Dynamics of turtle cones

The response dynamics of turtle photoreceptors (cones) were studied by the cross-correlation method using a white-noise-modulated light stimulus. Incremental responses were characterized by the kernels. White-noise-evoked responses with a peak-to-peak excursion of greater than 5 mV were linear, with mean square errors of approximately 8%, a degree of linearity comparable to the horizontal cell responses. Both a spot (0.17 mm diam) and a large field of light produced almost identical kernels. The amplitudes of receptor kernels obtained at various mean irradiances fitted approximately the Weber-Fechner relationship and the mean levels controlled both the amplitude and the response dynamics; kernels were slow and monophasic at low mean irradiance and were fast and biphasic at high mean irradiance. This is a parametric change and is a piecewise linearization. Horizontal cell kernels evoked by the small spot of light were monophasic and slower than the receptor kernels produced by the same stimulus. Larger spots of light or a steady annular illumination transformed the slow horizontal cell kernel into a fast kernel similar to those of the receptors. The slowing down of the kernel waveform was modeled by a simple low-pass circuit and the presumed feedback from horizontal cells onto cones did not appear to play a major role.     

Dynamics of chromaticity horizontal cells in the freshwater turtle retina

A model of the cone-horizontal cell circuit is presented based on morphological evidence recently found in the Reeves' turtle: a luminosity horizontal cell (LHC) that receives inputs from red-, green-, and blue-sensitive cones in the ratio of 15:3:1, a triphasic horizontal cell (THC) that receives inputs from one class of red-sensitive and from blue-sensitive cones in the ratio of 2:1; and a biphasic chromaticity horizontal cell (BHC) that receives inputs from green-sensitive cones as well as from a special class of red-sensitive (i.e. the broad spectrum) and from blue-sensitive cones in the ratio of 3:2:1. A study of the simulated impulse responses strongly suggests that the basic response patterns of the BHC and THC can be readily explained by a simple wiring diagram consisting of direct hyper-polarizing inputs from the appropriate cones and a depolarizing input from the LHC which acts as a voltage inverter. A negative feedback circuit from the LHC to the cone pedicles is included and its negative feedback gain increases as the mean illuminance level (Io) increases. The negative feedback circuit, which promotes adaptation in the cones to changing Io's, is not necessary for opponent polarization in the BHC or THC, but does explain variabilities of impulse responses.     

Light adaptation in turtle cones. Testing and analysis of a model for phototransduction.

Light adaptation in cones was characterized by measuring the changes in temporal frequency responses to sinusoidal modulation of light around various mean levels spanning a range of four log units. We have shown previously that some aspects of cone adaptation behavior can be accounted for by a biochemical kinetic model for phototransduction in which adaptation is mediated largely by a sigmoidal dependence of guanylate cyclase activity on the concentration of free cytoplasmic Ca2+, ([Ca2+]i) (Sneyd and Tranchina, 1989). Here we extend the model by incorporating electrogenic Na+/K+ exchange, and the model is put to further tests by simulating experiments in the literature. It accounts for (a) speeding up of the impulse response, transition from monophasic to biphasic waveform, and improvement in contrast sensitivity with increasing background light level, I0; (b) linearity of the response to moderate modulations around I0; (c) shift of the intensity-response function (linear vs. log coordinates) with change in I0 (Normann and Perlman, 1979); the dark-adapted curve adheres closely to the Naka-Rushton equation; (d) steepening of the sensitivity vs. I0 function with [Ca2+]i fixed at its dark level, [Ca2+]i dark; (Matthews et al., 1988, 1990); (e) steepening of the steady-state intensity-response function when [Ca2+]i is held fixed at its dark level (Matthews et al., 1988; 1990); (f) shifting of a steep template saturation curve for normalized photocurrent vs. light-step intensity when the response is measured at fixed times and [Ca2+]i is held fixed at [Ca2+]i dark (Nakatani and Yau, 1988). Furthermore, the predicted dependence of guanylate cyclase activity on [Ca2+] closely matches a cooperative inhibition equation suggested by the experimental results of Koch and Stryer (1988) on cyclase activity in bovine rods. Finally, the model predicts that some changes in response kinetics with background light will still be present, even when [Ca2+]i is held fixed at [Ca2]i dark.     

Closure of ionic channels in turtle cones

Recent observations from turtle cones on the kinetics of the light response and the variance and power spectrum of spontaneous noise are discussed in terms of the behaviour of membrane ionic channels.Presented at the EMBO-Workshop on Transduction Mechanism of Photoreceptors, Jülich, Germany, October 4–8, 1976     

Sustained feedback effects of L-horizontal cells on turtle cones

Prolonged stimulation of the periphery of their receptive field can evoke in turtle cones sustained complex depolarizations or sustained membrane oscillations. In cones in which such effects of prolonged peripheral stimulation are not apparent, the injection of short depolarizing pulses can reveal a sustained increase of electrical excitability in response to prolonged peripheral illumination. The sustained effects of prolonged peripheral illumination have characteristics similar to those of the feedback depolarizations evoked by flash peripheral stimulation: they are labile in untreated retinas, can be blocked by either hyperpolarization, Co2+ or agents that depolarize the L-horizontal cells. They are associated with a decrease in the membrane input resistance. In retinas bathed in Sr2+- or Ba2+-containing media, prolonged peripheral illumination evokes a sustained repetitive discharge of spikes. These experiments demonstrate that the feedback effects of the L-horizontal cells on the cones are not only transient but also sustained and that the sustained effects of peripheral stimulations are associated with an increase in membrane Ca2+ conductance. The possible nature of the feedback connection between L-horizontal cells and the cones is discussed.     

A molecular understanding of complementary chromatic adaptation

Photosynthetic activity and the composition of the photosynthetic apparatus are strongly regulated by environmental conditions. Some visually dramatic changes in pigmentation of cyanobacterial cells that occur during changing nutrient and light conditions reflect marked alterations in components of the major light-harvesting complex in these organisms, the phycobilisome. As noted well over 100 years ago, the pigment composition of some cyanobacteria is very sensitive to ambient wavelengths of light; this sensitivity reflects molecular changes in polypeptide constituents of the phycobilisome. The levels of different pigmented polypeptides or phycobiliproteins that become associated with the phycobilisome are adjusted to optimize absorption of excitation energy present in the environment. This process, called complementary chromatic adaptation, is controlled by a bilin-binding photoreceptor related to phytochrome of vascular plants; however, many other regulatory elements also play a role in chromatic adaptation. My perspectives and biases on the history and significance of this process are presented in this essay. This revised version was published online in August 2006 with corrections to the Cover Date.     

Characteristics and ionic processes involved in feedback spikes of turtle cones

In about 20% of the cones of untreated retinas of turtles, bright flash illumination of the periphery of their receptive field evokes a spike through the feedback mechanism from the L-horizontal cell. Such feedback spikes, never observed with central stimulation, are labile, but after they have disappeared they can be regained by depolarizing the cone. Feedback spikes are actual regenerative responses, since they show a critical threshold potential, are facilitated by cone depolarization and are blocked by hyperpolarization. They are associated with a membrane resistance decrease; tetrodotoxin (10(-5) M) does not block them. High Ca2+ media facilitate their appearance, but their effect is transient because of the cone hyperpolarization and the light response block that Ca2+ ions induce. Sr2+ ions (4-10 mM) facilitate the discharge of feedback spikes in response to peripheral illumination in every cone, whether or not it has previously shown feedback effects. In Sr2+ media, feedback spikes are stable and can be evoked by dim lights. Ba2+ (2-6 mM) also facilitates and stabilizes the discharge of feedback spikes. Co2+ and D-600 block the feedback spikes. Pharmacological agents that depolarize the L-horizontal cells, such as GABA, glutamate or nicotine, also block the feedback spikes. Both Sr2+ and Ba2+ also induce the appearance of spontaneous and off spikes, which are also blocked by Co2+, but these are not related to the feedback mechanism. These results strongly suggest that every turtle cone receives a feedback input from the L-horizontal cells, which would be able to induce an increase of the cone Ca2+ conductance, which may become regenerative.     

Dynamics of turtle horizontal cell response

The small- and large-field (cone) horizontal cells produce similar dynamic responses to a stimulus whose mean luminance is modulated by a white-noise signal. Nonlinear components increase with an increase in the mean luminance and may produce a mean square error (MSE) of up to 15%. Increases in the mean luminance of the field stimulus bring about three major changes: the incremental sensitivity defined by the amplitude of the kernels decreases in a Weber-Fechner fashion; the waveforms of the kernels are transformed from monophasic (integrating) to biphasic (differentiating); the peak response time of the kernels becomes shorter and the cells respond to much higher-frequency inputs. The dynamics of the horizontal cell response also depend on the area of the retina stimulated. Smaller spots of light produce monophasic kernels of a longer peak response time. The presence of a steady background produces three major changes in the spot kernels: the kernel's amplitude becomes larger (incremental sensitivity increases); the peak response times become shorter; the waveform of the kernels changes in a fashion similar to that observed with an increase in the mean luminance of the field stimulus. A similar enhancement in the incremental sensitivity by a steady background has also been observed in catfish, which shows that this phenomenon is a common feature of the horizontal cells in the lower vertebrate retina.     

Facultative aestivation in a tropical freshwater turtle Chelodina rugosa

• 1.1. Chelodina rugosa dug from aestivation sites at the end of the dry season were immediately alert and well coordinated.
• 2.2. Compared with non-aestivating animals, aestivating turtles had 20% higher plasma osmotic pressure and 7% higher sodium. Coupled with a small, but significant weight gain upon return to the water, this suggested the occurrence of minor dehydration in aestivating animals.
• 3.3. Plasma lactate levels of aestivating animals were low, averaging 1.99 mmol/l, consistent with aerobic rather than anaerobic metabolism having sustained their long period under ground.
• 4.4. No evidence was seen of dramatic physiological specialization. Aestivation in this species is interpreted as a primarily behavioural adaptation, made possible by typically reptilian abilities to tolerate a wide range in plasma electrolytes and to survive long periods without feeding.

     

Complementary chromatic adaptation: photoperception to gene regulation

Many photosynthetic organisms can acclimate to the quantity and quality of light present in their environment. In certain cyanobacteria the wavelengths of light in the environment control the synthesis of specific polypeptides of light harvesting antenna complex or phycobilisome. This phenomenon, called complementary chromatic adaptation, is most dramatically observed in comparison of cyanobacteria after growth in green light and red light. In red light-grown cells the phycobilisome is largely composed of phycocyanin and its associated linker polypeptides (the latter are important for the assembly of the phycocyanin subunits and their placement within the light harvesting structure); the organisms appear blue-green color. In green light-grown cells the phycobilisome is largely composed of phycoerythrin and its associated linker polypeptides; the organisms appear red in color. The ways in which these cyanobacteria sense their changing light environment and the regulatory elements involved in controlling the process of complementary chromatic adaptation are discussed in this review.     

Fine-scale thermal adaptation in a green turtle nesting population

The effect of climate warming on the reproductive success of ectothermic animals is currently a subject of major conservation concern. However, for many threatened species, we still know surprisingly little about the extent of naturally occurring adaptive variation in heat-tolerance. Here, we show that the thermal tolerances of green turtle (Chelonia mydas) embryos in a single, island-breeding population have diverged in response to the contrasting incubation temperatures of nesting beaches just a few kilometres apart. In natural nests and in a common-garden rearing experiment, the offspring of females nesting on a naturally hot (black sand) beach survived better and grew larger at hot incubation temperatures compared with the offspring of females nesting on a cooler (pale sand) beach nearby. These differences were owing to shallower thermal reaction norms in the hot beach population, rather than shifts in thermal optima, and could not be explained by egg-mediated maternal effects. Our results suggest that marine turtle nesting behaviour can drive adaptive differentiation at remarkably fine spatial scales, and have important implications for how we define conservation units for protection. In particular, previous studies may have underestimated the extent of adaptive structuring in marine turtle populations that may significantly affect their capacity to respond to environmental change.     

Phosphorelay control of phycobilisome biogenesis during complementary chromatic adaptation

For some cyanobacteria, the spectral distribution of light in the environment regulates the synthesis of specific polypeptides of the phycobilisome or light harvesting antenna complex. This process, called complementary chromatic adaptation, is controlled by a complex type of two component regulatory system. In such pathways, phosphorelay typically occurs through two histidine and two aspartate residues. Generation and complementation of mutants in CCA have uncovered three elements of this pathway, a putative sensor, RcaE, and two response regulators, RcaC and RcaF. RcaC, a large response regulator, contains two input domains, a DNA binding motif and a putative histidine phosphoacceptor domain. RcaF is a small response regulator and apparently lacks an output domain. Ordering of the pathway components has placed RcaE before RcaF, and RcaF before RcaC. This phosphorelay circuitry is novel because it has, instead of four, at least five potential phosphoacceptor domains for signal transduction.     

Biochemical bases of type IV chromatic adaptation in marine Synechococcus spp

Chromatic adaptation (CA) in cyanobacteria has provided a model system for the study of the environmental control of photophysiology for several decades. All forms of CA that have been examined so far (types II and III) involve changes in the relative contents of phycoerythrin (PE) and/or phycocyanin when cells are shifted from red to green light and vice versa. However, the chromophore compositions of these polypeptides are not altered. Some marine Synechococcus species strains, which possess two PE forms (PEI and PEII), carry out another type of CA (type IV), occurring during shifts from blue to green or white light. Two chromatically adapting strains of marine Synechococcus recently isolated from the Gulf of Mexico were utilized to elucidate the mechanism of type IV CA. During this process, no change in the relative contents of PEI and PEII was observed. Instead, the ratio of the two chromophores bound to PEII, phycourobilin and phycoerythrobilin, is high under blue light and low under white light. Mass spectroscopy analyses of isolated PEII alpha- and beta-subunits show that there is a single PEII protein type under all light climates. The CA process seems to specifically affect the chromophorylation of the PEII (and possibly PEI) alpha chain. We propose a likely process for type IV CA, which involves the enzymatic activity of one or several phycobilin lyases and/or lyase-isomerases differentially controlled by the ambient light quality. Phylogenetic analyses based on the 16S rRNA gene confirm that type IV CA is not limited to a single clade of marine Synechococcus.     

Living without oxygen: lessons from the freshwater turtle

Freshwater turtles, and specifically, painted turtles, Chrysemys picta, are the most anoxia-tolerant air-breathing vertebrates. These animals can survive experimental anoxic submergences lasting up to 5 months at 3 degrees C. Two general integrative adaptations underlie this remarkable capacity. First is a profound reduction in energy metabolism to approximately 10% of the normoxic rate at the same temperature. This is a coordinated reduction of both ATP generating mechanisms and ATP consuming pathways of the cells. Second is a defense of acid-base state in response to the extreme lactic acidosis that results from anaerobic glycolysis. Central to this defense is an exploitation of buffer reserves within the skeleton and, in particular, the turtle's shell, its most characteristic structure. Carbonates are released from bone and shell to enhance body fluid buffering of lactic acid and lactic acid moves into shell and bone where it is buffered and stored. The combination of slow metabolic rate and a large and responsive mineral reserve are key to this animal's extraordinary anaerobic capacity.     

Shedding new light on the regulation of complementary chromatic adaptation

Complementary chromatic adaptation (CCA) is a light-dependent acclimation process that occurs in cyanobacteria and likely is related to increased fitness of these organisms in natural environments. Although CCA has been studied for over 40 years, significant advances in our understanding of the molecular foundations of CCA are still emerging. In this minireview, I explore recently reported developments that include novel insights into the molecular mechanisms utilized in the photoregulation of pigmentation and the molecular basis of light-dependent changes in cellular morphology, which are central elements of the process of CCA. I also discuss future avenues of study that are expected to lead to additional progress in our understanding of CCA and our general appreciation of light sensing and photomorphogenesis in cyanobacteria.     

Action spectra for chromatic adaptation in the blue-green alga Fremyella diplosiphon

Action spectra for chromatic adaptation in Fremyella diplosiphon Drouet have been determined using techniques previously described. Action maxima are at 540 nm, with a half-band width of 80 nm, for induction of phycoerythrin synthesis (green action) and at 650 nm, with a half-band width of 90 nm, for reversal of induction of phycoerythrin synthesis (red action). The red-action spectrum includes a secondary action band centered at ca. 360 nm. Red and green action overlap from 570 to 590 nm with an isosbestic point in the vicinity of 580 nm. Shoulders are present at 520 and 630 nm. Red light is more active than green light. The 540:650-nm quantum effectiveness ratio is 1:7. There is relatively little action of either kind in the blue. The 387:540 nm and 460:650-nm quantum effectiveness ratios are zero. These results contrast strongly with previous determinations in the same organism, with major activity indicated in the blue; they are consistent with the control of photomorphogenesis in the Cyanophyta by a master pigment, analogous to phytochrome.Abbreviations APC allophycocyanin - PC physocyanin - PE phycoerythrin     

Measurement and nature of firing rate adaptation in turtle spinal neurons

There is sparse literature on the profile of action potential firing rate (spike-frequency) adaptation of vertebrate spinal motoneurons, with most of the work undertaken on cells of the adult cat and young rat. Here, we provide such information on adult turtle motoneurons and spinal ventral-horn interneurons. We compared adaptation in response to intracellular injection of 30-s, constant-current stimuli into high-threshold versus low-threshold motoneurons and spontaneously firing versus non-spontaneously-firing interneurons. The latter were shown to possess some adaptive properties that differed from those of motoneurons, including a delayed initial adaptation and more predominant reversal of adaptation attributable to plateau potentials. Issues were raised concerning the interpretation of changes in the action potentials afterhyperpolarization shape parameters throughout spike-frequency adaptation. No important differences were demonstrated in the adaptation of the two motoneuron and two interneuron groups. Each of these groups, however, was modeled by its own unique combination of action potential shape parameters for the simulation of its 30-s duration of spike-frequency adaptation. Also, for a small sample of the very highest-threshold versus lowest-threshold motoneurons, the former group had significantly more adaptation than the latter. This finding was like that shown previously for cat motoneurons supplying fast- versus slow twitch motor units.     

Dynamics of the cone-horizontal cell circuit in the turtle retina

The model of the catfish retina (Siminoff, in press) has been extended to the turtle retina with incorporation of color-coding. The turtle retina contains 6 types of cones of which 4 are red-sensitive and the other 2 are green-and blue-sensitive, respectively. The cone-horizontal circuit incorporates negative feedback from the L-HC to all the cones having input to the L-HC. By use of systems analysis, Laplace transforms and the convolution theorem, impulse responses, that give information as to gain and phase, for the cone-types and L-HC were simulated. As with the catfish retina, negative feedback gain was proportional to the dc level of the L-HC and therefore, the mean illuminance level. It was shown that this mechanism can be an important factor in chromatic adaptation, since the gains of the various cone-types are preferentially altered dependent on mean illuminance level and wavelength of the background light.