states I and II were also measured. After a state II–I transition, the system II activity increased about 16% and at the same time, svstem I activity decreased about 30%.     

INTERMEDIATE PROCESSES IN PHOTOTRANSDUCTION: A STUDY IN DROSOPHILA MUTANTS

Abstract— A variety of procedures were used to modify the light response of Drosophila photoreceptors in order to find out if these manipulations produce effects that mimic some aspects of light adaptation. The ultimate goal of our approach is to use these experimental manipulations to dissect the different stages of the phototransduction process.
The means which were used to modify the light response were as follows: (a) light adaptation of normal photoreceptors, (b) exposure of normal photoreceptors to various levels of CO₂ (c) light adaptation of the trp Drosophila mutant in which the receptor potential decays to baseline during illumination, (d) exposure of the temperature sensitive norpA ^H52 mutant of Drosophila to elevated temperatures. Intensity-response functions were obtained using intracellular and extracellular recordings. The maximal response amplitude ( V _max) was then plotted as a function of the light intensity () which evoked a response of half maximal amplitude. This plot was used to compare the effects of the procedures described above. Qualitatively all the manipulations had similar effects, they reduced V _max and shifted to higher levels of light intensity, however, quantitatively the various effects were different.
The finding that each experimental manipulation gave a different V _max versus plot suggests that they affect different stages in phototransduction. We suggest that the slope of the V _max versus a function can be interpreted in terms of an ordered cascade of events in phototransduction. This interpretation might give us a tool to distinguish between early and late stages in phototransduction.     

Phytochrome-dependent photomovement responses mediated by phototropin family proteins in cryptogam plants

In this review, we describe the regulation of photomovement responses by phototropin and phytochrome photoreceptors. The blue light receptor phototropin mediates various photomovement responses such as phototropism, chloroplast movement and stomatal opening. In cryptogamic plants including ferns, mosses and green alga, red as well as blue light mediates phototropism and chloroplast movement. The red/far-red light reversibility suggests the involvement of phytochrome in these responses. Thereby, plant growth is presumably promoted by coordinating these photomovements to capture efficiently light for photosynthesis.     

DOSE RESPONSE BEHAVIOUR FOR POLAROTROPISM OF THE CHLORONEMA OF THE FERN DRYOPTERIS FILIX-MAS (L.) SCHOTT

Abstract— The dose response behaviour for polarotropism of the chloronema of the fern Dryopteris filix-mas was studied in detail. By varying the two factors, time and intensity, dose response curves were obtained, which consisted of as many as two maxima and minima or no maxima and minima at all. Furthermore, the effects of red light and dark, given before polarotropic induction, were compared, and the influence of temperature on the dose response behaviour was examined. There are two main results: (1) Under all conditions tested, the dose response behaviour in far-red, red, blue, and near U.V. apparently is the same. (2) In this one system, the chloronema of Dryopteris , all types of dose response curves, which have been described previously for tropic responses mediated by light, could be obtained simply by varying the conditions. The variable and complex dose response behaviour is discussed in connection with other photoresponses in lower and higher plants.     

Fractionated illumination after topical application of 5-aminolevulinic acid on normal skin of hairless mice: the influence of the dark interval

We have previously shown that light fractionation during topical aminolevulinic acid based photodynamic therapy (ALA-PDT) with a dark interval of 2h leads to a significant increase in efficacy in both pre-clinical and clinical PDT. However this fractionated illumination scheme required an extended overall treatment time. Therefore we investigated the relationship between the dark interval and PDT response with the aim of reducing the overall treatment time without reducing the efficacy. Five groups of mice were treated with ALA-PDT using a single light fraction or the two-fold illumination scheme with a dark interval of 30 min, 1, 1.5 and 2h. Protoporphyrin IX fluorescence kinetics were monitored during illumination. Visual skin response was monitored in the first seven days after PDT and assessed as PDT response. The PDT response decreases with decreasing length of the dark interval. Only the dark interval of 2h showed significantly more damage compared to all the other dark intervals investigated (P<0.05 compared to 1.5h and P<0.01 compared to 1h, 30 min and a single illumination). No relationship could be shown between the utilized PpIX fluorescence during the two-fold illumination and the PDT response. The rate of photobleaching was comparable for the first and the second light fraction and not dependent of the length of dark interval used. We conclude that in the skin of the hairless mouse the dark interval cannot be reduced below 2h without a significant reduction in PDT efficacy.