Role of melatonin in photoperiodic time measurement in the migratory redheaded bunting (Emberiza bruniceps) and the nonmigratory Indian weaver bird (Ploceus philippinus)

In the present study, we asked the question whether physiological responses to day length of migratory redheaded bunting (Emberiza bruniceps) and nonmigratory Indian weaver bird (Ploceus philippinus) are mediated by the daily rhythm of melatonin. Melatonin was given either by injection at certain times of the day or as an implant. In series I experiments on the redheaded bunting, melatonin was administered by subcutaneous injections daily at zeitgeber time (ZT) 4 (morning) or ZT10 (evening) and by silastic capsules in photosensitive unstimulated buntings that were held in natural day lengths (NDL) at 27 degrees N beginning from mid February, and in artificial day lengths (ADL, 12L:12D and 14L:10D). Melatonin did not affect the photoperiod-induced cycles of gain and loss in body mass and testicular growth-involution, but there was an effect on temporal phasing of the growth-involution cycle of testes in some groups. For example, the rate of testicular growth and development was faster in birds that received melatonin injection at ZT4 in NDL, and was slower in birds that carried melatonin implants both in NDL and ADL. In series II experiments on Indian weaver birds, melatonin was given in silastic capsules in the first week of September when they still had large gonads. Birds were exposed for 12 weeks to short day length (8L:16D; group 1), to long day length (eight weeks of 16L:8D and four weeks of 18L:6D; group 2), or to both short and long day lengths (four weeks each of 8L:16D, 16L:8D, and 18L:6D; groups 3 and 4). Whereas groups 1 to 3 carried melatonin or empty implant from the beginning, group 4 received one after four weeks. All birds underwent testicular regression during the first four weeks irrespective of the photoperiod they were exposed to or the implant they carried in, and there was a slight re-initiation of testis growth in some birds during the next eight weeks of long day lengths. However, with the exception of group 2, there was no difference in mean testis volume during the period of experiment between the melatonin- and empty-implant birds. The data on androgen-dependent beak color also supported the observations on testes. Together, these results do not support the idea that the daily rhythm of melatonin is involved in the photoperiodic time measurement in birds. However, there may still be a role of melatonin in temporal phasing of the annual reproductive cycle in birds.     

Effect of testosterone on testes, body weight and plumage regeneration in photorefractory male redheaded bunting Emberiza bruniceps

Various doses of testosterone propionate (TP) were administered for 10 days to refractory male redheaded buntings exposed to 15L:9D in last week of June, shortly before the onset of molt, to determine the effect of exogenous male hormone on the testes, body weight and plumage regeneration in photorefractory birds. While 5-, 10-, or 25 micrograms of TP bird-1 day-1 did not affect the testes, body weight or feather regeneration, testicular growth was stimulated but body weight declined and plumage regeneration prevented in birds that received 150- or 250 micrograms of TP bird-1 day-1. In contrast, a 50 micrograms treatment, although unable to induce the growth of the testes, caused loss in body weight and feather regeneration. Also, a 25 micrograms dose of TP administered to photostimulated birds induced complete testicular atrophy. These results suggests that testosterone exerts dose-dependent effects on the testes, body weight and plumage regeneration in buntings. A positive correlation is found between the amount of hormone and the testes growth, while the increasing amount of hormone has negative effect on the body weight and regeneration of feathers.     

Biological clocks help reduce the physiological conflicts in avian migrants

We present evidence from experiments on overwintering populations of two Palearctic-Indian latitudinal migratory birds, the black-headed bunting (Emberiza melanocephala) and the red-headed bunting (E. bruniceps), that the bird’s clock in interaction with day length regulates seasonal rhythms of migration and reproduction such that physiological conflict between them is reduced. Initiation and termination of the body mass and testicular cycles are separately regulated photoperiodic events. For example, under stimulatory photoperiods the response curve of body mass does not overlap with that of the testicular growth. A response-specific photoperiodism is adaptive, since gain in body mass, critical to spring migration, precedes gonadal recrudescence. Finally, migration as indicated by the night-time migratory restlessness under experimental situations (e.g., intense locomotion under caged condition-called Zugunruhe) appears to be regulated by a separate circadian oscillator.     

The thyroid and photoperiodic control of seasonal reproduction in American tree sparrows (Spizella arborea)

To explore the role of the thyroid gland in the control of seasonal reproduction in obligately photoperiodic American tree sparrows (Spizella arborea), the effects of (1) thyroxine administered in drinking water to thyroid-intact photosensitive or photorefractory birds, and (2) radiothyroidectomy before and after photostimulation and during photorefractoriness were examined. Chronic administration of pharmacological doses of thyroxine induced testicular growth and usually regression in initially photosensitive birds held on short or intermediate daylengths. Some thyroxine-treated birds with regressed testes were absolutely photorefractory, but most remained photosensitive. Exogenous thyroxine never induced testicular growth in photorefractory birds moved to short days, though it often impeded, and sometimes even blocked, the recovery of photosensitivity. Although circumstantial, these effects of exogenous thyroxine are consistent with an hypothesis that assigns to thyroid hormones two roles — one stimulatory and the other inhibitory — in the control of seasonal reproduction. Radiothyroidectomy before photostimulation inhibited (but did not prevent) photoinduced testicular growth, blocked spontaneous testicular regression, suppressed molt, and prevented photorefractoriness. Moreover, as demonstrated by testicular growth after thyroxine replacemnt therapy, radiothyroidectomy during photorefractoriness later restored photosensitivity despite continued photostimulation. Thus, euthyroidism is an essential condition for maximizing (but not for initiating) photoinduced testicular growth and for triggering and maintaining photorefractoriness in photostimulated tree sparrows. However, when performed early during photostimulation, radiothyroidectomy neither immediately induced nor later blocked spontaneous testicular regression. Thus, endogenous thyroid hormones and long days may interact during a critical period to program a sequence of physiological events that plays out as photorefractoriness in chronically photostimulated birds. Such an organizational event cannot be permanent, for seasonal reproduction is episodic and its control mechanism necessarily cyclic. Because thyroidectomy simulated the well-known restorative effect of short days (and exogenous thyroxine impeded it), short days may dissipate photorefractoriness by creating a milieu wherein thyroid hormones are deficient or inactive.Abbreviations ANOVA analysis of variance - bTSH bovine thyroid stimulating hormone - GnRH gonadotropin-releasing hormone - LH luteinizing hormone - nL: nD daily light: dark regime (n is duration in hours) - SEM standard error of the mean - SNK Student-Newman-Keuls test - T₄ thyroxine - TH thyroid hormone - TR thyroid hormone receptor     

The effects of combined serotonin and dopamine precursor application during testicular photosensitivity vs photorefractoriness of the red headed bunting (emberiza bruniceps)

Abstract

A study of the temporal synergism of serotonergic and dopaminergic activity in the regulation of reproductive seasonality of photosensitive and photorefractory migratory Red headed bunting was undertaken. In experiment 1, groups of birds kept in natural day length (NDL) in their photosensitive phase (April) received daily injections of L‐DOPA (L‐dihydroxyphenylalanine, a dopamine precursor) at 8 and 12 hr after 5‐HTP (5 hydroxytryptophan, a serotonin precursor) administration. One control group received two daily injections of saline. The injections with 12‐hr temporal relation induced early recrudescence and full gonadal growth was achieved much in advance to that of control birds. The applications with 8‐hr temporal relation suppressed the testicular growth initially but later on this effect was eliminated by NDL of May‐June. In Experiment 2, photorefractory birds also received the treatments but the temporal relation of 5‐HTP and L‐DOPA had no effect on the regressed gonads of these birds.

This study indicates that injections with specific phase relations of dopamine and serotonin precursors may alter seasonal reproductive conditions only in the photosensitive phase but had no effect in photorefractory birds.     

Thyroidectomy results in termination of photorefractoriness in starlings (Sturnus vulgaris) kept in long daylengths

Four groups of 10 male starlings were transferred from short daylengths (8 h light/day) to long daylengths (18 h light/day), which caused the tests to develop rapidly to maximum size and then to decrease to minimal size as birds became photorefractory. Birds were surgically thyroidectomized at 8, 16 or 28 weeks. A fourth group was left intact. Testicular volume and plasma FSH and prolactin concentrations were measured. After 42 weeks all birds were castrated and plasma FSH was measured during the next 6 weeks. Testicular growth began in all thyroidectomized birds between 4 and 8 weeks after thyroidectomy. By 42 weeks, the testes of all thyroidectomized birds were large, whereas those of intact birds were still of minimal size. Plasma FSH concentrations remained low in all birds and plasma prolactin values, originally elevated by long daylengths, decreased at a similar rate in thyroidectomized and intact birds. After castration at 42 weeks, plasma FSH values increased rapidly in all thyroidectomized birds but remained low in non-thyroidectomized birds. The results demonstrate that thyroidectomy of photorefractory starlings does not induce immediate testicular growth but may initiate a process which eventually terminates photorefractoriness in a way similar to that caused by return to short daylengths.     

Time Course of Sensitivity of the Photoinducible Phase to Light in the Redheaded Bunting, Emberiza bruniceps

This study investigated the differential sensitivity of the photoinducible phase (Φi) to light in the redheaded bunting (Emberiza bruniceps). Using a skeleton paradigm, we assessed the rate and magnitude of testicular response as a function of the duration of an inducing light pulse and of the time of its introduction in Φi. For a period of 7 weeks, birds received at an intensity of ~100 lux, the same (6 h) entraining light stimulus with a varied inducing light pulse: 1, 2, 4 or 6 h beginning at zeitgeber time (zt) 11 (6L:5D:1L:12D, 6L:5D:2L:11D, 6L:5D:4L:9D or 6L:5D:6L:7D), or 1h pulses at zt 12 (6L:6D:1L:11D) or zt 16 (6L:10D:1L:7D). The testes were stimulated in all LD alternations, but duration- and time-dependent effects of the light pulse on the rate and magnitude of the testicular response were clearly evident. Illumination of a larger portion of Φi seems to result in higher rates of gonadal growth but there is a duration limit above which there will be no further increase of testicular response.     

Temperature alters the photoperiodically controlled phenologies linked with migration and reproduction in a night-migratory songbird

We investigated the effects of temperature on photoperiodic induction of the phenologies linked with migration (body fattening and premigratory night-time restlessness, Zugunruhe) and reproduction (testicular maturation) in the migratory blackheaded bunting. Birds were exposed for four weeks to near-threshold photoperiods required to induce testicular growth (11.5 L:12.5 D and 12 L:12 D) or for 18 weeks to a long photoperiod (13 L:11 D) at 22°C or 27°C (low) and 35°C or 40°C (high) temperatures. A significant body fattening and half-maximal testicular growth occurred in birds under the 12 L, but not under the 11.5 L photoperiod. Further, one of six birds in both temperature groups on 11.5 L, and four and two of six birds, respectively, in low- and high-temperature groups on 12 L showed the Zugunruhe. Buntings on 13 L in both temperature groups showed complete growth-regression cycles in body fattening, Zugunruhe and testis maturation. In birds on 13 L, high temperature attenuated activity levels, delayed onset of Zugunruhe by about 12 days, reduced body fattening and slowed testicular maturation. The effect of temperature seems to be on the rate of photoperiodic induction rather than on the critical day length. It is suggested that a change in temperature could alter the timing of the development of phenologies linked with seasonal migration and reproduction in migratory songbirds.     

Differential responses of the photoperiodic clock in two passerine birds possessing a strongly self-sustained circadian system

To investigate whether the photoperiodic clocks of species possessing strongly self-sustaining circadian clocks share identical features, we compared the full response cycle (initiation and termination of the response) in body mass and testes of the non-migratory house sparrow (Passer domesticus) with that of the migratory redheaded bunting (Emberiza bruniceps) under Nanda-Hamner experiments. Birds were exposed to a 36 h day (L:D=6:30 h), controls exposed to a 24 h day (L:D=6:18 h), for a period of 31 weeks. By week 18 of L:D=6:18 h, there was a small increase in body mass among sparrows, but not among buntings, and the testes of bunting did not grow, while those of sparrow grew slightly. The response to L:D=6:30 h is of particular interest. There was a rapid gain and subsequent loss in the body mass of bunting, but not of sparrows. Further, both species underwent a testicular cycle as if they were exposed to long days, but the response of sparrows was slower and hence delayed the attainment of peak testicular size. Such a differential response to exotic light cycles between these two photosensitive species, despite their similar circadian oscillatory properties (strong self-sustainment), could suggest a species-specific adaptation of the endogenous clock involved in photoperiodic regulation of avian seasonality.     

Differential responses of the photoperiodic clock in two passerine birds possessing a strongly self‐sustained circadian system

To investigate whether the photoperiodic clocks of species possessing strongly self‐sustaining circadian clocks share identical features, we compared the full response cycle (initiation and termination of the response) in body mass and testes of the non‐migratory house sparrow (Passer domesticus) with that of the migratory redheaded bunting (Emberiza bruniceps) under Nanda‐Hamner experiments. Birds were exposed to a 36 h day (L∶D=6∶30 h), controls exposed to a 24 h day (L∶D=6∶18 h), for a period of 31 weeks. By week 18 of L∶D=6∶18 h, there was a small increase in body mass among sparrows, but not among buntings, and the testes of bunting did not grow, while those of sparrow grew slightly. The response to L∶D=6∶30 h is of particular interest. There was a rapid gain and subsequent loss in the body mass of bunting, but not of sparrows. Further, both species underwent a testicular cycle as if they were exposed to long days, but the response of sparrows was slower and hence delayed the attainment of peak testicular size. Such a differential response to exotic light cycles between these two photosensitive species, despite their similar circadian oscillatory properties (strong self‐sustainment), could suggest a species‐specific adaptation of the endogenous clock involved in photoperiodic regulation of avian seasonality.     

Melatonin blocks inhibitory effects of prolactin on photoperiodic induction of gain in body mass,testicular growth and feather regeneration in the migratory male redheaded bunting (<Emphasis Type="Italic">Emberiza bruniceps</Emphasis>)

Little is known about how hormones interact in the photoperiodic induction of seasonal responses in birds. In this study, two experiments determined if the treatment with melatonin altered inhibitory effects of prolactin on photoperiodic induction of seasonal responses in the Palearctic-Indian migratory male redheaded bunting Emberiza bruniceps. Each experiment employed three groups (N = 6–7 each) of photosensitive birds that were held under 8 hours light: 16 hours darkness (8L:16D) since early March. In the experiment 1, beginning in mid June 2001, birds were exposed to natural day lengths (NDL) at 27 degree North (day length = ca.13.8 h, sunrise to sunset) for 23 days. In the experiment 2, beginning in early April 2002, birds were exposed to 14L:10D for 22 days. Beginning on day 4 of NDL or day 1 of 14L:10D, they received 10 (experiment 1) or 13 (experiment 2) daily injections of both melatonin and prolactin (group 1) or prolactin alone (group 2) at a dose of 20 microgram per bird per day in 200 microliter of vehicle. Controls (group 3) received similar volume of vehicle. Thereafter, birds were left uninjected for the next 10 (experiment 1) or 9 days (experiment 2). All injections except those of melatonin were made at the zeitgeber time 10 (ZT 0 = time of sunrise, experiment 1; time of lights on, experiment 2); melatonin was injected at ZT 9.5 and thus 0.5 h before prolactin. Observations were recorded on changes in body mass, testicular growth and feather regeneration.     

Inhibition of cyclic 3′5′ monophosphate synthesis in rat testis by L-triiodothyronine

Summary Cytosolic adenylate cyclase activity from rat seminiferous tubules is inhibited by L-triiodothyronine (L-T₃). In a typical dose-response curve, using Mn-ATP as substrate, no effect is observed at 10⁻¹⁰ M L-T₃; about 15 to 25% inhibition is found in the range between 10⁻⁹ and 10⁻⁶ M L-T₃ and finally a sharp enzyme inhibition is evident at increasing hormone concentrations from 10⁻⁶ to 10⁻⁴ M. Incubation of decapsulated testes with L-T₃ leads to a decrease of intracellular cyclic AMP levels. Dose-response relationships for such effect are similar to those found for adenylate cyclase activity. In this case a clear response is observed at 10⁻⁸ M L-T₃.     

Regulation of seasonality in the migratory male blackheaded bunting (Emberiza melanocephala)

The present study was carried out on a Palearctic-Indian migratory species, the blackheaded bunting (Emberiza melanocephala), to understand the importance of photoperiodism and circannual rhythms in determining seasonality in changes in body mass and testis size in birds. An initial experiment determined the effects of duration and intensity of light on photoperiodic induction. The birds were exposed to different photoperiods (hours of light:hours of darkness; 11.5L:12.5D, 12L:12D, 12.5L:11.5D and 13L:11D) at the same (approximately 450 lux) light intensity, and to 13L:11D at different light intensities (50-, 100-, 400-, 800- and 1000-lux). The induction and subsequent regression of photoperiodic responses were dependent upon duration and intensity of the light period until these reached threshold. A second experiment investigated if an endogenous seasonal rhythm underlies photoperiodism in buntings. Birds maintained since February on a 8L: 16D photoperiod (a non-inductive short day length invariably used to ensure photosensitivity in photoperiodic species) were subjected periodically to 16L:8D (a long day length), one group every month from mid-March to mid-August. The magnitude of long day response in body mass and testes decreased as the duration of the short days progressed, but testicular response was restored in birds that were exposed to long days in July and August. The birds exposed simultaneously to short, long, and natural day lengths for 32 weeks underwent an induction-regression cycle under long days and natural day lengths, but not under short days in which a decrease in body mass occurred after about 20 weeks. The last experiment examined the importance of latitudinal migration on photoperiodism, by comparing the response to long days of three groups which included birds from populations those were held in the outdoor aviary for 1 or 2 years at 27 degrees N and those immediately arrived from their breeding grounds (approximately 40 degrees N). There was no difference in the photoperiodic induction among the three groups, indicating that neither experience to changing photoperiods during a migratory journey, nor to long photoperiods at breeding grounds, were critical for a subsequent response (initiation-termination-reinitiation) cycle. Taken together, these findings suggest that (1) the blackheaded bunting has its own endogenous timing program, which is regulated by the photoperiod, and (2) the photoperiodic programs of bunting are flexible enough to accommodate variations in the amplitude of environmental cycles. Thus, it appears that photoperiodism has evolved independently of the evolution of migration in this species.     

Photoperiodic regulation of seasonal responses in Indian weaver bird (Ploceus philippinus)

Photoperiod (=day length) is the vital factor for the regulation of behavioral and physiological activities in many avian species. This study investigated the seasonal cycles of testicular growth and secondary sexual characteristics of Indian weaver bird under natural day length (NDL) and the effects of duration and intensity of light on photoperiodic induction. In the first experiment, groups of birds (n = 7 each) were exposed to under NDL in April 2008 and May 2009 for 8 and 12 months, respectively. In second and third experiment, birds (n = 6 each group) were exposed to different photoperiods (11.5L:12.5D, 12L:12D, 13L:11D, and 15L:9D) at the same (500 lux) light intensity, and to 13L:11D at different light intensities (10-, 50-, 500-, and 800-lux). Observations on testis size, molt, and plumage score were recorded 2-week (molt and plumage) or at 4-week intervals (testes). Both the NDL groups showed similar seasonal cycles of testicular growth-regression and secondary sexual characteristics. Second and third experiments suggest that the photoperiodic induction was depending upon duration and intensity of the light. Birds showed testicular growth-regression cycle followed by molt and plumage color change only under 13L:11D and 15L:9D and only 500- and 800-lux under 13L:11D photoperiod but not under 11.5L:12.5D and 12L:12D and 10- and 50-lux light intensities. Pre- and post-nuptial molting on body feathers were progressed with gonadal stimulation–maturation and regression cycle under 13L:11D and 15L:9D. Results under different light–dark cycles suggest that day length of about 12 h or more and above the threshold level of light intensity are essential for the induction of photoperiodic responses.     

Seasonal modulation of diurnal food consumption in Indian songbirds

Seasonal changes in daily food consumption have a direct bearing with energy requirement of bird that is in turn associated with life history stage of birds. We compared seasonal changes in daily food intake in adult male migratory redheaded bunting (Emberiza bruniceps) that over winters in Indian subcontinent with those in non-migratory blackheaded munia to reiterate the same. We also compared daily food eating pattern (DFEP) in wintering blackheaded and redheaded buntings, closely related Emberizidae finches to establish circadian nature of feeding behavior and how it varied at species level. The birds were held under short days (8L:16D; 8 h of light and 16 h of darkness) and two hourly food consumption was measured to profile their DFEP. Further, we extended the study to establish how the circadian pattern of food consumption varied depending on birds’ physiological state and effect of photoperiod in adult male redheaded buntings. Redheaded buntings DFEP and locomotor activity were compared in pre-migratory months of February (spring) and September (autumn). The results suggest that September (photorefractory) birds exhibit clear bimodality in their feeding behavior as compared to (photosensitive) birds in February. Another experiment compared bird’s DFEP held under short (8L:16D) and long (16L:8D) days for 5 weeks and suggested that under long days, prolonged hours of photophase render adaptive advantage to birds for positive energy budgeting. The present study clearly establishes the circadian nature of feeding behavior and that it modulates over seasons. The bimodal i.e. morning and evening peaks of food consumption suggest morning–evening food entrainable oscillators, however this needs to be investigated with mechanistic approach in future studies.     

Daily light regulates seasonal responses in the migratory male redheaded bunting (Emberiza bruniceps)

This study analyzed the role of day length in regulation of seasonal body fattening and testicular growth in a latitudinal Palaearctic-Indian migrant, the redheaded bunting (Emberiza bruniceps). When exposed to increasing photoperiods (hours of light: hours of darkness; 11.5L:12.5D, 12L:12D, 12.5L:11.5D, 13L:11D, 14L:10D, and 18L:6D) for 9-12 weeks, buntings responded in a photoperiod-dependent manner and underwent growth and regression cycle under photoperiods of > or =12 hr per day. Also, the response to a long photoperiod of birds that were held under natural photoperiods at 27 degrees N for 2 years was similar to those who arrived the same year from their breeding grounds ( approximately 40 degrees N), suggesting that the experience of higher amplitude day-night (light-dark, LD) cycles during migratory and breeding seasons were not critical for the subsequent response (initiation-termination-reinitiation) cycle. Another experiment examined entrainment of the circadian photoperiodic rhythm in buntings by subjecting them to T=24+/-2 hr LD-cycles with 8 hr photophase and to T=22 and 24 hr with 11 hr photophase. The results showed a reduction in critical day length under T=22 hr LD-cycle. In the last experiment, we constructed an action spectrum for photoperiodic induction by exposing birds for 4.5 weeks to 13L:11D of white (control), blue (450 nm), or red (640 nm) light at irradiances ranging from 0.028 to 1.4 W m(-2). The threshold light irradiance for photoinduction was about 10-fold higher for blue light, than for red and white lights. These results conclude that the daily light of the environment regulates the endogenous program that times seasonal responses in body fattening and testicular cycles of the redheaded bunting.     

Time Course of Sensitivity of the Photoinducible Phase to Light in the Redheaded Bunting,Emberiza bruniceps

This study investigated the differential sensitivity of the photoinducible phase (Φi) to light in the redheaded bunting (Emberiza bruniceps). Using a skeleton paradigm, we assessed the rate and magnitude of testicular response as a function of the duration of an inducing light pulse and of the time of its introduction in Φi. For a period of 7 weeks, birds received at an intensity of ~100 lux, the same (6 h) entraining light stimulus with a varied inducing light pulse: 1, 2, 4 or 6 h beginning at zeitgeber time (zt) 11 (6L:5D:1L:12D, 6L:5D:2L:11D, 6L:5D:4L:9D or 6L:5D:6L:7D), or 1h pulses at zt 12 (6L:6D:1L:11D) or zt 16 (6L:10D:1L:7D). The testes were stimulated in all LD alternations, but duration- and time-dependent effects of the light pulse on the rate and magnitude of the testicular response were clearly evident. Illumination of a larger portion of Φi seems to result in higher rates of gonadal growth but there is a duration limit above which there will be no further increase of testicular response.     

The influence of light wavelength on reproductive photorefractoriness in migratory blackheaded bunting (Emberiza melanocephala).

There are two effects of long day length on reproductive responses in birds, one is the photoinduction of gonadal growth and maturation and the other is the induction of gonadal regression and photorefractoriness. Although it is likely that the same photoreceptors are involved in the photoinduction of gonadal growth and the onset and maintenance of photorefractoriness. and so the influence of wavelength should be similar, this has not been investigated. Therefore, we investigated the influence of light wavelength on reproductive photorefractoriness in the migratory male blackheaded bunting held under long photoperiods. In mid May, when photoperiod was approximately 14L:10D (14 hours light:10 hours darkness), eight groups of sexually mature birds were moved indoors on an artificial photoperiod of 14L:10D (L - 450 lux. D - 0 lux). Then after 3 weeks, for six groups, a 4-h light period in the morning (zt 0-4; zt 0 [zeitgeber time 0] refers to the beginning of lights-on period) or in the evening (zt 10-14) was substituted with green (428 nm), red (654 nm) or white light at 16 +/- 2 lux intensity. Of the remaining two groups, one was maintained on 14L: 10D and the other transferred to 10L:14D: these served as controls. At the end of 4 weeks, all birds were found to have undergone testicular regression, irrespective of LD cycle they were exposed to. When these gonadally regressed birds were subjected to 16L:8D for another 4 weeks, to test their responsiveness to the stimulatory effects of long day lengths, only those exposed to 10L:14D and 14L:10D with a 4-h green light period showed testicular regrowth. On the other hand, birds exposed to 14L:10D with a 4-h white or red light period remained fully regressed, similar to 14L:10D controls. Except for some individual difference, there was no difference in response between the groups that received a 4-h light period in the morning and that received it in the evening. These results suggest that the wavelengths of light influence induction of buntings from the photosensitive state into the photorefractory state. Whereas the short light wavelengths facilitated recovery from the photorefractoriness, the long light wavelengths were more effective in maintaining the photorefractoriness.     

Photosensitivity in body mass and testicular activity of brahminy myna, Sturnus pagodarum

This study analyzed photoperiodic sensitivity of brahminy myna (Sturnus pagodarum), which is a seasonally breeding bird species. During regression phase of the reproductive cycle (in early September), groups of myna were exposed to artificial photoperiods that were either close to or longer than those brahminy myna experiences at this time in wild at 29 degrees N. Following a 14-week exposure to such photoperiods (hours of light: hours of darkness; 13L:11D, 12L:12D, 11.5L:12.5D and 11L:13D), the birds were subjected to a longer day length (16L:8D) for another 9 weeks to test whether pre-treatment with varying photoperiods had an effect on subsequent long day photostimulation. There was a progressive increase in body mass under different pre-treatment photoperiods, with a faster increase in 11L:13D and 11.5L:12.5D than in 12L:12D and 13L:11D. When subjected to 16L: 8D, however, all groups showed decline in body mass. By contrast, the testes were not stimulated under 11L:13D and 11.5L:12.5D, and only one individual of the groups under 12L:12D and 13L: 11D showed a small testis recrudescence. All birds except one individual of the 13L:11D group, however, showed testis recrudescence when subjected to 16L:8D. These results suggest that body mass and testes have a different profile of photoperiodic response and appear to have two different threshold photoperiods for dissipating the post-reproductive refractoriness.     

Wavelength Dependency of Light-Induced Effects on Photoperiodic Clock in the Migratory Blackheaded Bunting (Emberiza melanocephala)

The effects of light wavelength on photoperiodic clock were determined in the migratory male blackheaded bunting (Emberiza melanocephala). We constructed an action spectrum for photoperiodic induction (body fattening, gain in body mass, and gonadal recrudescence) by exposing birds for 4.5 weeks to 13 h light per day (L:D = 13:11 h) of white (control), blue (450 nm), or red (640 nm) color at irradiances ranging from 0.028 to 1.4 W m^?2. The threshold light irradiance for photoinduction was about 10-fold higher for blue, compared to red and white light. Phase-dependent effects of light wavelength on the photoperiodic clock were further examined in the next two sets of skeleton photoperiods (SKPs). In the first set of SKPs, birds were exposed for four weeks to asymmetrical light periods (L:D:L:D = 6:6:1:11 h) at 0.25 ± 0.01 W m^?2; two light periods applied were of the same (450 nm: blue:blue, B:B; 640 nm, red:red, R:R) or different (blue:red, B:R or red:blue, R:B) wavelengths, or of white:white (W:W, controls). Photoperiodic induction occurred under R:R and B:R, but not under B:B and R:B light conditions; the W:W condition induced an intermediate response. The second set of SKPs used symmetrical light periods (L:D:L:D = 1:11:1:11 h), and measured effects also on the activity rhythm. Birds were first exposed to one of the four SKPs (R:R, B:B, R:B, or B:R) for three weeks, subsequently were released into dim constant light (LL_dim; ?0.01 W m^?2, the night light used in an L:D cycle) for two weeks, and then were returned to respective SKPs for another three weeks. Activity was greater in the R:R compared to B:B, and in B:R compared to R:B light condition. Zugunruhe (intense nighttime activity, indicating migratory restlessness in a caged situation) developed under the R:R and B:R, but not the B:B and R:B, light condition. Under LL_dim, all birds free-ran with a period >24 h, the Zugunruhe had a circadian period longer than the daytime activity, and the re-entrainment to SKPs was influenced by the position of light periods relative to circadian phase of the activity rhythm. Photoperiodic induction at the end of 8 weeks was found in the R:R and B:R, but not in B:B, light conditions; in the R:B condition only one bird had initiated testes. Taken together, these results suggest that in the blackheaded bunting, the circadian photoperiodic clock is differentially responsive to light wavelengths; this responsiveness is phase-dependent, and the development of Zugunruhe reflects a true circadian function. Wavelength-dependent response of the photoperiodic clock could be part of an adaptive strategy in evolution of the seasonality in reproduction and migration among photoperiodic species under wild conditions.