SENSORY EQUILIBRIUM AND DARK ADAPTATION IN MYA ARENARIA

1. The reaction time of Mya to light is composed of two parts. The first, a sensitization period, is an exceedingly short interval of the order of magnitude associated with photographic processes. The second is a latent period of about 1.3 seconds, during which Mya need not remain exposed to the stimulating light. 2. The process of dark adaptation in Mya is orderly. Its progress may be represented by the formation of a photosensitive substance according to the dynamics of a bimolecular reaction. See PDF for Structure 3. Photosensory equilibrium as represented by the light- and dark-adapted conditions finds a rational explanation in terms of the "stationary state" of a reversible photochemical reaction involving a photosensitive substance and its two precursors. 4. There are two corollaries to this hypothesis. The first requires that the reaction time at sensory equilibrium for a given intensity should vary inversely with the temperature; the second, that the rate of dark adaptation should vary directly with the temperature. Experiments verified both of these requirements.     

DARK ADAPTATION AND THE DARK GROWTH RESPONSE OF PHYCOMYCES

Light-adapted sporangiophores of the fungus Phycomyces respond to sudden darkening by a temporary decrease in the rate of elongation, after a latent period of several minutes. The reaction time of this "dark growth" response is compound like that of the "light growth" response. It is, moreover, shorter the more intense the previous illumination. The rate of dark adaptation following adaptation to a very large range of light intensities is found to be proportional to the logarithm of the preceding light intensity. It is shown that a constant amount of dark adaptation takes place before the response occurs. On the assumption that changes in the rate of growth reflect changes in the concentration of a substance which at constant light intensity is in equilibrium with a light-sensitive material, possible equations for such a photostationary state are examined. The most reasonable formulation requires that the partial velocity of the "light" reaction be taken proportional to log I instead of to I directly.     

THE KINETICS OF DARK ADAPTATION

1. Data are presented for the dark adaptation of four species of animals. They show that during dark adaptation the reaction time of an animal to light of constant intensity decreases at first rapidly, then slowly, until it reaches a constant minimum. 2. On the assumption that at all stages of adaptation a given response to light involves a constant photochemical effect, it is possible to describe the progress of dark adaptation by the equation of a bimolecular reaction. This supposes, therefore, that dark adaptation represents the accumulation within the sense cells of a photosensitive material formed by the chemical combination of two other substances. 3. The chemical nature of the process is further borne out by the fact that the speed of dark adaptation is affected by the temperature. The velocity constant of the bimolecular process describing dark adaptation bears in Mya a relation to the temperature such that the Arrhenius equation expresses it with considerable exactness when µ = 17,400. 4. A chemical mechanism is suggested which can account not only for the data of dark adaptation here presented, but for many other properties of the photosensory process which have already been investigated in these animals. This assumes the existence of a coupled photochemical reaction of which the secondary, "dark" reaction is catalyzed by the products of the primary photochemical reaction proper. This primary photochemical reaction itself is reversible in that its main products combine to form again the photosensitive material, whose concentration controls the behavior of the system during dark adaptation.     

THE NATURE OF FOVEAL DARK ADAPTATION

1. After a discussion of the sources of error involved in the study of dark adaptation, an apparatus and a procedure are described which avoid these errors. The method includes a control of the initial light adaptation, a record of the exact beginning of dark adaptation, and an accurate means of measuring the threshold of the fovea after different intervals in the dark. 2. The results show that dark adaptation of the eye as measured by foveal vision proceeds at a very precipitous rate during the first few seconds, that most of the adaptation takes place during the first 30 seconds, and that the process practically ceases after 10 minutes. These findings explain much of the irregularity of the older data. 3. The changes which correspond to those in the fovea alone are secured by correcting the above results in terms of the movements of the pupil during dark adaptation. 4. On the assumption that the photochemical effect of the light is a linear function of the intensity, it is shown that the dark adaptation of the fovea itself follows the course of a bimolecular reaction. This is interpreted to mean that there are two photolytic products in the fovea; that they are disappearing because they are recombining to form anew the photosensitive substance of the fovea; and that the concentration of these products of photolysis in the sense cell must be increased by a definite fraction in order to produce a visual effect. 5. It is then suggested that the basis of the initial event in foveal light perception is some mechanism that involves a reversible photochemical reaction of which the "dark" reaction is bimolecular. Dark adaptation follows the "dark" reaction; sensory equilibrium is represented by the stationary state; and light adaptation by the shifting of the stationary state to a fresh point of equilibrium toward the "dark" side of the reaction.     

THE PHOTIC SENSITIVITY OF CIONA INTESTINALIS

1. Ciona possesses two means of responding to an increase in the intensity of illumination. One is by means of a local reaction; the other is by a retraction reflex of the body as a whole. 2. The "ocelli" are not photoreceptors. The photosensitive area is in the intersiphonal region containing the neural mass. This area contains no pigment. 3. The reaction time to light is composed of a sensitization period during which Ciona must be exposed to the light, and of a latent period during which it need not be illuminated in order to react to the stimulus received during the sensitization period. 4. The duration of the reaction time varies inversely as the intensity. Analysis shows the latent period to be constant. The relation between the sensitization period and the intensity follows the Bunsen-Roscoe rule. 5. During dark adaptation the reaction time is at first large, then it decreases until a constant minimum is reached. 6. A photochemical system consisting of a reversible reaction is suggested in order to account for the phenomena observed. This system includes a photosensitive substance and its precursor, the dynamics of the reaction following closely the peculiarities of the photosensitivity of Ciona. 7. It is shown that in order to produce a reaction, a constant ratio must be reached between the amount of sensitive substance broken down by the stimulus and the amount previously broken down. 8. From the chemical system suggested certain experimental predictions were made. The actual experiments verified these predictions exactly. 9. The results obtained with regularly repeated stimulation not only fail to show any basis for a learning process or for the presence of a "higher behavior," but follow the requirements of the photochemical system suggested before.     

THE DARK ADAPTATION OF THE HUMAN EYE

During the dark adaptation of the human eye, its visual threshold decreases to a small fraction of its original value in the light. An analysis of the quantitative data describing this adaptation shows that it follows the course of a bimolecular chemical reaction. On the basis of these findings it is suggested that visual reception in dim light is conditioned by a reversible photochemical reaction involving a photosensitive substance and its two products of decomposition. Accordingly, dark adaptation depends on the course of the "dark" reaction during which the two products of decomposition reunite to synthesize the original photosensitive substance.     

Intracellular recordings from phycomyces

Intracellular recordings from the giant sporangiophore of Phycomyces stage II were obtained. The mean transmembrane potential for 30 observations was −119.9 millivolts (negative inside), and it did not change either as a result of a light stimulus or during dark adaptation. Injected depolarizing and hyperpolarizing step currents and steady currents did not produce any avidence of spike activity. We conclude that light transduction and dark adaptation in Phycomyces are not based on alterations of the transmembrane potential.     

DARK ADAPTATION IN AGRIOLIMAX

A method is described which measures the excitation of Agriolimax by light, during the progress of light adaptation, by assuming that the orientating effect of continuous excitation is expressed as a directly proportionate tension difference in the orienting muscles of the two sides of the body. The tendency toward establishment of such a tension difference is caused to work against a similar geotropic effect at right angles to the phototropic one. This enables one to study the kinetics of light adaptation, and of dark adaptation as well. The situation in the receptors is adequately described by the paradigm See PDF for Equation similar to that derived by Hecht for the differential sensitivity of various forms, but with the difference that the "dark" reaction is not only "bimolecular" but also autocatalysed by the reaction product S. The progress of dark adaptation is reflected (1) in the recovery of the amplitude of the orientation and (2) in the rates of light adaptation at different levels of the recovery; each independently supports these assumptions, for which the necessary equations have been provided. These equations also account for the relative variabilities of the angles of orientation, and, more significantly, for the two quite different kinds of curves of dark adaptation which are obtained in slightly different types of tests.     

ON "REVERSAL" OF PHOTOTROPISM IN PHYCOMYCES

Alleged reversal of the phototropism of the sporangiophores of Phycomyces by high intensities of light does not occur if infra-red radiation is properly excluded. Phototropic "indifference" alone occurs at high intensities due to equal photic action on both sides of the sporangiophore. If heat radiation is not screened out, a gradual, negative thermotropic bending takes place.     

A quantitative model of Phycomyces phototropism

A quantitative model accounting for phototropism in the wild type and in behavioural mutants of Phycomyces is described.Photomecisms (changes in the sporangiophore's growth velocity in response to changes in light intensity) are produced by a system composed of two sets of linear transducers separated by an adaptation mechanism, the first transducer being the photoreceptor.Phototropism under asymmetrical light distributions is caused by the summation of local photomecisms in the distal half of the sporangiophore, where two bright bands are produced by refraction of the incident light. The photoreceptors turn around the sporangiophore axis; they are approximately adapted to local intensity everywhere except upon entrance to the first bright band. Thus, a continuous photomecism originates at this band while the rest of the sporangiophore remains practically unstimulated.The mutants suffer a reduction in the efficiency of transduction.The behaviour of the wild type and of the mutants has been quantitatively simulated by computer. The predictions from the model fit the experimental results.     

A Kinetic Model for Adaptation and the Light Responses of Phycomyces

A kinetic model is described consisting of two sequential first order processes connected by two parallel reaction pathways, one of which is light-catalyzed. A change in light flux changes the rate constant of the light-dependent process, whereupon the levels of two chemical intermediaries readjust. The model's output duplicates all the main features of the cell's light-growth and dark-growth responses except their latent periods. An asymmetric modification of the model reproduces the two types of phototropic inversion discovered by Reichardt and Varjú and by Dennison. Simple exponential equations describe these responses of the model, as well as the theoretical course of its light and dark adaptation. It is concluded that adaptation in Phycomyces consist in the photocatalytic adjustment of the level of a metabolic reservoir.     

Effects of light and temperature on sporangiophore initiation in Pilobolus crystallinus (Wiggers) Tode

Sporangiophore initiation in Pilobolus crystallinus grown in white light was induced by either a dark or a low-temperature treatment. The period of darkness necessary to induce sporangiophore initiation was shortened by lowering the temperature. Arrhenius plots for the sporangiophore-suppressing reaction in both light and darkness consisted of two straight lines with a Q₁₀ of about 2 at lower temperatures and 8–11 at higher temperatures. The temperature at which the Q₁₀ changed was the lower, the higher the fluence rate: 14° C at 8 W/m², 19.5° C at 0.24 W/m² and 24.5° C in darkness. Possible interpretations of these results are briefly presented.Abbreviations %SP percentage of trophocysts initiating sporangiophores - D50% duration of treatment required to 50% sporangiophore initiation     

Photoperiodic time measurement in the aphid Megoura viciae

Megoura produces parthenogenetic virginoparae in long day conditions, gamic oviparae in short days. The nature of this photoperiodic response has been analysed by rearing parent apterae in a wide range of circadian and non-circadian light cycles. By varying the light and dark components independently in a two-component cycle it has been established that the time measuring function is associated primarily with the dark period. There is no evidence that an endogenous circadian oscillation is implicated: thus (a) the ‘short day’ response is abolished by ‘night interruptions’ positioned in the early or late night. But this bimodal response pattern remains unchanged when the duration of the ‘main’ photoperiod is varied from ca. 6 hr to at least 25·5 hr. The stability of the maxima within the scotophase is inconsistent with the ‘coincidence’ models of photoperiodic timing that have been proposed. It is suggested that the essential timing process operates on the hour-glass principle, beginning anew with the onset of each period of darkness; (b) night interruption experiments employing very long (up to 72 hr) scanned dark periods yielded response maxima explicable in terms of the hour-glass hypothesis but did not reveal any circadian relationship between the maxima.The ‘dark reaction’ comprises a sequence of four stages, definable by the effects of light. Stage 1, extending from dark hr 0 to ca. 2·5, is fully photoreversible: at the next dark period the entire timing sequence is repeated up to the 9·5 hr critical night length. Towards the end of stage 1 reversibility is gradually lost and after a light interruption the reaction is resumed from a later time equivalent than dark hr 0; the subsequent critical night length is therefore reduced. The extent of the photoreversal is related to light duration. The period of maximum light insensitivity (stage 2) is attained at the end of the fourth hour. From ca. dark hr 5 to just short of the critical night length light exerts an increasingly promotive action which favours the production of virginoparae. This dark process is not photoreversible. Stage 4, which begins at hr 9·5, marks the end of the timing sequence. Light will not then annul the non-promotive action of the previous long night.Light has three effects which are determined by its duration and position within the cycle. The two terminal effects, mentioned above, are associated with the interception of dark stages 1 and 3 by either short (1 hr) or longer photoperiods. Light also prepares or primes the dark period timer. Thus the critical length is increased, and timing accuracy lost, if the preceding photoperiod is less than ca. 6 hr. Light during stage 4 has a priming action but no terminal function. Repeated cycles are ‘read’ in various ways, depending on the cycle structure. For example, if light intercepts stage 3, a two-component cycle is interpreted as the overlapping sequence light/dark/light. One and the same photoperiod then acts terminally in respect of the preceding dark period and as a primer for the next dark period.There is also a mechanism for summing the promotive effects produced by repeated interruption of dark stage 3. With complex (four-component) cycles both halves of the same cycle may contribute. ‘Product accumulation’ falls below threshold if the frequency of presentation of a given promotive cycle is too low. This occurs if there are very long, relatively non-promotive dark components. Such cycles are accepted as ‘continuous darkness’.     

The sensitive period for light and temperature regulation of sporangiophore development inPhycomyces

Light and temperature markedly influence sporangiophore development inPhycomyces blakesleeanus. Under normal conditions in the dark, low temperature drastically stimulates the production of dwarf sporangiophores (microphorogenesis) and inhibits that of giant sporangiophores (macrophorogenesis). These effects of low temperature could still be observed if applied only for a short period before sporangiophore initiation. Continuous white illumination strongly inhibits microphorogenesis and slightly stimulates macrophorogenesis. Short exposures to white light noticeably inhibit microphorogenesis and stimulate macrophorogenesis when given to mycelia grown for between 90 and 160 h at 14° C or 150 h or more at 10° C. These results indicate the existence in the mycelium of developmental stages for the regulation of sporangiophorogenesis by environmental signals.     

Modulation of connexon densities in gap junctions of horizontal cell perikarya and axon terminals in fish retina: effects of light/dark cycles,interruption of the optic nerve and application of dopamine

Summary In the fish retina, connexon densities of gap junctions in the outer horizontal cells are modulated in response to different light or dark adaptation times and wavelengths. We have examined whether the connexon density is a suitable parameter of gap junction coupling under in situ conditions. Short-term light adaptation evoked low connexon densities, regardless of whether white or red light was used. Short-term dark adaptation evoked high connexon densities; this was more pronounced in the axon terminal than in perikaryal gap junctions. Under a 12 h red light/12 h dark cycle, a significant difference in connexon densities between the light and the dark period could be established in the gap junctions of the perikarya and axon terminals. Under a white light/dark cycle, only the gap junctions of axon terminals showed a significant difference. Crushing of the optic nerve resulted in an increase in connexon densities; this was more pronounced in axon terminals than in perikarya. Dopamine injected into the right eye of white-light-adapted animals had no effect. However, dopamine prevented the effect of optic-nerve crushing on connexon density. The reaction of axon-terminal gap junctions to different conditions thus resembles that of perikaryal gap junctions, but is more intense. Axon terminals are therefore thought to play an important role in the adaptation process.     

SOME EFFECTS OF DERIVATIVES OF VITAMIN K ON THE METABOLISM OF UNICELLULAR ALGAE

Vitamin K₁, 2-methyl-3-phytyl-1,4-naphthoquinone, is a substance found in all plant chloroplasts. It is, therefore, interesting to know whether it has any influence upon the metabolism of plants. Experiments made with the phytol-free derivatives like 2-methyl-1,4-naphthoquinone or the corresponding 3-oxy compound, phthiocol, gave the following results. These substances accelerate the respiration of Chlorella or Scenedesmus in a way similar to the action of the dinitrophenols. They inhibit photosynthesis and the compensation of respiration in the light strongly like hydroxylamine. In Scenedesmus they hinder the adaptation to the anaerobic utilization of hydrogen. If given after adaptation in amounts sufficient to stop photosynthesis they do not prevent photoreduction but rather stabilize this reaction against reversion. Their presence destroys the coupling between the reduction of carbon dioxide in the dark and the oxyhydrogen reaction in adapted algae. One can expect, therefore, that the natural vitamin K present in plants in concentrations of about 10^–3 M takes part in some metabolic reaction as a catalyst or regulator.     

Presence of GTP-Binding Proteins in the Plasma Membrane of the Phycomyces Sporangiophore

Ashktorab, H., and Cohen, R. J. 1994. Presence of GTP-binding proteins in the plasma membrane of the Phycomyces sporangiophore. Experimental Mycology 18, 139-149. When a plasma membrane-enriched fraction isolated from the sporangiophore of the Zygomycete Phycomyces blakesleeanus was subject to immunoblotting, two polypeptide bands reacted with an antibody directed to a conserved sequence of the ∝ subunit of G-proteins; their apparent molecular masses were 40 and 51 kDa. Upon treating the plasma membrane preparation with cholera toxin, bands at 40 and 51 kDa appeared to be ADP-ribosylated but no band appeared with pertussis toxin incubation. Apparent dissociation constants for the binding of GTPγS were determined for plasma membrane from Phycomyces sporangiophore grown in the light (K_D = 39 ± 16 nM) (±SD) and in the dark (K_D = 11 ± 6 nM). GTP served as a strong competitor for binding as did GDP, although somewhat less well, while ATP competed considerably more weakly. Northern analysis of sporangiophore mRNA displayed two bands hybridizing to the Gα2 probes coding for a G_α subunit from Dictyostelium discoideum. Furthermore, Western blotting of plasma membrane revealed several bands containing polypeptides with presumptive covalently attached immunoreactive flavins. (The prevailing evidence from the action spectra of Phycomyces is that the photoreceptor is a flavoprotein residing in the plasma membrane.) Immunoblotting also detected a H⁺ ATPase similar to the plasma membrane enzyme of yeast, corroborating our isolation of plasma membrane and suggesting another possible player in the signal responses of Phycomyces . This is apparently the first evidence for a G-protein in this class of eukaryotes. G-proteins may serve a role in the flavoprotein-mediated phototransduction system of P. blakesleeanus.     

The Duplex Nature of the Retina of the Nocturnal Gecko as Reflected in the Electroretinogram

The effect of light and dark adaptation on the electrical activity in two species of nocturnal gecko, Hemidactylus turcicus and Tarentola mauritanica was studied. The electroretinogram of both species changes from the scotopic type in the dark-adapted state to the photopic type after strong light adaptation. For the scotopic response fusion frequencies up to 18 flashes per sec. are obtained in both species. For the photopic response fusion frequencies up to 50 flashes per sec. are seen in Tarentola, and up to 25 flashes per sec. in Hemidactylus. Proceeding from dark to light adaptation the increment threshold (dI) is measured at different levels of adaptive illumination (I). At low levels of illumination the dI/I ratio is found to be small and at high levels of illumination to be large. No difference in the dI/I ratio is obtained for test lights of 462 and 605 mµ. During dark adaptation the change of threshold after exposure to moderate and weak lights (up to 10³ times dark threshold) is rather fast. After light adaptation to strong light (10⁶ times dark threshold) duplex dark adaptation curves are seen with a break separating a fast and a slow phase of dark adaptation. The significance of these results from a retina which possesses sense cells of only one type is discussed.     

DARK ADAPTATION IN DINEUTES

The level of dark adaptation of the whirligig beetle can be measured in terms of the threshold intensity calling forth a response. The course of dark adaptation was determined at levels of light adaptation of 6.5, 91.6, and 6100 foot-candles. All data can be fitted by the same curve. This indicates that dark adaptation follows parts of the same course irrespective of the level of light adaptation. The intensity of the adapting light determines the level at which dark adaptation will begin. The relation between log aI ₀ (instantaneous threshold) and log of adapting light intensity is linear over the range studied.     

An Increase in Mechanical Extensibility during the Period of Light-stimulated Growth

The sporangiophore of Phycomyces responds to a temporary increase in light intensity with a transient increase in growth rate that begins 2 to 3 minutes after the initiation of the stimulus and continues until approximately the 12th minute. Tensile tests conducted on the stage IVb sporangiophore demonstrate that an increase in mechanical extensibility of the cell wall occurs 2 minutes after the initiation of a light stimulus and continues until approximately the 15th minute. This finding supports the theory that light-stimulated plant cell expansion and rate of expansion is a function of the mechanical extensibility of the cell wall.