A circadian clock modulates the functional organization of the Japanese quail retina. Under conditions of constant darkness, rods dominate electroretinogram (ERG) b-wave responses at night, and cones dominate them during the day, yielding a circadian rhythm in retinal sensitivity and rod-cone dominance. The activity of tyrosine hydroxylase, the rate-limiting enzyme in dopamine synthesis, also exhibits a circadian rhythm in the retina with approximately threefold higher levels during the day than at night. The rhythm of tyrosine hydroxylase activity is opposite in phase to the circadian activity of tryptophan hydroxylase, the first enzyme in the melatonin biosynthetic pathway. We tested whether dopamine may be related to the physiological rhythms of the retina by examining the actions of pharmacological agents that effect dopamine receptors. We found that blocking dopamine D2 receptors in the retina during the day mimics the nighttime state by increasing the amplitude of the b-wave and shifting the retina to rod dominance. Conversely, activating D2 receptors at night mimics the daytime state by decreasing the amplitude of the b-wave and shifting the retina to cone dominance. A selective antagonist for D1 dopamine receptors has no effect on retinal sensitivity or rod-cone dominance. Reducing retinal dopamine partially abolishes rhythms in sensitivity and yields a rod-dominated retina regardless of the time of day. These results suggest that dopamine, under the control of a circadian oscillator, has a key role in modulating sensitivity and rod-cone dominance in the Japanese quail retina.

Abstract. Photoperiodic control of testis growth in Passer domesticus (house sparrow) is mediated entirely by extraretinal photoreceptors in the brain. The eyes do not participate in photoperiodically significant photoreception. Removal of the pineal organ does not affect either the response to light or, to a first approximation, the process of recrudescence. The intensity of light reaching the retina and that reaching the extraretinal photoreceptor were varied independently. This technique will make it possible to study brain photoreception in species of birds that will not tolerate blinding. Extreme caution is necessary in the interpretation of brain lesion experiments in which reproductive function is modified, since photoreception by brain receptors of unknown anatomical location affects testicular state.In an earlier paper in this series1 we reported that seemingly normal testis growth occurred in Passer domesticus exposed to inductive daylengths even though the eyes had been surgically removed. We concluded that an extraretinal photoreceptor (ERR,-extraretinal receptor for photoperiodism) must be involved in the control of testicular recrudescence and speculated on the role of retinal light perception in the intact bird. Benoit has argued that both a retinal and a brain photoreceptor are involved in the testis response of ducks.2 We discussed his arguments and concluded that the question of whether the eyes were involved at all, remained open. We have since demonstrated3 that there are no significant differences in either rate or extent of testis growth in blinded, as opposed to unoperated, sparrows held on the same lighting regimen. This result was confirmed at several different photoperiods, light intensities, and times of year; and strongly suggests, although it does not prove, that retinal light perception is not involved in photoperiodically-mediated reproductive control in P. domesticus.We have shown that the synchronization (entrainment) of the circadian rhythm of activity in the sparrow is also mediated by an extraretinal light receptor (ERR,-extraretinal receptor for entrainment).4 Further, this receptor must be in the brain, as the behavioral response to light cycles can be manipulated by affecting the amount of light that penetrates the head.5The present paper applies techniques that have been shown to affect the amount of light reaching the ERR. to the study of photoperiodically-controlled 320

No abstract

Japanese quail exhibit a robust circadian rhythm in body temperature. This rhythm is readily entrainable by 24 h light-dark (LD) cycles and persists under constant conditions. Because both the pineal organ and the eyes have been implicated as major components of the circadian system of birds, the role of these organs in generating the rhythm of body temperature was investigated. Pinealectomy, when performed alone, had little effect on the body temperature rhythm of quail either under LD or under constant darkness (DD). Most birds subjected to optic nerve section alone remained rhythmic in DD although the "robustness" of the rhythm was decreased, and 25% became arrhythmic. Birds subjected to both pinealectomy and optic nerve section behaved similarly to birds subjected to optic nerve section alone. However, complete eye removal, when performed alone or in combination with pinealectomy, caused all birds to become arrhythmic in DD. The data support the hypothesis that the eyes are the loci of circadian pacemakers in quail that act, via both neural and hormonal outputs, to preserve the integrity of (self-sustaining or damped) circadian oscillators located elsewhere.

Both light and temperature can influence the pineal's synthesis of the indoleamine melatonin. An investigation of the effects of light and temperature cycles on the pineal melatonin rhythm (PMR) showed the following: (1) Both daily light cycles and daily temperature cycles could entrain the PMR; melatonin levels peaked during the dark phase of a light-dark cycle or the cool phase of a temperature cycle. (2) The PMR could be entrained by a temperature cycle as low as 2 degrees C in amplitude in lizards held in constant light or constant darkness. (3) The length of the photoperiod or thermoperiod affected the phase, amplitude, or duration of the PMR. (4) When presented together, the effects of light and temperature cycles on the PMR depended on the phase relationship between the light and temperature cycles, as well as on the strength of the entraining stimuli, such as the amplitude of the temperature cycle. (5) Exposure to a constant cold temperature (10 degrees C) eliminated the PMR, yet a rhythm could still be expressed under a 24-hr temperature cycle (32 degrees C/10 degrees C), and the rhythm peaked during the 10 degrees C phase of the cycle. (6) A 6-hr dark pulse presented during the day did not elicit a premature rise in melatonin levels. These studies show how environmental stimuli can control the pineal rhythm of melatonin synthesis and secretion. Previous studies have supported a model in which the lizard's pineal acts as a circadian pacemaker within a multioscillator circadian system, and have implicated melatonin as a hormone by which the pineal may communicate with the rest of the system. The lizard pineal, therefore, may act as a photo- and thermoendocrine transducer translating light and temperature information into an internal cue in the form of the PMR. The PMR, in turn, may control the phase and period of circadian clocks located elsewhere, insuring that the right internal events occur at the right time of day.

All organisms exhibit significant daily rhythms in a myriad of functions from molecular levels to the level of the whole organism. Significantly, most of these rhythms will persist under constant conditions, showing that they are driven by an internal circadian clock. In birds the circadian system is composed of several interacting sites, each of which may contain a circadian clock. These sites include the pineal organ, the suprachiasmatic nucleus (SCN) of the hypothalamus, and, in some species, the eyes. Light is the most powerful entraining stimulus for circadian rhythms and, in birds, light can affect the system via three different pathways: the eyes, the pineal, and extraretinal photoreceptors located in the deep brain. Circadian pacemakers in the pineal and in the eyes of some avian species communicate with the hypothalamic pacemakers via the rhythmic synthesis and release of the hormone melatonin. Often the hypothalamic pacemakers are unable to sustain persistent rhythmicity in constant conditions in the absence of periodic melatonin input from the pineal (or eyes). It has also been proposed that pineal pacemakers may be unable to sustain rhythmicity in constant conditions without periodic neural input from the SCN. Significant variation can occur among birds in the relative roles that the pineal, the SCN, and the eyes play within the circadian system; for example, in the house sparrow pacemakers in the pineal play the predominant role, in the pigeon circadian pacemakers in both the pineal and eyes play a significant role, and in Japanese quail ocular pacemakers play the predominant role.

The site (intraocular vs. extraocular) of the biological clock driving a rhythm in melatonin content in the eyes of Japanese quail was investigated by alternately patching the left and right eyes of individual birds, otherwise held in constant light, for 12-hr periods. This patching protocol, therefore, exposed each eye to a light-dark cycle (LD 12:12) 180 degrees (12 hr) out of phase with the LD cycle experienced by the other eye. The optic nerves to both eyes were transected prior to initiating the patching protocol. The ocular melatonin rhythm (OMR) of the left eyes of quail could be entrained by this procedure 180 degrees out of phase with the rhythm expressed by the right eyes. Since optic nerve section would have deprived any putative extraocular clocks of photic entrainment information, the results show conclusively that the clock driving the OMR is located within the eye itself. In addition, the OMR of Japanese quail is remarkably unaffected by removing two potential neural inputs to the eye (sympathetic innervation from the superior cervical ganglia, and input from the isthmo-optic nucleus of the midbrain); this suggests that these inputs are not required to maintain the OMR. Finally, the clock driving the OMR of one eye does not appear to be coupled to the clock driving the OMR in the other eye, since permanently patching one eye abolished the ability of the patched eye to re-entrain to an 8-hr shift in the phase of an LD 12:12 cycle, whereas the exposed eye rapidly re-entrained to the phase-shifted cycle.

Summary. The pineal has been identified as a major circadian pacemaker within the circadian system of a number of lower vertebrates although other pacemaking sites have been implicated as well. The rhythmic synthesis and secretion of the pineal hormone, melatonin, is suggested as the mechanism by which the pineal controls circadian oscillators located elsewhere. Both light and temperature cycles can entrain the pineal melatonin rhythm. The pineal, therefore, acts as a photo and thermoendocrine transducer which functions to synchronize internal cycle with cycles in the environment. A model is presented which portrays the pineal as a major component of a 'multioscillator' circadian system and which suggests how these multiple circadian clocks are coupled to each other and to cycles of light and temperature in the external world.