Synaptic enhancement and enhanced excitability in presynaptic and postsynaptic neurons in the conditioned stimulus pathway of Hermissenda

Identified type A photoreceptors of Hermissenda express differential effects of classical conditioning. Lateral type A photoreceptors exhibit an increase in excitability to both the conditioned stimulus (CS; light) and extrinsic current. In contrast, medial type A photoreceptors do not express enhanced excitability, but do show enhancement of the medial B to medial A synaptic connection. Therefore, both enhanced excitability and changes in synaptic strength may contribute to long-term plasticity underlying classical conditioning. The activation of protein kinase C (PKC) is involved in the induction of enhanced excitability of identified type B photoreceptors produced by one-trial conditioning and the expression of enhanced excitability in B photoreceptors after multitrial classical conditioning. We have examined a possible role for persistent kinase activity in the expression of enhanced excitability in lateral type A photoreceptors and enhancement of the medial B to medial type A synaptic connection after classical conditioning. Injection of the PKC inhibitor peptide PKC(19-36) into medial type B photoreceptors of conditioned animals did not significantly change the amplitude of medial A IPSPs elicited by single spikes in the medial B photoreceptor. Injections of PKC(19-36) into medial B photoreceptors of pseudorandom controls also did not significantly change the amplitude of IPSPs recorded from the medial A photoreceptor. In contrast, spikes elicited by extrinsic current in lateral type A photoreceptors of conditioned animals were significantly reduced in frequency after intracellular injection of PKC(19-36) as compared with pseudorandom controls. Injection of the noninhibitory analog peptide [glu27]PKC(19-36) did not affect excitability. Thus, enhanced excitability in the lateral A photoreceptor of conditioned animals seems to be influenced, in part, by a constitutively active kinase or a persistent kinase activator, whereas synaptic enhancement of the connection between the medial B and medial A photoreceptors of conditioned animals may involve a different mechanism.     

Associative training results in persistent reductions in a calcium-activated potassium current in Hermissenda Type B photoreceptors.

Type B photoreceptors from the eyes of associatively trained Hermissenda had significantly greater light responses and net input resistances than Type B cells from control Ss on retention days following conditioning. Differences were still apparent when the contribution of a rapidly inactivating K+ current (IA) to the photoresponse was minimized by either depolarization-induced inactivation or block by 4-aminopyridine ions. Training-produced differences in Type B cell light responses were abolished by treatments that reduced the contribution of a calcium-activated K+ current (IK-Ca) to the light response. Under voltage-clamp conditions in which IK-Ca was isolated from other components of outward current, it was selectively reduced by associative training. The associative reduction of IK-Ca could not be attributed to training-produced reductions in the voltage-dependent calcium current of Type B cells. ICa was enhanced by associative training. A slower component of light-induced inward current was greater for Type B cells from associatively trained Ss. Previous results suggest that this slower component may be due to light-induced reduction of a calcium-activated K+ current. Training-produced reductions in a calcium-activated K+ may be responsible for enhanced photoresponses of Type B cells from conditioned Ss. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Associative learning changes intrinsic to Hermissenda Type A photoreceptors.

The eyes of the nudibranch mollusc Hermissenda contain 2 classes of photoreceptors. Type B photoreceptors exhibit increased light responses (LRs) and membrane excitability after repeated pairings of light and rotation and play an important role in the mediation of associatively produced reductions in phototactic behavior. Type A photoreceptors (TAPs) have also been shown to change with associative training. In previous research it was not possible to determine whether the effects of associative training reflected changes in synaptic input to TAPs or intrinsic changes in somatic conductances. In the present study, intracellular recordings from synaptically isolated TAPs were obtained on retention days after training, and pairing-specific decreases in light-induced generator potentials and decreases in resting input resistance were observed. Current- and voltage-clamp analysis of TAPs from untrained animals revealed that an important determinant of the steady-state LR was a calcium-activated K+ current (IK–Ca). Thus, TAPs also appear to be a primary site for associative information storage in Hermissenda. It is suggested that enhancement of IK–Ca by associative training may contribute to the diminished LR of TAPs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Contingency learning and causal detection in Hermissenda: II. Cellular mechanisms.

Neural correlates of contingency-sensitive behavioral changes in the nudibranch Hermissenda were studied by in vitro conditioning of the isolated nervous system. The additions of unpaired light-alone or hair cell-alone stimulation presentations to sequences of light-hair cell stimulation pairings were found to attenuate the cumulative depolarization of Type B photoreceptors that normally result from pairings. Unpaired light presentations produced a transitory depression of the light-induced depolarizing generator potential of the B cell. Unpaired hair cell stimulation synaptically hyperpolarized the B photoreceptor and thus acted in a retroactive manner to partially reverse the cumulative depolarization owing to prior pairings. Behavioral experiments revealed a striking temporal specificity for the decremental effects of added light and rotational stimulation. The present experiments indicate that contiguity and contingency relations are both encoded and stored in the Type B photoreceptors, and to a first approximation reflect similar biophysical mechanisms. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Preconditioning with bright light evokes a protective response against light damage in the rat retina

Constant exposure to bright light induces photoreceptor degeneration and at the same time upregulates the expression of several neurotrophic factors in the retina. At issue is whether the induced neurotrophic factors protect photoreceptors. We used a preconditioning paradigm to show that animals preconditioned with bright light became resistant to subsequent light damage. The preconditioning consisted of a 12-48 hr preexposure, followed by a 48 hr "rest phase" of normal cyclic lighting. The greatest protection was achieved by a 12 hr preexposure. Preconditioning induces a prolonged increase in two endogenous neurotrophic factors: basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). It also stimulates the phosphorylation of extracellular signal-regulated protein kinases (Erks) in both photoreceptors and Müller cells. These findings indicate that exposure to bright light initiates two opposing processes: a fast degenerative process that kills photoreceptors and a relatively slower process that leads to the protection of photoreceptors. The extent of light damage, therefore, depends on the interaction of the two processes. These results also suggest a role of endogenous bFGF and CNTF in photoreceptor protection and the importance of Erk activation in photoreceptor survival.     

Temporal order sensitivity of associative neural and behavioral changes in Hermissenda..

Hermissenda's neural and behavioral changes produced by light-rotation pairings were assessed as a function of the temporal relations between visual and vestibular stimulation. The results of in vitro simulations of conditioning indicated that simultaneous pairings (synchronous onsets and offsets of light and caudal hair cell stimulation) resulted in significantly greater cumulative depolarization of Type B photoreceptors than did either forward (light preceded hair cell stimulation) or backward (hair cell stimulation preceded light) pairings. Further experiments revealed that the attenuation of cumulative depolarization produced by the forward and backward pairings reflected the asynchrony of stimulus offsets that characterize these conditioning sequences, rather than their onsets. Analogous behavioral experiments revealed that intact animals trained with forward or backward pairings exhibited significantly less conditioning than those trained with simultaneous pairings. Strong parallels between the magnitude of cumulative depolarization from in vitro conditioning studies and the behavioral results for intact animals were also observed in experiments in which stimulus onset synchrony was held constant but offsets were made asynchronous, and vice versa. Thus Hermissenda exhibits a sensitivity to the temporal arrangement of light and rotation, and the results of behavioral conditioning can be predicted accurately from the outcome of in vitro conditioning of the isolated nervous system. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Retinal anatomy of a new species of bresiliid shrimp from a hydrothermal vent field on the Mid-Atlantic Ridge

A new species of shrimp (Rimicaris sp.) was recently collected from the Snake Pit hydrothermal vent field on the Mid-Atlantic Ridge. Until the discovery in 1989 that the deep-sea, hydrothermal vent species, Rimicaris exoculata, possessed a hypertrophied dorsal eye, everyone believed that animals recovered from vent environments were blind. Like R. exoculata, Rimicaris sp., a small orange bresiliid shrimp, has an enlarged dorsal eye specialized for detecting light in a very dim environment instead of the expected compound eye. The individual lenses characteristic of a compound eye adapted for imaging have been replaced in Rimicaris sp. by a smooth cornea underlain by a massive array of photosensitive membrane. The number of ommatidia in this species is about the same as in shrimp species that live at the surface; however, the photoreceptors are larger in the deep-sea species and the shape of the photoreceptors is markedly different. The light-sensitive region of the photoreceptor is much larger than those of other shrimp and the rest of the receptor is much smaller than normal. All screening pigment has moved out of the path of incident light to a position below the retina, and the reflecting pigment cells have adapted to form a bright white diffusing screen between and behind the photoreceptors. The ultrastructure of the microvillar array comprising the rhabdom is typical for decapod crustaceans; however, there is a much greater volume density of rhabdom (80% to 85%) than normal. There is no ultrastructural evidence for cyclic rhabdom shedding or renewal. Rimicaris sp. has apparently adapted its visual system to detect the very dim light emitted from the throats of the black smoker chimneys around which it lives.     

Biophysical and behavioral correlates of memory storage, degradation, and reactivation.

Observed neural correlates of associative memory and "forgetting" 1, 6, and 14 days after acquisition of a conditioned response (CR) in the marine snail Hermissenda. Behavioral expression of a light–rotation association, as indexed by contraction of the animal's foot in response to light, dissipated thoughout the 14-day interval such that a CR was observed 1 and 6 days after conditioning but was absent 14 days later. In relation to naive or pseudoconditioned animals, membrane resistance (inversely related to neuronal membrane conductance and directly related to excitability) of the isolated Type B photoreceptor (B cell) was elevated in conditioned animals on Days 1 and 6, whereas no elevation was detectable on Day 14. However, both the behavioral response and the elevated membrane resistance in conditioned animals were hypersensitive to light–rotation pairings (i.e., exhibited "savings") on Day 14, which is indicative of a latent memory trace. In a 2nd experiment, a current-induced depolarization of the B cell after 14 retention days resulted in an increase in input resistance of the B cell membrane in previously conditioned animals but a weaker, transient rise in resistance in B cells from animals exposed to the nonassociative control procedure 14 days earlier.… (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Structural characterization of red wine rhamnogalacturonan II

A series of experiments is described that elucidates the sources of Ca2+ that contribute to activity-dependent neuronal facilitation in Hermissenda B photoreceptors during associative conditioning. In an in vitro preparation, pairings of a 4-s light with a 3-s mechanical stimulation of presynaptic hair cells increased the input resistance and elicited spike rate (i.e., excitability) of the B photoreceptors in the Hermissenda eye, indicative of a Ca(2+)-dependent process that is analogous to associative conditioning in the intact animal. This increase in excitability was reduced but not eliminated when hyperpolarizing current was applied to the B cell during the pairings, suggesting that voltage-dependent influx of Ca2+ contributed only a portion of the total calcium signal necessary for facilitation. Moreover, no increase in excitability was observed when a comparable current-induced depolarization of the photoreceptor was substituted for light-induced depolarization. In other experiments, Ca(2+)-dependent inactivation of a light-induced Na+ current was used as an index of intracellular Ca2+ concentration. It was determined that light caused a large increase in intracellular Ca2+ concentration regardless of whether the photoreceptor was allowed to freely depolarize in response to light or was voltage clamped at its resting membrane potential. Current-induced depolarization produced a smaller increase, while presynaptic stimulation had no measurable effect. Intracellular injections of either heparin, an antagonist of intracellular Ca2+ release, or EGTA, a general Ca2+ chelator, induced comparable reductions of light-induced Ca2+ accumulation. Finally, intracellular injections of heparin blocked the pairing-induced increases in B cell excitability as effectively as injections of EGTA. Taken as a whole, these data suggest that Ca2+ release from intracellular stores may be sufficient for the induction of facilitation in this preparation, while Ca2+ influx through voltage-dependent channels may have an additive effect and provide further evidence for the ubiquitous role of Ca2+ in learning-related forms of neuronal plasticity.     

Trial-spacing effects in Hermissenda suggest contributions of associative and nonassociative cellular mechanisms.

In behaving Hermissenda, a preparatory conditioned response developed across repeated pairings of light (conditioned stimulus; CS) and rotation (unconditioned stimulus; US) with intertrial intervals (ITIs) of 60 and 120 s, but not 30 s. Likewise, contiguous in vitro stimulation of the visual and vestibular receptors, an analog of behavioral conditioning, resulted in an increase in the input resistance (i.e., excitability, a correlate of conditioning) of the B photoreceptors of the Hermissenda's eye, but only with ITIs greater than 60 s. Calcium signaling in the B cell, critical to the induction of this neuronal plasticity, was attenuated with shorter ITIs owing to (a) a reduction of the light-induced generator potential and hence voltage-dependent Ca2+ influx during the light CS, (b) a depression of the Ca2+ current that persisted throughout shorter ITIs, and (c) a steady-state inactivation of the Ca2+ current as a result of a sustained depolarization persisting from the previous trial. These results are consistent with a 2-process theory of associative learning in which a primary process (Ca2+ influx) may be opposed by a secondary process (depression of the Ca2+ current) during short ITIs. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Parietal eye-pineal morphology in lizards and its physiological implications

Pineal complexes in 85 species of lizards examined comprised seven morphological types. Members of the same family do not necessarily have the same pineal complex type. "Regressive" parietal eyes were not common except in certain arboreal lizards, primarily from the family Chameleontidae. The parietal eye is often retained in burrowing lizards, presumably because these animals are occasionally exposed to light and the parietal eye is a more suitable photoreceptor for a burrower than are lateral eyes. The pineal of certain lizards possesses a finger-like projection that extends toward the parietal eye. This extension, along with pineal wall convolutions, results in more photoreceptor cells oriented for maximal absorption of light. It is rare to find convolutions and an extension in the same pineal. Cartilage deposits and blood sinuses may modify the intensity and wavelength of light reaching the pineal. These observations suggest that the intracranial pineal of lizards is a more important photoreceptor than was previously realized, a situation that may be a factor in the occasional "failure" of parietalectomy experiments.     

Implicating causal relations between cellular function and learning behavior.

Learning in the nudibranch mollusc Hermissenda shows many features of vertebrate associative conditioning. Several of these behaviors are reviewed to show how the demonstrated close temporal correspondence of cellular and behavioral functions further implicates certain causal relations. Studies of the shadow withdrawal behavior of Hermissenda suggest a causal relation between the long-lasting depolarization of the Type B photoreceptor and the animal's reduced ability to turn toward the light at light/dark boundaries. Whereas the shadow response corresponded to cellular events at the end of a light step, responses to the onset of light or rotation were largely unexplored. By using a different approach, we identified behavioral responses during the first few seconds of stimulation with light and rotation. These responses, for which Pavlovian conditioning was demonstrated, correspond closely in time to known cellular correlates. Consideration of the known neural pathways that can control classically conditioned foot-shortening and shadow withdrawal leads to the suggestion that some effects of learning operate through different neural pathways but these different pathways may be modulated by the same primary changes in the Type B photoreceptor. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Circadian clock functions localized in xenopus retinal photoreceptors

A circadian oscillator that regulates visual function is located somewhere within the vertebrate eye. To determine whether circadian rhythmicity is generated by retinal photoreceptors, we isolated and cultured photoreceptor layers from Xenopus retina. On average, 94% of the viable cells in these preparations were rod or cone photoreceptors. Photoreceptor layers produced melatonin rhythmically, with an average period of 24.3 hr, in constant darkness. The phase of the melatonin rhythm was reset by in vitro exposure of the photoreceptor layers to cycles of either light or quinpirole, a D2 dopamine receptor agonist. These data indicate that other parts of the eye are not necessary for generation or entrainment of retinal circadian melatonin rhythms and suggest that rod and/or cone photoreceptors are circadian clock cells.     

Uptake of precursor and synthesis of transmitter in a histaminergic photoreceptor

As a first step in understanding how the supply of the neurotransmitter histamine is maintained in a photoreceptor, we followed the uptake and metabolism of the immediate precursor of histamine, histidine. [3H]Histidine taken up into photoreceptors and glia was detected using autoradiography, and synthesis of [3H]histamine from [3H]histidine was assayed with thin-layer chromatography. Photoreceptors from barnacles were pulsed (15 min) with [3H]histidine (0.2-200 microM), then maintained in normal saline for up to 24 hr. Autoradiography showed that photoreceptor somata, axons, and presynaptic arbors were labeled, but only weakly, like (nonhistaminergic) ganglion cells. Label instead was concentrated over surrounding glia. Stimulating preparations with light did not increase photoreceptor labeling. Grain counts from photoreceptor axons showed uptake of [3H]histidine into these neurons by a Na+-dependent mechanism with a Km of approximately 50 microM. Over 24 hr only 1% of the [3H]histidine taken up by preparations was converted to [3H]histamine either in the dark or in the light. Injections of [3H]histidine directly into photoreceptors established that synthesis takes place within the photoreceptors and confirmed that stimulation with light did not measurably affect the rate of conversion of [3H]histidine to [3H]histamine. These results suggest that de novo synthesis of transmitter is unlikely to be as important as its reuptake in maintaining neurotransmitter supply in these photoreceptor terminals. In support of this conclusion, photoreceptors accumulated more label when transmitter release was stimulated with high K+ and histamine uptake was antagonized with chlorpromazine.     

Rhythmic activities of isolated and clustered pacemaker neurons and photoreceptors of Aplysia retina in culture

Each eye of Aplysia contains a circadian clock that produces a robust rhythm of optic nerve impulse activity. To isolate the pacemaker neurons and photoreceptors of the eye and determine their participation in the circadian clock and its generation of rhythmic autoactivity, the retina was dissociated and its cells were placed in primary cell culture. The isolated neurons and photoreceptors survived and vigorously extended neurites tipped with growth cones. Many of the photoreceptors previously described from histological sections of the intact retina were identified in culture, including the large R-type photoreceptor, which gave robust photoresponses, and the smaller tufted, whorled, and flared photoreceptors. The pacemaker neurons responsible for the rhythmic impulse activity generated by the eye were identified by their distinctive monopolar morphology and recordings were made of their activity. Isolated pacemaker neurons produced spontaneous action potentials in darkness, and pacemaker neurons attached to fragments of retina or in an isolated cluster interacted to produce robust spontaneous activity. This study establishes that isolated retinal pacemaker neurons retain their innate autoactivity and ability to produce action potentials in culture and that clusters of coupled pacemaker neurons are capable of generating robust autoactivity comparable to pacemaker neuron rhythmic activity recorded in the intact retina, which was previously shown to correspond to 1:1 with the optic nerve compound action potential activity.     

Saccular influence on the otolith-spinal reflex and posture during sudden falls of the cat

We made intradendritic recordings in Purkinje cells (n = 164) from parasaggital slices of cerebellar lobule HVI obtained from rabbits given paired presentations of tone and periorbital electrical stimulation (classical conditioning, n = 27) or explicitly unpaired presentations of tone and periorbital stimulation (control, n = 16). Purkinje cell dendritic membrane excitability, assessed by the current required to elicit local dendritic calcium spikes, increased significantly in slices from animals that received classical conditioning. In contrast, membrane potential, input resistance, and amplitude of somatic and dendritic spikes were not different in slices from animals given paired or explicitly unpaired stimulus presentations. The location of cells with low thresholds for local dendritic calcium spikes suggested that there are specific sites for learning-related changes within lobule HVI. These areas may correspond to learning "microzones" and are consistent with locations of learning-related in vivo changes in Purkinje cell activity. Application of 4-aminopyridine, an antagonist of the rapidly inactivating potassium current IA, reduced the threshold for dendritic spikes in slices from naive animals to levels found in slices from trained animals. In cells where thresholds for eliciting parallel fiber-stimulated Purkinje cell excitatory postsynaptic potentials (EPSPs) were measured, levels of parallel fiber stimulation required to elicit a 6-mV EPSP as well as a 4-mV EPSP (n = 30) and a Purkinje cell spike (n = 56) were found to be significantly lower in slices from paired animals than unpaired controls. A classical conditioning procedure was simulated in slices of lobule HVI by pairing a brief, high-frequency train of parallel fiber stimulation (8 pulses, 100 Hz) with a brief, lower frequency train of climbing fiber stimulation (3 pulses, 20 Hz) to the same Purkinje cell. Following paired stimulation of the parallel and climbing fibers, Purkinje cell EPSPs underwent a long-term (> 20 min) reduction in peak amplitude (-24%) in cells (n = 12) from animals given unpaired stimulus presentations but to a far less extent (-9%) in cells (n = 20) from animals given in vivo paired training. Whereas 92% of cells from unpaired animals showed pairing-specific depression, 50% of cells from paired animals showed no depression and in several cases showed potentiation. Our data establish that there are localized learning-specific changes in membrane and synaptic excitability of Purkinje cells in rabbit lobule HVI that can be detected in slices 24 h after classical conditioning. Long-term changes within Purkinje cells that effect this enhanced excitability may occlude pairing-specific long-term depression.     

A myosin III from Limulus eyes is a clock-regulated phosphoprotein

The lateral eyes of the horseshoe crab Limulus polyphemus undergo dramatic daily changes in structure and function that lead to enhanced retinal sensitivity and responsiveness to light at night. These changes are controlled by a circadian neural input that alters photoreceptor and pigment cell shape, pigment migration, and phototransduction. Clock input to the eyes also regulates photomechanical movements within photoreceptors, including membrane shedding. The biochemical mechanisms underlying these diverse effects of the clock on the retina are unknown, but a major biochemical consequence of activating clock input to the eyes is a rise in the concentration of cAMP in photoreceptors and the phosphorylation of a 122 kDa visual system-specific protein. We have cloned and sequenced cDNA encoding the clock-regulated 122 kDa phosphoprotein and show here that it is a new member of the myosin III family. We report that Limulus myosin III is similar to other unconventional myosins in that it binds to calmodulin in the absence of Ca2+; it is novel in that it is phosphorylated within its myosin globular head, probably by cAMP-dependent protein kinase. The protein is present throughout the photoreceptor, including the region occupied by the photosensitive rhabdom. We propose that the phosphorylation of Limulus myosin III is involved in one or more of the structural and functional changes that occur in Limulus eyes in response to clock input.     

Survival of purified rat photoreceptors in vitro is stimulated directly by fibroblast growth factor-2

Basic fibroblast growth factor (FGF-2) influences the differentiation and survival of retinal photoreceptors in vivo and in vitro, but it is not known whether it acts directly on photoreceptor FGF receptors or indirectly through activation of surrounding cells. To clarify the effects of FGF-2 on photoreceptor survival, we developed a purified photoreceptor culture system. The outer nuclear layers of postnatal day 5-15 rat retinas were isolated by vibratome sectioning, and the photoreceptor fractions obtained were enzymatically dissociated. Photoreceptors were maintained in monolayer culture for 1 week in a chemically defined medium. Immunocytochemical labeling showed that >99.5% of cells were photoreceptors, and glial contamination represented approximately 0. 2%. Photoreceptors from postnatal day 5-9 retinas survived for at least 24 hr in vitro, whereas cells from postnatal day 10-15 retinas died rapidly. Subsequent studies performed with postnatal day 5 photoreceptors showed that their survival was increased in a dose-dependent manner after the addition of FGF-2. In control cultures, 36% of originally seeded photoreceptors were alive after 5 d in vitro, and in the presence of 20 ng/ml FGF-2 this number was doubled to 62%. This increase was not caused by proliferation of photoreceptor precursors. Denaturing or blocking FGF-2 prevented enhancement of survival. Conversely, only 25.5% of photoreceptors survived in the presence of epidermal growth factor (EGF). FGF- and EGF-receptor mRNA and proteins were detected in purified photoreceptors in vitro, and addition of FGF-2 or EGF led to tyrosine phosphorylation of photoreceptor proteins. These data support a direct mechanism of action for FGF-2 stimulation of photoreceptor survival.     

Modulation of ion channels in rod photoreceptors by nitric oxide

Subcellular compartments in the outer retina of the larval tiger salamander were identified as likely sites of production of nitric oxide (NO), a recently recognized intercellular messenger. NADPH diaphorase histochemistry and NO synthase immunocytochemistry labeled photoreceptor ellipsoids and the distal regions of bipolar and glial cells apposing photoreceptor inner segments, suggesting a role for NO in visual processing in the outer retina. We investigated the actions of NO on several rod photoreceptor ion channels. Application of the NO-generating compound S-nitrosocysteine increased Ca2+ channel current and a voltage-independent conductance, but had no affect on voltage-gated K+ or nonspecific cation currents. Given the steep relation between voltage-dependent Ca2+ influx and photoreceptor synaptic output, these results indicate that NO could modulate transmission of the photoresponse to second order cells.     

Phytochromes and cryptochromes in the entrainment of the Arabidopsis circadian clock

Circadian clocks are synchronized by environmental cues such as light. Photoreceptor-deficient Arabidopsis thaliana mutants were used to measure the effect of light fluence rate on circadian period in plants. Phytochrome B is the primary high-intensity red light photoreceptor for circadian control, and phytochrome A acts under low-intensity red light. Cryptochrome 1 and phytochrome A both act to transmit low-fluence blue light to the clock. Cryptochrome 1 mediates high-intensity blue light signals for period length control. The presence of cryptochromes in both plants and animals suggests that circadian input pathways have been conserved throughout evolution.