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The present study investigated whether room temperature (T(a)) cycles around dusk and dawn could influence the circadian rhythm of rectal temperature (T(core)), urinary 6-hydroxymelatonin sulfate during nocturnal sleep, and subjective assessments of sleep in humans. Six female and two male students served as participants. Two different T(a) conditions, cyclic and constant, were established. Two kinds of room temperature were provided to subjects: cyclic T(a) (gradual decrease from 27 degrees C to 24 degrees C between 1800 and 2200 h and gradual increase from 24 degrees C to 27 degrees C between 0300 and 0700 h) and constant T(a) (27 degrees C over 24 h). At cyclic T(a), the circadian nadir of T(core) rhythm was significantly advanced, while T(core) was significantly lower from 2300 to 0200 h and significantly higher from 0600 to 1000 h. The nocturnal concentration of 6-hydroxymelatonin sulfate in the urine during sleep was significantly higher during cyclic T(a). Waking sensation just after rising was significantly better with cyclic T(a). (Skin temperatures in the extremities-T(a)) were significantly higher with cyclic T(a) especially during the evening and night. Our results suggest that gradual change of room temperature in the evening and early morning is important in terms of sleep promotion and fresh awakening. It seems probable that mankind has been evolved to have deeper sleep under the influence of cyclic T(a) around dusk and dawn.

Effects of 17beta-estradiol and p-nonylphenol were examined in the immune and endocrine organs of Japanese quail embryos. The test compounds were injected into the yolk of embryonated eggs. Injection of estrogen resulted in (1) disappearance of lymphoid cells and flattened development of plicae in the bursa of Fabricius, (2) decreased area of thyroid follicles and height of simple cuboidal epithelial cells in the thyroid, (3) increased follicular appearance of the thymus, and (4) development of an ovotestis in male embryos. Injection of nonylphenol did not induce flattened plicae in the bursa of Fabricius or development of an ovotestis in male embryos, but it increased the disappearance of lymphoid cells from the lymphoid follicles in the bursa, decreased the height of simple cuboidal epithelial cells surrounding the thyroid follicle, and increased the follicle-like structure in the thymus in male embryos. These results suggest that nonylphenol has estrogenic effects, but these are low compared to those of estrogen itself.

We investigated the correlation between torpor frequency and capacity for non-shivering thermogenesis (NST) in Siberian hamsters (Phodopus sungorus) during 25 weeks of acclimation to cold and short days. We hypothesized that torpor use is conditioned on the development of brown adipose tissue (BAT) capacity for NST. We found that (1) the degree of noradrenaline (NA)-induced hyperthermia was positively correlated with torpor frequency and its length and depth, and (2) the maximum response to NA occurred at the time of day when hamsters naturally arouse from torpor. The present study quantifies the correlation between torpor frequency and NST capacity and we suggest that a welldeveloped NST capacity is a prerequisite for the occurrence of torpor. r 2004 Elsevier Ltd. All rights reserved.

We investigated the photoperiodic response of serotonin- and galanin (GA)- immunoreactive (ir) cells in the paraventricular organ (PVO) and infundibular nucleus (IF) of the Japanese quail and the interaction of these cells with gonadotropin-releasing hormone (GnRH)-ir neurons in the hypothalamus. Serotonin-ir cells were located in series from the PVO to the IF, and were connected with each other. The number of serotonin-ir cells differed significantly between light and dark phases on the short days (SD), but did not differ between light and dark phases on long days (LD). GA-ir cells were also found in the PVO and IF. The number of GA-ir cells under SD conditions was significantly greater than under LD conditions but did not change diurnally. Both serotonin-ir and GA-ir fibers ran along the GnRH-ir cells in the nucleus commissurae pallii. Serotonin-ir and GA-ir fibers were connected with the GnRH-ir fibers in the external layer of the median eminence (ME). We confirmed that GA-ir fibers were closely associated with serotonin-ir neurons in the PVO and IF. GA-ir neurons have at least 2 routes of regulating GnRH neurons directly, and indirectly via the serotonin-ir cells in the PVO and IF.

Extraretinal photoreceptor cells have been found in the pineal complex and deep brain of a variety of non-mammalian vertebrates. Light signals received by these photoreceptor cells seem to be a potent regulator of diverse physiological responses. Here, the pineal complex and deep brain of the Japanese grass lizard, Takydromus tachydromoides, were immunohistochemically analyzed to localize the photoreceptive molecule (opsin) and the light signal-transducing G-protein (transducin). In addition to the pineal organ and parietal eye constituting the pineal complex, we unexpectedly found a parapineal organ, which is located just below the parietal eye and is morphologically similar to the pineal organ. Both organs had photoreceptor-like cells with outer segments immunostained by anti-rhodopsin and anti-pinopsin antibodies. Neither opsin- nor transducin-like immunoreactivities were detected in the parietal eye with all the antibodies tested in this study, although its morphology resembles that of the la...

Photoperiodism and circadian rhythms have been studied intensively in birds because Aves are typical seasonal breeders and diurnal animals. Light is the most important environmental factor involved in entrainment of circadian rhythms and photoperiodism. The eyes and the extraocular photoreceptors, such as the pineal organ and hypothalamus, are reported to have an important function not only for photoreception but also for circadian organization in nonmammalian vertebrates, including birds. In this report, we review the roles of the eyes, pineal organ, and deep brain as the components of the multiphotoreceptor and multioscillator system in avian circadian organization. Microsc. Res. Tech. 53:43–47, 2001. © 2001 Wiley-Liss, Inc.

We investigated the daily rhythm of the response to noradrenaline injections in Djungarian hamsters (Phodopus sungorus sungorus) at neutral ambient temperature, under long photoperiod (L:D 12:12) and after four weeks of acclimation to cold (10°C) and short photoperiod (L:D 8:16). Animals were injected with noradrenaline (0.6 mg/kg) every four hours. Body temperature and gross motor activity were measured with MiniMitter transmitters implanted into abdominal cavity. Additionally, we measured body weight and food intake prior to, and after acclimation. After four weeks of acclimation, the experiment was performed under LD cycle and then repeated during one-day of constant light (LL) and constant darkness (DD). In animals acclimated to L:D 12:12 and ambient temperature of 25°C, noradrenaline injections caused short-lasting increase in body temperature followed by marked decrease. There was no significant difference in the magnitude of the reaction between light and dark phase of the day. After acclimation to cold and L:D 8:16, under LD conditions, we recorded significant differences between the responses to the noradrenaline injections during light and dark phase of the day. Post-injection increase was higher during the day than during the night while following noradrenaline-induced hypothermia was much more pronounced in darkness. In experiments performed after acclimation to cold and short photoperiod but during one day of LL and DD regimes, these differences were attenuated. Data presented here indicate that in cold acclimated hamsters, the response to exogenous noradrenaline depends on the time of injection and it exhibits clear daily rhythm. The rhythmicity is altered under LL and DD regimes. It seems that post-injection increase in body temperature elicits following hypothermia. This hypothermia might be of a great ecological importance. Reasonable lowering of body temperature would be a protective mechanism, allowing for energy charge restoration.

Nonmammalian vertebrates have been known to have photoreceptors in their pineal complex and deep brain as well as the retina. The extraretinal photoreceptors are believed to be essential for seasonal reproduction and regulation of circadian physiology. In this review, we discuss the relation between extraretinal photoreceptors and circadian systems in some species of nonmammalian vertebrates in terms of the pineal as the photoreceptor and oscillator for circadian rhythms, the hypothalamus as a photoreceptor and the extraretinal photoreceptors as the multi-oscillator system for circadian behavioral rhythms. Although immunohistochemical results suggest the existence of rod- and cone-type photoreceptors in the pineal, photoreception in the pineal is not always necessary for circadian systems in nonmammalian vertebrates. Circadian systems are involved in avian photoperiodic gonadal responses, and these responses are induced via deep brain photoreceptors. In some animals, deep brain photoreceptors are stained by antisera against photoreceptor proteins (rhodopsin, iodopsin and transducins). The relationship between the oscillators and photoreceptors is different among species. Physiological significance of this diversity is discussed from evolutional and developmental points of view.

Relation between retinal melatonin and corneal mitotic rhythms in the Japanese quail was investigated in experiments manipulating the ocular physiology by treatments with formoguanamine hydrochloride (FG) and eye-lid suture. In experiment 1, we investigated the effects of FG, which is known to induce photoreceptor degeneration, on retinal melatonin and corneal mitotic rhythms. FG-treatment completely abolished the retinal melatonin rhythms in both LD 12:12 and constant darkness (DD), but the corneal mitotic rhythm was maintained with high mitotic rate in darkness under a LD cycle and subjective night under DD. The result suggests that 1) the photoreceptor cells in the retina are the site for melatonin production and/or for the oscillator which drives the circadian rhythm in retinal melatonin, and 2) melatonin is not involved in generation of the corneal mitotic rhythm. In experiment 2, we investigated the effects of eye-lid suture, which is known to induce eye enlargement and bulgy cornea, on the retinal melatonin and corneal mitotic rhythms. Eye-lid suture abolished the corneal mitotic rhythm in both LD and DD, with a high mitotic rate being maintained throughout 24 hr. But retinal melatonin maintained its rhythm with high levels in darkness under a LD cycle and in subjective night under DD. The result suggests that 1) bulgy cornea in the sutured eye was induced by the increase in mitotic rate in the light period, and 2) disappearance of the corneal mitotic rhythm does not have a relation to retinal melatonin. These results suggest that retinal melatonin is not involved in generation of the corneal mitotic rhythm and that there are two circadian clock systems in the eye.

We investigated localization of visual pigments in the pineal complex of Japanese quail, Japanese grass lizards and bullfrogs immunocytochemically by use of the antiserum against bovine rhodopsin (Rh-As) and monoclonal antibodies against chicken iodopsin (Io-mAb). We also analyzed retinoids, chromophores of visual pigments, by a high performance liquid chromatography (HPLC). The outer segments and cell membranes of some photoreceptor cells in the pineal organ of the Japanese quail exhibited immunoreactivity to Rh-As, but there are also many immunonegative cells. The number of immunoreactive cells among individuals varied. Immunoreactivity to Io-mAb was weak or did not exist. The HPLC analysis revealed peaks of 11-cis and all-trans isomers of retinal in the oxime extracts of the pineal organ of Japanese quail and chickens. In the pineal of Japanese grass lizards, the outer segments of some cells were immunopositive to Io-mAb, but there were no cells immunoreactive to Rh-As. The parietal eye exhibited a well-developed lens and photoreceptor cells, but the outer segments of photoreceptor cells were immunonegative to both Rh-As and Io-mAb. In bullfrogs, three types of cells were identified in both the pineal and frontal organ; (1) immunopositive to Rh-As, (2) immunopositive to Io-mAb and (3) immunonegative to either of the antibodies. In the pineal organ of bullfrogs, 11-cis and all-trans retinal and 11-cis 3-dehydroretinal were detected, and 11-cis and all-trans retinal were also detected in the frontal organ. We detected 11-cis and all-trans retinal in the ventral part of diencephalon including the hypothalamus. Thus, the chromophore is the same between the retinal and pineal visual pigments, but the expression of opsins is different between the retina and pineal complex, which probably reflects the different function of each organ.

Breeding cycles and fecal gonadal steroids were investigated in free-living brown dippers, Cinclus pallasii. The brown dippers were marked with colored leg bands, and their behavior was observed by a field scope. The breeding season was divided into six stages according to the behavior observed; winter territory-defending, nest-building, copulation, incubation, nestling, and fledgling stages. Fecal pellets on the stones in their territories were collected and the levels of immunoreactive estradiol-17 beta (E2) and testosterone (T) in 10 mg feces were measured by radioimmunoassay. Levels of fecal E2 were high only during the copulation stage, which lasted for a few weeks, in all females of the three pairs observed. In contrast, T levels in the feces of males increased in late winter when intensive territory-defending behavior was observed, and remained high until the end of the breeding season. T levels during the nest-building stage and the copulation stage were similar to those during other nesting stages. When the birds breed only once a year, the duration of high levels of T was shorter than that of the pair with the second clutch. T levels in the feces of females showed a similar pattern to E2 levels.

We investigated the effects of castration on the development of lymphoid organs (bursa of Fabricius, thymus, and spleen) in the Japanese quail during 4–8 weeks of age under a long photoperiod (16L:8D) and the effect of testosterone implantation on the involution of the lymphoid organs under long (16L:8D) and short (8L:16D) photoperiods. Under 16L:8D, the bursa of Fabricius and thymus in intact quail grew rapidly and reached a peak at 6 weeks of age and regressed thereafter. In contrast, development of the lymphoid orgas in castrated quail was well correlated with the body growth. Testosterone treatment induced a significant reduction in relative bursal weight to body weight at 6 weeks of age under 16L:8D and 8 weeks of age under 16L:8D and 8L:16D, in relative thymus weight at 6 weeks of age under 16L:8D and 8L:16D, and in relative spleen weight at 6 weeks of age under 8L:16D. Thus, we conclude that gonadal hormones play an important role in the development and involution of lymphoid organs.

Our previous study demonstrated that the paraventricular organ (PVO) in the hypothalamus of the Japanese grass lizard (Takydromus tachydromoides) showed immunoreactivity against the light signal-transducing G-protein, transducin. This finding suggested that the PVO was a candidate for the deep-brain photoreceptor in this species. To understand functions of the PVO, we investigated distributions of transducin, serotonin, gonadotropin-releasing hormone (GnRH), and gonadotropin-inhibitory hormone (GnIH) in the lizard's brain. We immunohistochemically confirmed co-localization of transducin and serotonin in PVO neurons that showed structural characteristics of cerebrospinal fluid (CSF)-contacting neurons. GnRH-immunoreactive (ir) cells were localized in the posterior commissure and lateral hypothalamic area. Some of the serotonin-ir fibers extending from the PVO to the lateral hypothalamic area contacted the GnRH-ir cell bodies. GnIH-ir cells were localized in the nucleus accumbens, paraventricular nucleus, and upper medulla, and GnIH-ir fibers from the paraventricular nucleus contacted the lateral processes of serotonin-ir neurons in the PVO. In addition, we found that serotonin-ir fibers from the PVO extended to the suprachiasmatic nucleus (SCN), and the retrograde transport method confirmed the PVO projections to the SCN. These findings suggest that the PVO, by means of innervation mediated by serotonin, plays an important role in the regulation of pituitary function and the biological clock in the Japanese grass lizard.

Vitellogenin (VTG) produced in male fish has been used for a biomarker to study endocrine disrupters. However, the characteristics of VTG produced in male fish have not been studied well. In this study, we investigated the localization of VTG in the liver and the testis of male medaka (Oryzias latipes) treated with 17beta-estradiol (E2) and p-nonylphenol (NP). The male fish were exposed to 1 microg/L E2 and 500 microg/L NP for 1-12 days. Control groups were kept in water including only vehicle. The frozen sections of the liver and the testis were stained with immunohistochemical methods using an antiserum against medaka VTG as the first antibody. In the E2 and NP treated liver, the hepatocytes showed immunoreactivity. In particular, the cytoplasm close to the cell membrane surrounding the sinusoids was strongly immunopositive. In the testis of both treatments, the interstitial tissues and the cells (spermatocytes) in the seminiferous tubules were immunopositive. The concentration of VTG became gradually higher in both tissues with longer treatments. These results suggest that germ cells in the testis treated with E2 and NP are able to incorporate and accumulate VTG.

Up to now the regulatory mechanisms, which govern the concentrations of neurosteroids in the brain, are unclear. Seasonal breeders may serve as excellent models to understand physiological changes in neurosteroid levels and their regulatory mechanisms. The present study first investigated immunohistochemically the localization of cytochrome p450 side-chain cleavage enzyme (p450scc) and 3beta-hydroxysteroid dehydrogenase/delta(5)-delta(4)-isomerase (3betaHSD) in the brain of the newt Cynops pyrrhogaster, a seasonally breeding amphibian. Both p450scc- and 3betaHSD-like immunoreactive cells were restricted to the preoptic area. Seasonal changes in neurosteroid concentrations were then examined using adult males. Pregnenolone concentrations in the brain showed marked changes during annual breeding cycle and a maximal level in August, independent of the plasma steroid levels which were all low throughout the year. Progesterone concentrations in the brain, which were lower than pregnenolone levels, also showed peaks in April and August. In contrast, the pregnenolone sulfate level was low and its change was less pronounced. To clarify environmental factors that induce seasonal changes in neurosteroid levels, adult males were further subjected to different photoperiods and ambient temperatures for 5 weeks. Both pregnenolone and progesterone concentrations in the brain were significantly higher in the long day (LD) group than in the short day (SD) group, whereas no significant effects of different ambient temperatures on neurosteroid levels were detected. These results suggest that the newt brain possesses steroidogenic enzymes p450scc and 3betaHSD and exhibits seasonal changes in the pregnenolone and progesterone concentrations. Photoperiod may be a more important environmental factor than temperature for the regulation of neurosteroid levels in the brain.

To investigate the role of food intake in seasonal body weight changes in the Djungarian hamster, we divided animals, which had minimum body weights after 18 wk in SD, into ad-lib feeding and restricted feeding groups under a long day (LD) and a short day (SD) photoperiods. In the ad-lib feeding groups, body weight and food intake increased in parallel after 3 wk under LD and after 6–7 wk in SD because of photorefractoriness. In the restricted feeding groups, almost all animals moved around furiously (food-seeking activity) when they came to consume all the food provided at 2–3 wk of the treatment in LD and after several weeks in SD with the initiation of photorefractoriness, and ended up with death. In the restricted feeding groups, body weight increase was not observed. The furious food-seeking activity in the restricted groups and the increase of body weight specific food intake (food intake/body weight) in the ad-lib feeding groups might reflect the increased requirement for food. This increase of requirement for food seems to be one of the critical factors regulating the seasonal body weight change.

We investigated circadian ocular rhythms in the Japanese quail, Coturnix coturnix japonica. The birds were placed under light-dark cycles (LD 12∶12), constant light (LL) and constant darkness (DD), and the retinas were dissected out at four-hour intervals throughout 24 h. Following measurements were performed. (1) Melatonin content in the retina was measured by radioimmunoassay. It was low in light and several folds higher in darkness under LD 12∶12. The rhythm continued in DD, but disappeared in LL. (2) Mitotic figures in the corneal epithelium were counted. Similar rhythms to the melatonin content were observed in the corneal mitotic rate with a slight phase delay. (3) The retinas were fixed at 4-h intervals and immunostained with anti-bovine rhodopsin serum and anti-chicken iodopsin monoclonal antibodies. The outer segments of photoreceptor cells were stained intensively throughout 24 h in LD 12∶12, LL and DD. In contrast, the stainability of the locus close to the outer limiting membrane where the Golgi apparatus exists changed diurnally. Scores showing the ratio of cells with positive staining indicated high values from 4 h after the onset of light to the beginning of dark phase under LD 12∶12. The values were high throughout 24 h in LL and intermediate or low in DD. (4) To investigate the effect of melatonin on the corneal mitotic rate and visual pigments at the Golgi region, melatonin was injected into one eye and saline into the contralateral eye. Melatonin induced a phase advance in the corneal mitotic rate under LD 12∶12, but did not induce a rhythm under LL. The ratio of photoreceptor cells with positive staining to anti-visual pigment antibodies at the Golgi region was not affected by melatonin injection.

The size of the cloacal gland was found to be a reliable indicator of testicular activity of Japanese quail. Six experiments were performed to examine the effects of alternating long and short photoperiod on the size of the cloacal gland of male Japanese quail. Three types of photoperiodic cloacal responses were distinguished. Type I birds became refractory to short photoperiods after they had experienced 5 weeks or more of short days. They maintained large cloacal glands under subsequent conditions of alternating long and short photoperiod. Type II birds were intermediate between types I and III. Type III birds did not become refractory to short photoperiods after experiencing 5 weeks or more of short days. The cloacal glands responded to conditions of alternating long and short photoperiods with increases or decreases in size. Feather color on the throat was found to correspond to the type of cloacal response. Type I birds had brick-red throat feathers. Type II birds had white feathers intermingled with brick-red feathers. Type III had white throat feathers. The percentages of types I, II, and III observed in the experimental population was 67, 18, and 15%, respectively. Type III birds were used to study the effects of blinding on the cloacal response to short photoperiod. Five out of eight blinded type III birds did not lose the responsiveness to short photoperiod. These results are consistent with the view that extraocular photoreceptors participate in the photoperiodic gonadal response of Japanese quail.

The effects of artificial photoperiod and temperature on testicular FSH binding and plasma FSH levels were studied in adult male Djungarian hamsters. In Exp I, hamsters were transferred to long day (LD) photoperiods (16-h light, 8-h dark) after 8 weeks of adaptation in short day (SD) photoperiods (8-h light, 16-h dark), but the ambient temperature was maintained at 25 C throughout the experiments. A marked increase in the total FSH binding per two testes occurred between 10 and 47 days after transfer to LD, a change that was accompanied by testicular growth. Binding of FSH per testicular weight decreased during the same period. Scatchard plot analyses of the parameters indicated that LD decreased both the concentration of FSH binding sites and the equilibrium dissociation constant (Kd). Plasma FSH levels increased between 10 and 47 days after transfer to LD. In contrast, when hamsters reared under LD for 12 weeks were transferred to SD (Exp II), FSH binding per unit weight basis increased 19 weeks after transfer to SD, but the total binding per two testes decreased markedly. In Exp III, sexually mature male hamsters were subjected to different ambient temperature and photoperiods. There were no significant effects of different ambient temperatures on the testicular weight and testicular FSH binding in animals exposed for 8 weeks to either LD or SD. However, plasma FSH levels of hamsters maintained at 25 C under LD was significantly higher than FSH levels at 7 C under LD. A similar effect of temperature on plasma FSH levels was observed in hamsters under SD. The present study indicates that photoperiod is a more important environmental factor than temperature for the regulation of FSH receptor in the Djungarian hamster.

1. 1. Plasma corticosterone was determined by radioimmunoassay in 6–7-week-old male broiler type chicks, reared under several carefully controlled lighting regimes. 2. 2. When subjects were grouped by photoperiod of rearing, chicks reared in darkness had significantly lower hormone levels than diurnal controls, or than subjects reared in continuous light. 3. 3. Around-the-clock sampling revealed a diurnal corticosterone rhythm, with high daytime levels and lower night-time levels. This rhythm appeared to be retained in constant light, although phase shifted or free running. 4. 4. Neither analysis by light intensity level nor by lights on/lights off status at the time of blood sampling revealed differences in plasma corticosterone between the experimental groups which could be attributed to these factors.

Body weight, food intake, pelage color, and testis size in Djungarian hamsters are known to show annual cycles. To analyze the environmental factors that control the annual cycle of these physiological parameters, we performed an experiment with four combinations of photoperiods and temperatures (long photoperiod and high temperature, long photoperiod and low temperature, short photoperiod and high temperature, and short photoperiod and low temperature). Short photoperiod (8L16D) induced a decrease in body weight and testis size and an increase in the ratio of food storage behavior, total hair length, and the length of white distal region of hairs (a quantitative measure of pelage color) under both low (7°C) and high (25°C) temperatures. Long photoperiod (16L8D) induced the opposite effects. Low temperature induced an increase in the amount of food intake under both long and short photoperiods. under high temperature, food intake was greater in long photoperiod than in short photoperiod. Thus, we conclude that the main environmental factor for regulating annual cycles of various parameters in the Djungarian hamster is photoperiod. Photoperiodically regulated food storage behavior seems to have an important role for the strict control of body weight and food intake.

The effects of artificial photoperiod, temperature, and long-term testosterone treatment on testicular luteinizing hormone (LH) binding were studied in adult male Djungarian hamsters. In hamsters transferred to long-day (LD; 16 hr light, 8 hr dark) photoperiod 8 weeks after adaptation in short-day (SD; 8 hr light, 16 hr dark) photoperiod of 25 degrees C, testicular growth was associated with an increase in the total LH binding per two testes and a decrease in LH binding per unit testicular weight. Plasma testosterone levels reached a peak 47 days after transfer to LD and tended to decrease thereafter, while the testes continued growing. In contrast, when hamsters reared under LD conditions at 25 degrees C for 12 weeks were transferred to SD, testicular regression was associated with a decrease in plasma testosterone and the total LH binding per two testes and an increase in LH binding per unit testicular weight. A significant decrease in LH binding per unit weight compared to SD controls was observed in those hamsters exposed to SD with continuous testosterone treatment. The testosterone treatment tended to induce decrease in the total LH binding. Scatchard plot analyses of the binding suggested that changes in LH binding were due to changes in the number of binding sites. When sexually mature male hamsters were subjected for 8 weeks to two different ambient temperatures (7 degrees C and 25 degrees C) and photoperiods (LD and SD), the difference between the two temperature groups was statistically not significant regarding the weights of testes, epididymides, and prostates; plasma testosterone levels; and LH binding in either LD or SD group. These results suggest that photoperiod is a more important environmental factor than temperature for the regulation of testicular activity and LH receptors and that testosterone reduces the number of LH receptors per unit testicular weight in adult male Djungarian hamsters.

Mitotic rhythms were investigated in the chick corneal epithelium under various lighting regimens. Day‐old chicks were subjected to LD 14:10 (14 hr‐light: 10 hr‐darkness) for 2 weeks and the number of mitotic cells were counted in the central and peripheral regions of the cornea. The mitotic activity was high in darkness and low in light, and higher activity in the central region than in the peripheral region was observed. Anticipatory sharp rise in mitotic activity was observed just after light off. When chicks were reared in LD 12:12 (light on 06:00–18:00) and in inverted LD 12:12 (light on 18:00–06:00), the phase of the rhythm in normal LD 12:12 was different by 180° from that in inverted LD 12:12. When chicks were transferred to continuous light or continuous darkness after 2 weeks in LD 14:10, the mitotic rhythm disappeared in continuous light, but it continued in continuous darkness, suggesting a circadian nature of the rhythm. Two‐day old chicks, which were given only one LD cycle, also sh...

Avian gonadal activity is known to be controlled by photoperiods. In male Japa-nese quail, testicular activity is stimulated by long days and inhibited by short days. However, the photoperiodic testicular response of the quail is not completely under-stood. Thus, I investigated it under various photoperiodic conditions.When birds were reared under continuous light (LL) after hatching, they reached maturation in 6 weeks and maintained large testes as long as they were kept under LL. When birds were transferred from LL to a short photoperiod (8L : 16D), their testes showed considerable involution within 2 weeks. However, when birds received the short day treatment for 5 weeks or longer period prior to the long day treatment, their testes showed resistance to involution in the subsequent short day treatment of both young and old birds. Further, when birds were continuously kept under a short photoperiod, they showed spontaneous growth of their testes 12 weeks after hatching. Adult birds, which experienced sexual maturation under the long photoperiod, also showed spontaneous recrudescence of their testes under a short photoperiod after several weeks in the involuted state. These results clearly indicate that a short photoperiod does not always act to inhibit the testes of the Japanese quail. They also suggest that there is a kind of photorefractoriness against short days.

The influence of the photo period, of pinealectomy and of bilateral enucleation on several endocrine organ weights of Japanese quail (Coturnix cotumix japonica) have been explored. At 5 weeks after hatching, pituitary, adrenal and gonadal weights were higher, and thyroid weight was lower, in birds maintained under continuous light (24L/OD) than in those held under a short phoroperiod (8L/16D) for 3 weeks. Blinded birds did not differ from sham-pinealectomized ones in these endocrine parameters. Pinealectomy abolished the photoperiod-dependent responses of the adrenal and thyroid, while pituitary and gonadal weights were unaffected. At 6 weeks after hatching, the previously observed thyroid response was not as pronounced, and pinealectomy no longer altered the adrenal response to the photoperiod. Thus, the photoperiod and the pineal appear to be important for maturation of several endocrine glands, and the eyes do not seem to be necessary for these photo-endocrine responses in Japanese quail. (Endocrinolog...

Hypophysectomy of Japanese quailsubsequently reared under continuous light had noeffect on either pineal weight or pinealhydroxyindole-O-methyl transferase (HIOMT) activity. The gonads showed marked atrophy followinghypophysectomy. Adrenal weight was somewhatreduced but this was significant only in 10-week-oldmale quail. Thyroid weight was not affected byhypophysectomy. All hypophysectomized birdsshowed conspicuous molting. Melatonin administration (0.1, 1.0, 10, or 100µg/day for 14 days) did not affect testis weight ofadult quail under 16L/8D, 12L/12D or 8L/16Dphotoperiods. (Endocrinology 94: 1735, 1974)

Domestic chicks reared under continuous dim light developed the eye enlargement and suppressed corneal growth characteristic of light-induced avian glaucoma (LIAG) and dim light buphthalmos (DLB). Administered corticosterone moderated these effects on both the globe of the eye and the cornea. The anti-adrenal agent Metyrapone had no discernable effects on the globe parameters of the eye, although the cornea was somewhat enlarged and the anterior chamber deeper than in controls. Mitotic rate in the corneal epithelium, and adrenal and testis weight, were more sensitive indicators, showing that both the hormone and its antagonist induced changes in these experimental subjects.

We have isolated a cDNA clone encoding a deep brain photoreceptive molecule from the hypothalamic cDNA library of the toad, Bufo japonicus. The deduced amino acid sequence showed the highest similarity to that of pinopsin (75^76%) among vertebrate retinal opsins, indicating the expression of toad pinopsin in the deep brain. Antibodies raised against the C- terminal tail of toad pinopsin stained cell bodies and the knob- like structures of the cerebrospinal fluid-contacting neurons in the anterior preoptic nucleus. This region is known to play an important role in breeding behavior, suggesting that toad pinopsin acts as a photosensor for the photoperiodic gonadal response. z 1998 Federation of European Biochemical Societies.

The effects of photoperiod, temperature and testosterone treatment on plasma T3 and T4 levels were investigated in the Djungarian hamster. Plasma T3 level was affected by temperature (25 degrees C less than 7 degrees C) but not by photoperiod. Plasma T4 level was affected by photoperiod (short day less than long day) at 25 degrees C. Administration of testosterone increased plasma T4 level under short photoperiod at 25 degrees C. Thus, higher plasma T4 level under long photoperiod at 25 degrees C might be induced by testosterone.

The optimal incubation temperature for assay of the melatonin-forming enzyme, hydroxyindole-O-methyl transferase (HIOMT), of the Japanese quail pineal, was determined to be 47° C. In adult quail, HIOMT activity was high in continuous light and in the light phase of 12 L 12 D and 12 D 12 L , and was low in continuous darkness and in the dark phase of 12 L 12 D and 12 D 12 L . Age of the bird and length of the lighting treatment seemed to be important factors in demonstrating this effect of light and darkness on HIOMT activity.