Rhythmic Production Research Articles

Ecotypic differences in rhythmicity of ethylene production in Stellaria longipes: the possible roles of ACC, MACC, and ACC oxidase

The alpine tundra ecotype of Stellaria longipes is characterized by a dwarf phenotype, whereas the prairie ecotype can be semidwarf or highly elongated depending on its environment. Related to their ability to elongate, these ecotypes also show divergent abilities to produce and respond to ethylene. The prairie ecotype produces a strong daily rhythm of ethylene, which is maintained even following stress events such as wind. The alpine ecotype exhibits a much less pronounced rhythm but greatly increases ethylene production in response to stress. We investigated what differences in ethylene synthesis might be responsible for the ability of the prairie ecotype to produce a large and regular daily rhythm of ethylene production, which in the alpine ecotype is weaker or sometimes absent. Levels of the immediate precursor to ethylene, 1-aminocyclopropane-1-carboxylate (ACC), and its major conjugate, malonyl ACC (MACC) showed no rhythm across the course of a day. Moreover ACC levels remained stable during an entire growth cycle (21 days) in the prairie ecotype, even though ethylene is known to increase especially during periods of rapid elongation. By contrast, assays of ACC oxidase performed in vivo and in vitro showed rhythms of activity similar to those of ethylene production observed in the prairie ecotype. However, the levels of ethylene produced in the ACC oxidase assays were considerably higher than levels of ethylene normally produced by unstressed plants, and the rhythm of ACC oxidase activity was observed in both ecotypes, despite the fact that alpine Stellaria longipes exhibits a less pronounced ethylene rhythm. Thus, we concluded that although ACC oxidase activity may partially account for rhythmic production of ethylene in prairie ecotypes, other controlling factors such as spatial separation of ACC from ACC oxidase should be investigated. Key words: ACC oxidase, ecotypes, ethylene, phenotypic plasticity, rhythmicity, Stellaria longipes.

Production of melatonin by the gilthead sea bream pineal: anin vivo andin vitro study

This study deals with the variations of plasma melatonin content and ofin vitro pineal melatonin secretion, in one year-old male sea bream. Plasma melatonin levels varied from almost undetectable levels during the day to 150-200 pg ml(-1) during the night, in animals maintained under a light/dark (12L/12D) cycle. The duration of the nocturnal surge corresponded to the duration of the night. A similar pattern of melatonin secretion was obtainedin vitro, with pineals cultured under a 24h L/D cycle. When the photoperiod cycle was advanced by 8h, the first nocturnal peak in melatonin secretion was more acute compared to the profile described above, but the rhythmic variations were immediately synchronized to the new L/D conditions. Under constant darkness (D/D), circadian variations in melatonin release were observed for no more than two 24h cycles. No melatonin was detected in the culture media from retinas cultured under L/D or under D/D. These results strongly suggest that the pineal is the major source of plasma melatonin in the gilthead sea bream, and that the 24h photoperiod cycle is a powerful synchronizer of the rhythmic production of melatonin by this organ.

Walking Patterns and Finger Rhythm of Older Adults

Walking patterns and rhythmic movement of the fingers were examined in a total of 1,134 male and female community residents 65 years of age and over. Walking patterns were characterized according to the ratio of step length divided by step rate (cadence), called the Walk Ratio, during level walking at preferred and maximum speeds. The walking pattern tended to change according to age; older subjects walked with shorter steps (smaller Walk Ratio). Rhythmic movement was examined using the finger-tapping test in time to the sound of a 4-Hz metronome. Hastened tapping or finger festination, in which the subject tapped faster than requested (constant error of 3 msec. and over in the intertap interval), was characteristic of aging; 16.8% of the subjects exhibited finger festination and the occurrence increased with age, especially among those in their eighties (29.3%). Finger festination was accompanied by walking patterns with an increased step rate, or a smaller Walk Ratio. These characteristics of aging were discussed as similar to extrapyramidal symptoms of walking and rhythm production in patients with Parkinson's disease.

Light-induced degradation of TIMELESS and entrainment of the Drosophila circadian clock.

Two genes, period (per) and timeless (tim), are required for production of circadian rhythms in Drosophila. The proteins encoded by these genes (PER and TIM) physically interact, and the timing of their association and nuclear localization is believed to promote cycles of per and tim transcription through an autoregulatory feedback loop. Here it is shown that TIM protein may also couple this molecular pacemaker to the environment, because TIM is rapidly degraded after exposure to light. TIM accumulated rhythmically in nuclei of eyes and in pacemaker cells of the brain. The phase of these rhythms was differentially advanced or delayed by light pulses delivered at different times of day, corresponding with phase shifts induced in the behavioral rhythms.

Positional cloning and sequence analysis of the Drosophila clock gene, timeless.

The Drosophila genes timeless (tim) and period (per) interact, and both are required for production of circadian rhythms. Here the positional cloning and sequencing of tim are reported. The tim gene encodes a previously uncharacterized protein of 1389 amino acids, and possibly another protein of 1122 amino acids. The arrhythmic mutation tim01 is a 64-base pair deletion that truncates TIM to 749 amino acids. Absence of sequence similarity to the PER dimerization motif (PAS) indicates that direct interaction between PER and TIM would require a heterotypic protein association.

Identification of rhythmic forms of speech production

An experiment was conducted to elicit stable forms of rhythm in spoken English. Stimuli were constructed, each of which consisted of a repeated series of the words take and cards. While the interval from one take to the next was a constant (1.5 s), the timing of cards relative to this interval was systematically varied. Subjects were instructed to repeat the phrase takeapackofcards, so that the relative timing of take and cards matched that of the stimulus. After some time, the stimulus was switched off, and subjects attempted to maintain the prescribed timing pattern on their own. Although there were eight target timing patterns in the stimuli, the obtained distribution of subjects’ productions was trimodal rather than octomodal. The three modes are hypothesized to correspond to three stable, preferred forms of rhythmic production for this phrase. These data strongly suggest that there is an observable rhythmic basis to English speech production. [Supported by ONR.]

Pinopsin is a chicken pineal photoreceptive molecule.

In avian pinealocytes, an environmental light signal resets the phase of the endogenous circadian pacemaker that controls the rhythmic production of melatonin. Investigation of the pineal phototransduction pathway should therefore reveal the molecular mechanism of the biological clock. The presence of rhodopsin-like photoreceptive pigment, transducin-like immunoreaction, and cyclic GMP-dependent cation-channel activity in the avian pinealocytes suggests that there is a similarity between retinal rod cells and pinealocytes in the phototransduction pathway. We have now cloned chicken pineal cDNA encoding the photoreceptive molecule, which is 43-48% identical in amino-acid sequence to vertebrate retinal opsins. Pineal opsin, produced by transfection of complementary DNA into cultured cells, was reconstituted with 11-cis-retinal, resulting in formation of a blue-sensitive pigment (lambda max approximately 470 nm). In the light of this functional evidence and because the gene is specifically expressed only in the pineal gland, we conclude that it is a pineal photosensor and name it pinopsin.

An investigation into the effects of structured music workshops with adults with mental handicap

AbstractThis study investigated the effects of structured group workshops for a population with special needs. Twenty subjects, each with a mild or moderate mental handicap, attended a series of 10 weekly 1‐hour music workshops on the structured group playing of a Javanese Gamelan. Twenty subjects formed a non‐intervention control group. The experimental hypothesis was that participation in the experimental group would produce significant improvements in musical ability as measured by the Rossi test of musical ability, devised and validated for use in this study. Significant gains in communication skills as measured by the Communication Assessment Profile for Adults with a Mental Handicap (CASP) and self‐esteem levels measured by the Khalid semantic differential technique were also postulated. It was also suggested that these gains would be significantly correlated. Results show significant gains in musical ability (instrumental rhythm production: t = 5.67, d.f. = 29, p < 0.01 and simple rhythm production: t = 8.42, d.f. = 29, p < 0.01) and communication skills (t = 4.69, d.f. = 29, p < 0.01). Moreover these results are significantly correlated (r = 0.59, p < 0.05, r = 0.75, p < 0.05, r = 0.56, p < 0.05). A ceiling effect was obtained in the measurement of self‐esteem. It is suggested that these gains derive from certain aspects of the musical communicative environment at the workshops. Suggestions for future research involve examining the possible influences on these developments.

Definition in Biology: The Case of "Bird Song"

Unrecognized different definitions for the same term can produce confusion among workers in a field. A review of over 80 definitions of bird song shows little agreement as to what defines bird song or differentiates it from calls. Defining criteria have included structural (e.g. duration), delivery (e.g. rhythmic production), physiological (e.g. hormonal control), developmental (e.g. learning), functional (e.g. territoriality), affecftive (e.g. musicality), and taxonomic (e.g. restriction to passerines) attributes of song. Each criterion has been rejected by some authors. Song definitions often are qualified with exceptions and include such qualifters as "largely arbitrary", "unclear", "not very precise", etc. Song concepts have changed over time and will continue to do so. A single song definition that covers all extant song concepts is not possible. The various song concepts serve different purposes and will be most useful if kept distinct and appropriately labeled.

The pineal organ as a component of the biological clock. Phylogenetic and ontogenetic considerations.

In conclusion, several trends are observed in regard to the phylogenetic development of the pineal organ, which are relevant for our understanding of the evolution of biological clock mechanisms. 1. The pineal organ of all vertebrates investigated thus far is capable of producing and releasing melatonin. Melatonin is rhythmically produced and released during darkness and, thus, represents an important neuroendocrine information on the ambient photoperiod. 2. The rhythmic production of melatonin is under control of endogenous oscillators and photoreceptor cells. In several nonmammalian species, these endogenous oscillators and photoreceptors are located within the pineal organ itself. In some avian species, the inherent rhythmicity of the pineal organ appears to be influenced by pacemakers located in other parts of the central nervous system. Their information may be transmitted to the pineal organ via the sympathetic innervation. This innervation develops progressively in the course of phylogeny. In mammals certain pinealocytes express proteins which are specific of retinal and pineal photoreceptors, but these proteins are obviously not involved in photoreception and phototransduction. The mammalian pineal organ lacks not only functioning photoreceptors, but also endogenous oscillators. The photoreceptor cells involved in regulation of the melatonin biosynthesis are located in the retina; the major endogenous oscillator is the suprachiasmatic nucleus (SCN) of the hypothalamus. Information from the retina and the SCN is transmitted to the mammalian pineal organ via a complex neuronal chain, whose last member is the sympathetic innervation originating from the superior cervical ganglion. This innervation is mandatory to maintain the rhythm of the melatonin biosynthesis in the mammalian pineal organ. Interestingly, the effects of noradrenaline, the major neurotransmitter in the sympathetic nerve fibers, displays opposite effects on the melatonin biosynthesis in birds and mammals: it stimulates the melatonin biosynthesis in the mammalian pineal organ, but inhibits the melatonin formation in the chicken. This conversion occurs at the level of the adrenoreceptors. 3. The intrapineal nerve cells giving rise to pinealofugal neuronal projections are reduced in the course of phylogeny. Nevertheless, direct neuronlike connections appear to exist between the pineal organ and the central nervous system of mammals. These projections originate from a population of pinealocytes. Whether such projections are involved in biological clock mechanisms remains an issue not yet resolved. The ontogenetic data reviewed support the notion that, in lower vertebrates, melatonin biosynthesis is primarily controlled by intrapineal photoreceptors, whereas, in mammals, it depends on retinal photoreceptors and the sympathetic innervation of the pineal.(ABSTRACT TRUNCATED AT 400 WORDS)

Multiple effects of an identified proprioceptor upon gastric pattern generation in spiny lobsters.

1. Using deafferented preparations of the stomatogastric nervous system of spiny lobsters (Panulirus interruptus), we stimulated the central soma of the Anterior Gastric Receptor neuron (AGR) and analyzed sensorimotor integration in the gastric central pattern generator during rhythm production. 2. Driving AGR to spike tonically at lower frequencies (10-20/s) accelerated the gastric rhythm, while higher frequencies (> or = 30/s) suppressed it. 3. Shorter spike trains in AGR evoked phase-dependent resetting of the gastric rhythm. Repetitive trains could entrain rhythms to both longer and shorter cycle periods. Some pattern-generating effects are consistent with effects upon the lateral gastric neuron, an influential member of the gastric mill network. 4. AGR affected the burst intensity of many of the gastric neurons in specific, complex ways. Some power-stroke motor neurons were excited because AGR activated excitatory, premotor interneurons (E cells). However, AGR also activated parallel, seemingly inhibitory inputs, whose mechanism remains unclear. Still other effects on motor neurons may be mediated partly by synaptic interactions within the network.

MOTOR NEURONES OF THE CRAYFISH WALKING SYSTEM POSSESS TEA+-REVEALED REGENERATIVE ELECTRICAL PROPERTIES

In crustaceans, some motor neurones (MNs) have been shown to be part of the central pattern generator in the stomatogastric system (Harris-Warrick et al. 1992; Moulins, 1990), the swimmeret system (Heitler, 1978) or the walking system (Chrachri and Clarac, 1990). These MNs induce changes in the central rhythm when depolarized and are conditional oscillators in the stomatogastric ganglion. Moreover, in the walking system, rhythmic activity can be triggered by muscarinic cholinergic agonists (Chrachri and Clarac, 1987). We have recently analyzed the role of muscarinic receptors in crayfish walking leg MNs (D. Cattaert and A. Araque, in preparation) and demonstrated that oxotremorine, a muscarinic agonist, evoked long-lasting depolarizing responses associated with an increased input resistance. The outward current blocked by oxotremorine is likely to be carried by K+, as is the case for the M current (IM) in vertebrates (Brown and Adams, 1980). In most neurones, K+ conductances play a principal role in maintaining the membrane potential at rest: for example, IM is active at the resting membrane potential, thus contributing to its maintenance, and the 'delayed-rectifier' (IK) assists the fast repolarization after an action potential. Some K+ conductances are Ca2+-dependent (IK,Ca) and are activated by an increase in internal Ca2+ concentration. In such cases, Ca2+ currents may result in hyperpolarization of the neurone through activation of IK,Ca. In opposition to these K+ currents, the direct effect of Na+ and Ca2+ conductances is to depolarize the neurone. For example, the persistant Na+ current (INap) that is responsible for the slow subthreshold depolarization termed slow pre-potentials (Gestrelius et al. 1983; Leung and Yim, 1991) participates in the formation of pacemaker depolarization (Barrio et al. 1991) and generates plateau-type responses in control conditions (Barrio et al. 1991; Llinas and Sugimori, 1980). Similarly Ca2+ or non-specific (Na+/Ca2+) conductances generate such events in Aplysia californica burster neurones (Adams and Benson, 1985), crustacean cardiac ganglion (Tazaki and Cooke, 1990), insect neurones (Hancox and Pitman, 1991) and crustacean stomatogastric ganglion (Kiehn and Harris-Warrick, 1992). Since crustacean MNs can participate in rhythm production, such depolarizing conductances may exist in most of them and may contribute to the long-lasting MN depolarizations and spike bursts present during locomotion.

Multiple circadian oscillators in the photosensitive pike pineal gland: a study using organ and cell culture.

The fish pineal organ contains typical and, in some species, modified photoreceptor cells involved in the photoperiodic control of melatonin production. In the majority of species studied, the rhythm in melatonin production is driven by an intra-pineal circadian oscillator synchronized by the light:dark cycle. In the present study, it is shown that the endogenous rhythm in melatonin release of superfused pike pineals maintained under constant darkness is expressed at temperatures of 19 degrees C, 20 degrees C, 25 degrees C, and 30 degrees C (period > 24 hr), but not at temperatures of 10 degrees C and 15 degrees C. Under constant darkness, pineal fractions containing either typical photoreceptors, modified photoreceptors, or both behaved like total organs. Dissociated pike pineal cells, cultured statically at 20 degrees C, expressed a high amplitude rhythm in melatonin secretion under a light:dark cycle. Under constant darkness, circadian oscillations, which appeared better sustained than in organ culture, were also observed. This study provides the first evidence that the rhythmic production of melatonin, by a fish pineal, is driven by a population of circadian oscillators or clocks. It is hypothesized that each typical and modified photoreceptor might be the locus of a circadian clock. Damping of the overall rhythm under constant darkness might reflect the desynchronization (uncoupling) between these clocks and/or damping of individual oscillators.

Rhythmic secretion of melatonin by the superfused pike pineal organ: thermo- and photoperiod interaction.

In the pineal organ of the pike (Esox lucius, teleost), the rhythmic production of melatonin by the photoreceptor cells is governed by a population of endogenous circadian oscillators, synchronized and entrained by the light/dark (L/D) cycle. Production of melatonin is inhibited by light and highly increased in the dark. In the present study, isolated and superfused pike pineals were exposed to a 24-hour temperature cycle of high (10 degrees C) or low (4 degrees C) amplitude, either under L/D, or under constant darkness. Under L/D, photoperiod is the dominant entraining stimulus for the melatonin secretion rhythm. It was high during the dark phase and low during the light phase, either under cold light/warm dark or under warm light/cold dark conditions. Under a warm light/cold dark cycle, the amplitude of the melatonin rhythm was reduced. In pineals cultured at 10 degrees C, a 20 degrees C temperature pulse potentiated or reduced the night-induced rise in melatonin production when applied, during the dark or during the preceding light phase, respectively. Under constant darkness, high- or low-amplitude temperature cycles could synchronize the rhythmic production of melatonin, which peaked with the high temperature. However, the shape of the oscillation could appear modified under warm subjective-L/cold subjective-D, depending on the experimental conditions. Finally, the rhythmic release of melatonin synchronized by a temperature cycle under constant darkness was no longer observed after removal of the external zeitgeber. The present study provides direct evidence that the pineal acts as a photothermotransducer.

Regulation of melatonin production by pineal photoreceptor cells: role of cyclic nucleotides in the trout (Oncorhynchus mykiss).

The light/dark cycle influences the rhythmic production of melatonin by the trout pineal organ through a modulation of the serotonin N-acetyltransferase (NAT) activity. In static organ culture, cyclic AMP (cAMP) levels (in darkness) and NAT activity (in darkness or light) were stimulated in the presence of forskolin, isobutylmethylxanthine, or theophylline. Analogues of cAMP, but not of cyclic GMP, induced an increase in NAT activity. Light, applied after dark adaptation, inhibited NAT activity. This inhibitory effect was partially prevented in the presence of drugs stimulating cAMP accumulation. In addition, cAMP accumulation and NAT activity increase, induced by forskolin, were temperature dependent. Finally, melatonin release, determined in superfused organs under normal conditions of illumination, was stimulated during the light period of a light/dark cycle by adding an analogue of cAMP or a phosphodiesterase inhibitor. However, no further increase in melatonin release was observed during the dark phase of this cycle in the presence of the drugs. This report shows for the first time that cAMP is a candidate as intracellular second messenger participating in the control of NAT activity and melatonin production by light and temperature.

A quantitative model of leaflet initiation illustrated by Murraya paniculata (Rutaceae)

A modified mathematical model based on the concept of generative centers is proposed to describe organogenesis in young leaf primordia of Murraya paniculata. Measurements of specific parameters on leaf primordia at different stages of development support the basic assumptions of the model. These assumptions are exponential elongation and widening of primordia in the organogenetic phase and rhythmic production of lateral elements at a fixed distance from the apex of the developing primordium. In general, the model provides good estimates for growth parameters such as elemental growth rates. It also provides a relatively accurate description of the shape of the primordium during the organogenesis of lateral elements or leaflet primordia. Key words: leaf development, mathematical model, organogenesis, compound leaf, Murraya paniculata.

The architecture of a dimorphic clonal fern, Lomagramma guianensis (Aublet) Ching (Dryopteridaceae)

Lomagramma guianensis possesses a dimorphic rhizome and trimorphic fronds. The juvenile genet consists of a slender, sinuous, unbranched, terrestrial rhizome which bears pinnatifid fronds. The adult rhizome form is initiated when the apex becomes scandent. It is stout, exhibiting rhythmic production of pinnate, adult-type fronds, and probably rhythmic growth. The scandent rhizome is ultimately monocarpic, producing fertile acrostichoid fronds and subsequently dying. Terrestrial runners are produced from proleptic buds at the scandent rhizome base. These are largely similar in morphology to the scandent rhizomes, but display different behaviour. The runners are relatively ephemeral structures which produce no fronds. They branch proleptically from their base to produce scandent rhizomes, and also along their length to give rise to a complex system of runners. Senescent runners which have lost contact with their parent scandent rhizome produce reiterative complexes which represent a return to the juvenile form. These reiterated rhizomes behave identically to the juvenile genets, and repeat the entire cycle of development. The architectural history of the clonal pteridophyte Lomagramma is compared to that of angiosperm trees; interesting similarities and differences are found.

Modulation of swimming rhythmicity by 5-hydroxytryptamine during post-embryonic development in Xenopus laevis.

During the first 24 h of post-embryonic development in Xenopus laevis, a rapid change in the neural activity underlying swimming occurs in which the duration of ventral root discharge on each cycle increases from a single compound impulse to discrete bursts of activity. Moreover, this change in motor output progresses rostrocaudally, suggesting that it could result from the influence of a descending neural pathway upon the spinal rhythm-generating circuitry during early post-embryonic development. To begin to examine whether serotonergic neurons of brainstem raphe nuclei might have a role in this swimming development, we have studied the effects of 5-hydroxytryptamine (5HT) on fictive swimming in embryonic and larval animals. As previously demonstrated for other vertebrate locomotor rhythms, we find that bath-applied 5HT enhances the duration of motor activity on each cycle of larval fictive swimming. In addition, our results show that the sensitivity of the swimming rhythm to exogenous 5HT follows a strict rostrocaudal gradient. In young embryos (stages 32-36) 5HT does not affect the duration of ventral root impulses per cycle; by the time of hatching (stage 37/38), rostral but not caudal discharge is enhanced, and by stage 42 (24 h post-hatching) 5HT can increase motor burst durations along most of the length of the animal. These reversible changes induced by bath-applied 5HT closely resemble the normal rostrocaudal development of burst discharge during swimming in animals some 12 h older.(ABSTRACT TRUNCATED AT 250 WORDS)

A novel T-type current underlies prolonged Ca(2+)-dependent burst firing in GABAergic neurons of rat thalamic reticular nucleus

The inhibitory GABAergic projection of thalamic nucleus reticularis (nRt) neurons onto thalamocortical relay cells (TCs) is important in generating the normal thalamocortical rhythmicity of slow wave sleep, and may be a key element in the production of abnormal rhythms associated with absence epilepsy. Both TCs and nRt cells can generate prominent Ca(2+)-dependent low-threshold spikes, which evoke bursts of Na(+)-dependent fast spikes, and are influential in rhythm generation. Substantial differences in the pattern of burst firing in TCs versus nRt neurons led us to hypothesize that there are distinct forms of transient Ca2+ current (I(T)) underlying burst discharges in these two cell types. Using whole-cell voltage-clamp recordings, we analyzed I(T) in acutely isolated TCs and nRt neurons and found three key differences in biophysical properties. (1) The transient Ca2+ current in nRt neurons inactivated much more slowly than I(T) in TCs. This slow current is thus termed I(Ts). (2) The rate of inactivation for I(Ts) was nearly voltage independent. (3) Whole-cell I(Ts) amplitude was increased when Ba2+ was substituted for Ca2+ as the charge carrier. In addition, activation kinetics were slower for I(Ts) and the activation range was depolarized compared to that for I(T). Other properties of I(Ts) and I(T) were similar, including steady-state inactivation and sensitivities to blockade by divalent cations, amiloride, and antiepileptic drugs. Our findings demonstrate that subtypes of transient Ca2+ current are present in two different classes of thalamic neurons. The properties of I(Ts) lead to generation of long-duration calcium-dependent spike bursts in nRt cells. The resultant prolonged periods of GABA release onto TCs would play a critical role in maintaining rhythmicity by inducing TC hyperpolarization and promoting generation of low-threshold calcium spikes within relay nuclei.

Pineal Photoreceptor Cells in Culture: Fine Structure, and Light/ Dark Control of Cyclic Nucleotide Levels and Melatonin Secretion

Trout pineal photoreceptor cells were dissociated by trypsin-DNase digestion and further purified by a Percoll gradient centrifugation. Total cells or purified photoreceptor cells were then embedded in a collagen gel, or layered on culture-treated polycarbonate membranes, or maintained in suspension, with RPMI 1640 medium or BGjb medium. It has been shown that cells maintain a rhythmic production of melatonin for at least seven 24 h light/dark cycles under these conditions. In this complementary study, the morphofunctional state of the photoreceptor cells was examined 1) by electron (transmission, scanning) microscopy, and 2) by pharmacological tests under different lighting conditions. Using polycarbonate membranes together with RPMI 1640 medium appeared the most suitable. The segmented organization of photoreceptor cells was well preserved when using the culture-treated membranes. It tended to disappear in cells embedded in the collagen gel and was lost after passage through the Percoll gradient. However, this one allowed obtention of an homogeneous population of photoreceptors, as recognized by their intracellular components. Intracellular organelles were rather well preserved in the embedded photoreceptors. The study also provides novel information on the nature of second messengers involved in the photoperiodic control of melatonin production in photoreceptor cells. From the effects of an adenylyl cyclase activator and a phosphodiesterase inhibitor it appeared that 1) total cells and Percoll-selected cells behaved similarly, 2) the nocturnal rise in melatonin secretion was associated with an increase in cAMP content, and 3) a fall in cAMP may be a mechanism through which light reduces melatonin secretion by photoreceptor cells. Cyclic GMP, the metabolism of which also appeared to be controlled by light, did not seem involved in the photoperiodic control of melatonin production. The method proposed herein offers interesting perspectives for the study of the photoneuroendocrine properties of isolated photoreceptor cells.