Relationships between environmental changes, maturity, growth rate and plasma insulin-like growth factor-I (IGF-I) in female rainbow trout

Size reflecting growth rate, energy balance or nutritional status is regarded as an important determinant of the ability of trout to undergo puberty. The relationship of a change in photoperiod, either natural (SNP) or advancing (ADV), with growth, IGF-I and reproduction was investigated in virgin female rainbow trout. Under SNP 63% of the population attained maturity while only 29% spawned 6 months in advance in the ADV regime. Under SNP both size and growth rate in late spring-early summer appeared to determine whether an individual may initiate reproduction while condition factor appeared to be a better predictor in the ADV regime. A complete seasonal relationship between plasma IGF-I, daylength and temperature was demonstrated under natural conditions, and provides direct evidence for the relationship between reproduction and IGF-I. Conversely, trout maintained under ADV exhibited a significantly different plasma IGF-I profile relative to those under a natural photoperiod. Furthermore, IGF-I levels accurately reflected growth rate prior to elevations in sex steroids, suggesting that IGF-I may provide an endocrine signal between the somatotropic and reproductive axes that growth rate and/or size is sufficient to initiate gonad development. In addition, maturing individuals under SNP typically expressed higher circulating IGF-I levels than those that remained immature and may reflect a greater opportunity for IGF-I to act on the pituitary to stimulate gonadotropin production. We observed elevated levels in maturing fish for 3 months under SNP compared to only 1 month under ADV were observed. This may reflect a reduction in the window of opportunity to initiate reproduction under advancing photoperiods and hence explain the reduction in fish successfully recruited.     

Indoleamines and 5-methoxyindoles in trout pineal organ in vivo: daily changes and influence of photoperiod

This study describes the diel rhythms in several indoleamines, melatonin, and related 5-methoxyindoles in the pineal organ of rainbow trout in vivo. In addition, the effect of different photoperiod conditions was evaluated. Melatonin levels displayed clear daily rhythms in the pineal organ of rainbow trout kept experimentally under long (LD 16:08), neutral (LD 12:12), and short (LD 08:16) photoperiods. Duration of melatonin signal was dependent on the night length of prevailing photoperiod, while peak amplitude was higher when lengthening the photoperiod. Significant daily rhythms in 5-HT content, the precursor of melatonin synthesis, were found in neutral and short photoperiod with increases of the amine content just after the light-dark interphase and decreases in the middle of the night, which were more important under short photoperiod. In contrast, no significant 24-h cyclic variation was found in pineal 5-HT content under long photoperiod. Daily profiles in the content of the main 5-HT oxidative metabolite, the 5-hydroxyindoleacetic acid (5-HIAA), outlined those of the amine precursor. The chronograms of both aminergic compounds contrast with those of 5-hydroxytryptophan content, which displayed a net tendency to increase at night. This study also provides evidence for the existence of daily cyclic changes in the content of 5-methoxytryptamine (5-MT), 5-methoxyindoleacetic acid (5-MIAA), and 5-methoxytryptophol (5-MTOL) in trout pineal organ, which were also dependent on photoperiod. The 24-h profiles in 5-MT content correlated well with those of 5-HT, showing a peak at the first hour of darkness in all photoperiodic conditions, and a decay at midnight only in both neutral and long photoperiods. Similarly, the content of 5-MTOL also displayed high values during the day-night transition in trout kept under neutral and long photoperiods, followed by a slow decay all along the night. Finally, levels of 5-MIAA increased in all photoperiods when lights were turned off, being this nocturnal increase maximal in fish kept under LD 16:08. These results suggest that light-dark cycle modulates daily rhythms in pineal indoles and non-melatonin 5-methoxyindoles by acting mainly through the melatonin synthesis activity, which limits the availability of 5-HT for the oxidative and direct methylation pathways. In addition, it seems that a nocturnally increased synthesis of 5-HT might be a requirement for the optimal formation of melatonin and other 5-methoxyindoles in the pineal organ when trout remain under short photoperiods.     

The brain-pituitary-gonadal axis of female rainbow trout Oncorhynchus mykiss: effects of photoperiod manipulation1.

Two groups of post-spawned female rainbow trout were exposed to two different photoperiods, an ambient photoperiod (56 degrees N) and a combination of long and short photoperiods (a constant 18L:6D from February 1 until May 10, then a constant 6L:18D), which acted to advance maturation and spawning. The stimulatory long-short photoperiod advanced spawning by 3-4 months and correspondingly advanced peaks in serum levels of 17beta-estradiol, testosterone, calcium (an index of vitellogenin), and GTH II. Earlier events in gonadal recrudescence appeared to be less affected by the photoperiod. The initiation of exogenous vitellogenesis coincided with high levels of both pituitary salmon gonadotropin-releasing hormone (sGnRH) content and serum follicle-stimulating hormone (FSH, GTH I) levels. High levels of serum FSH were associated with rapid gonadal growth in the fish exposed to the stimulatory long-short photoperiod. In contrast, the fish exposed to the ambient photoperiod showed gonadal steroid production, formation of vitellogenin, and secondary oocyte growth without any detectable increase in serum FSH levels. The possible roles and interactions of sGnRH, gonadotropins, and steroids with respect to normal and artificially stimulated ovarian maturation are discussed.     

Plasma prolactin and luteinizing hormone levels of pinealectomized and enucleated turkey hens

Plasma levels of LH and prolactin were determined in pinealectomized (PX) and/or bilateral ocular enucleated (EX) adult turkey hens maintained in either constant (24LL) or diurnal (16L:8D) lighting. Blood samples were obtained at the following times over a 24-hr period: onset, middle, and end of light period, middle and end of dark period. In experiment 1, the effects of PX alone were evaluated in photorefractory hens exposed to two different photoperiods (24LL and 16L:8D). In experiment 2, the effects of PX and EX were determined in egg-laying and photorefractory hens exposed to a 16L:8D daily photoperiod. Neither PX nor EX significantly altered LH or prolactin levels of egg laying or photorefractory hens at any of the sampling times of the day in either experiment. In experiment 1, LH levels were very similar in both 16L:8D and 24LL but prolactin levels were significantly lower in 24LL than 16L:8D. It was concluded that the pineal gland does not play an essential role in maintaining circulating LH and prolactin levels in constant light or a photoperiod of 16L:8D.     

Old rats are more sensitive to photoperiodic changes. A study on pineal melatonin

After having previously demonstrated that beta-adrenergic stimulation of melatonin under a standard light:dark (LD) cycle regimen of 12:12 is more effective in young than in old pineal glands, we have now studied how the daylength change LD 18:6 affects pineal melatonin secretion and its regulation by the beta-adrenergic system. Young (10 weeks) and old (22 months) male Wistar rats were synchronized with either a standard LD 12:12 for 4 weeks, or acclimatized under the same LD conditions for 4 weeks, then subjected to a long LD 18:6 photoperiod for 1 week. The rats were sacrificed at three time samplings: 0, 4, and 7h after dark onset (HADO) for LD 12:12 or 0, 2, and 3.5 HADO for LD 18:6. Pineal glands were collected and perifused for 480 min. Isoproterenol (10(-4)M) was infused for 20 min, 4h 10 min after the beginning of the perifusion. Basal levels of melatonin production in the young rats displayed a 1.5-2.5-fold increase compared to those in the old rats. Interestingly, mean basal melatonin levels in old rats under standard LD 12:12 conditions were significantly higher (P<0.05) than mean levels at the same relative dark phase intervals under LD 18:6 conditions. Isoproterenol stimulated melatonin production in both young and old rat pineal glands, regardless of time sampling or photoperiodic conditions. The magnitude of the response to 10(-4)M isoproterenol infusion in old pineals was approximately half that found in young glands (P<0.001), and tended to be higher under LD 12:12, in both young and old rat pineal glands, although no significant difference was found in melatonin response between the two photoperiods (P>0.05). This study shows that basal pineal melatonin levels in old rats are more sensitive to photoperiod changes than in young rats. These results also demonstrate that isoproterenol can stimulate both young and old rat pineal glands irrespective of time or photoperiod and confirm previous findings, showing that the melatonin response to isoproterenol is age-dependent and that pineal gland response to isoproterenol is not photoperiod-dependent, at least under our experimental conditions.     

Adjustment of pineal melatonin and N-acetyltransferase rhythms to change from long to short photoperiod in the Djungarian hamster Phodopus sungorus

In the Djungarian hamster Phodopus sungorus, the daily temporal pattern of synthesis and release of pineal hormone melatonin, mainly the length of the period of elevated melatonin levels, may be involved in transferring the information on day length to the neuroendocrine-gonadal axis. The present study investigated the time course of adjustment of the rhythm in melatonin production and concentration to the change from long to short photoperiods. Adult female Djungarian hamsters, maintained on a regime of 16 h of light and 8 h of darkness per day (LD 16:8) were transferred to the LD regime 8:16 and the daily rhythms in the pineal melatonin concentration and in the pineal N-acetyltransferase activity, as an indicator of melatonin formation, were studied at various intervals following the transfer. Under LD 16:8, the nocturnal melatonin concentration was elevated for 4.8 h. After 3 days on LD 8:16, no extension of the period of high melatonin levels occurred. 2, 4 and 6 weeks after the transfer to LD 8:16, the period of elevated melatonin levels lasted for 8.1, 9.3 and 11.5 h, respectively. Extension of the melatonin pattern proceeded first predominantly into the morning hours. Only after this extension was completed, a considerable extension into the evening hours began. Extension of the N-acetyltransferase rhythm on short photoperiods proceeded in the same way as that of the melatonin rhythm. The data show that while a change in the photoperiod might be seen by hamsters within 2 weeks after the transfer to LD 8:16, the full shortening of the photoperiod might be recognized only within 6 weeks or later.(ABSTRACT TRUNCATED AT 250 WORDS)     

The circadian nature of melatonin secretion in Japanese quail (Coturnix coturnix japonica)

Abstract: Plasma melatonin concentrations were measured in Japanese quail held under different photoperiods and constant darkness (<1 lux). When subjected to LD6:18 (6 hr light: 18 hr darkness), levels rose ～2 hr after lights-off, attained a peak level 8 hr after lights off, and subsequently declined to low daytime levels before the next lights-on signal. This generated a distinct daily rhythm in melatonin secretion with a duration of ～13 h. On exposing quail to a range of photoperiods, containing 6, 9, 11, 12, 13, 15, 18, or 20 hr of light per day, the onset of melatonin secretion remained essentially similar with the rise occurring soon after lights-off. However, the offset of melatonin secretion was suppressed by the light of the next day and thus a much truncated rhythm was produced under long (> 12 hr) photoperiods. Importantly, between night lengths of 4 to 18 hr (i.e., LD 20:4 to LD 6:18) a linear relationship existed between the duration of night-length and secretion of melatonin with the duration increasing by about 0.8 hr for each additional hour of darkness. If quail were released into darkness following a short (LD 6:18) or long (LD 20:4) day schedule, the rhythm persisted for at least two cycles with peaks occurring at about 24 hr intervals. In those quail coming into darkness from long days (LD 20:4), the rhythm of melatonin secretion decompressed rapidly on both sides of the peak, indicating that both the onset and offset of melatonin secretion were suppressed under long days. The endogenous nature of melatonin secretion was tested further by exposing birds to LD 6:30 for 4 cycles and then releasing into darkness. The rhythm in melatonin secretion persisted for at least three cycles before beginning to damp-out. The circadian nature of the rhythm in melatonin secretion was also examined by subjecting quail to T-cycles and then releasing into darkness. Both under the T-cycles and darkness following T-cycle treatments, the phase of the melatonin rhythm was advanced by > 3 hr under T = 27 hr cycles (LD 3:24) compared with T = 24 hr cycles (LD 3:21). This property is consistent with the melatonin oscillator being a circadian rhythm.     

Growth hormone and insulin-like growth factor-I act together and independently when regulating growth in vertebral and muscle tissue of atlantic salmon postsmolts

Aiming to elucidate the role of GH and IGF-I with regard to vertebral and white muscle growth, gene expression of the GH and IGF-I receptors (ghr and igf-Ir, respectively) and local IGF-I (igf-I) were analyzed during spring growth (January-June) in Atlantic salmon postsmolts. One group of fish was reared under natural light (NL), while one group was reared under continuous light (LL). Growth rate of fork length was higher in the LL group for a short period after onset of continuous light (LL: 0.50+/-0.02 mm day(-1), NL: 0.43+/-0.01 mm day(-1)) and for a longer period at the end of the experiment in June (LL: 1.18+/-0.06 mm day(-1), NL: 0.75+/-0.02 mm day(-1)). Likewise, growth rate in length of vertebra No. 40 in the LL group was higher than in the NL group the first period after onset of light (LL: 0.015+/-0.002 mm day(-1), NL: 0.008+/-0.001 mm day(-1)). Plasma GH levels peaked in late February and were higher in the LL group than in the NL group (LL: 7.27+/-0.61 ng ml(-1), NL: 2.60+/-0.50 ng ml(-1)), whereas plasma IGF-I levels peaked in early February and were unaffected by photoperiod. ghr expression was upregulated in late February in liver (12-fold), white muscle (6-fold) and vertebral tissue (3-fold) and higher in the LL group than in the NL group (2-fold) in vertebral tissue in late March. White muscle expression of igf-I and igf-Ir decreased from initial levels throughout the experiment. Hepatic gene expression of igf-I doubled in both groups in late February, followed by a 4-fold upregulation in June in the LL group only. Vertebral tissue expression of igf-I (4-fold) and igf-Ir (6-fold) increased in May and were unaffected by photoperiod. One exception was a smaller upregulation of igf-I (2-fold) in the LL group in early February. In conclusion, GH appears to have an initial role in stimulating vertebral growth, while IGF-I seems to stimulate growth during late spring. It is suggested that local IGF-I acts as a paracrine agent, evaluated from the concurrent upregulation of igf-I and igf-Ir. The upregulation of ghr in white muscle tissue, concurrent with a downregulation of muscle igf-I and igf-Ir, indicate that GH stimulated growth or metabolism independent of IGF-I.     

Melatonin Rhythms in Quail: Regulation by Photoperiod and Circadian Pacemakers

The profile of melatonin in the eyes, pineal, and blood of Japanese quail was assessed in birds held under LD 16:8 and LD 6: 18 photoperiods. Melatonin levels in all three tissues showed a robust daily rhythm with higher levels occurring at night. The amplitude of the rhythm was depressed and its duration lengthened on LD 6: 18 relative to LD 16:8. The blood melatonin rhythm precisely reflected the rhythms shown by the pineal and eyes, supporting the idea that the blood rhythm is a result of melatonin secretion by both the eyes and pineal.
The ocular melatonin rhythm continued after sectioning of the optic nerve, was reentrainable to a shift in the phase of the LD cycle, and persisted for at least 2 days in constant darkness. It was concluded that either (1) an intraocular circadian clock drives the ocular melatonin rhythm, or (2) an extraocular clock drives the ocular melatonin rhythm via a route other than the efferent innervation (which enters the eye via the optic tract).     

Cytosolic androgen receptors in the neuroendocrine tissues of the golden hamster: influence of photoperiod and melatonin treatment

Hamsters exposed for eight weeks to short photoperiod (LD 10:14) or treated with melatonin in the late afternoon under long photoperiod (LD 14:10) had significantly higher number of cytosolic androgen receptors in the pituitaries, hypothalami and harderian glands, as compared to the long photoperiod (LD 14:10) exposed controls. The numerical value of the apparent Kd was two to three times lower in the hypothalami and pituitaries, but not in the harderian glands of the animals from these groups. These results indicate that alterations in receptor numbers and affinity constants may be responsible for the dramatic changes in the sensitivity of the hypothalamo-pituitary axis to the negative feedback actions of the gonadal steroids, observed under inhibitory photoperiods and that this effect could be duplicated by late afternoon melatonin treatment.     

Melatonin mediates photoperiod control of endocrine adaptations and humoral immunity in male Siberian hamsters

Effects of photoperiod are mediated by the pineal gland in male Siberian hamsters. The hypothesis that the pineal hormone melatonin mediates the effects of short days (SD) to blunt select humoral and endocrine functions was tested. In the first study, regressed testes were found in pineal-intact controls transferred from long days (LD) to SDs (16 hr to 8 hr light/day); the rise in antigen-induced serum immunoglobulin (Ig) M was blunted and serum cortisol concentrations elevated compared with long-day controls. These effects of short-day were blocked in pinealectomized males moved from long to SDs, but restored by melatonin treatments. In a second study, males in LD were exposed to constant light (LL) to abolish the nighttime melatonin rhythm. In hamsters in LL, melatonin induced testicular regression as in males in SDs. Large testes were present in vehicle-treated controls in LL and in males that remained in LDs. Antigen-induced increases in serum IgM in vehicle and melatonin treatment males in LL were intermediate between concentrations in long- or short-day controls and not significantly different from each other. However, serum cortisol was again elevated in hamsters in SDs or in LL when treated with melatonin compared with males in LL or LDs. These findings indicate that melatonin treatments mimicked the effects of SDs to regulate adaptive physiologic functions in hamsters lacking the nocturnal melatonin rhythm. Thus, the photoneuroendocrine mechanism regulating reproductive responses to photoperiod also mediates short-day effects on T cell-dependent B-cell antibody production and processes that regulate cortisol in circulation.     

Androgenic function in adult rats: Influence of the pineal gland of the mother and of the offspring

Female rats exposed to long (LD 18:6) or short (LD 6:18) photoperiods from 21 days of age were mated when they reached 55 days of age. On day 2 of gestation animals of each group were either pinealectomized or sham-operated. Lighting regimens were not changed during the course of the study. Male offspring of the four groups of dams were sacrificed on day 70 after birth. Rats that were maintained on long photoperiod had higher testicular testosterone, androstenedione, and dihydrotestosterone content than those raised on a LD 6:18 cycle. Whatever the breeding photoperiod used, maternal pinealectomy induced no modification of reproductive function. Among rats kept in short photoperiod, neonatal pinealectomy (on day 5 after birth) resulted in an enhanced testicular androgen content without any modification of plasma androgen concentration. These results indicate that (1) the previously reported effect of the mother's pineal on pubertal rat testicular function is not present in adulthood and (2) the pineal of the offspring is required to maintain normal testicular androgen content in the adult rat but exerts no influence on circulating androgens.     

The effect of photoperiod on diel rhythms in serum melatonin, cortisol, glucose, and electrolytes in the common dentex, Dentex dentex

Diel rhythms in serum concentrations of melatonin, cortisol, glucose, sodium, chloride, and potassium were studied in the common dentex, Dentex dentex, under different photoperiods (DD, 8L:16D, 12L:12D, 16L:8D). Photoperiod affected both the diel rhythms and the absolute values of the estimated blood components. Regardless of the photoperiod, melatonin titers were elevated during the scotophase (384.3 +/- 13.9 pg/ml) compared with a mean baseline level of 54.4 +/- 2.7 pg/ml during the photophase. Serum melatonin concentrations reflected the prevailing photoperiod and constantly elevated melatonin levels with no diel rhythmicity were evident in fish held in the DD protocol. A circadian-like pattern in serum cortisol was observed in fish that were kept at the DD and 8L:16D protocols with cortisol peak at 18:00 h in the night. Fish exposed to the 16L:8D regime showed highest cortisol levels at 10:00 h, while no rhythmicity was evident under the 12L:12D protocol. A phase shift of 4 h between the peaks of cortisol and glucose was evident in fish exposed to the DD, 8L:16D, and 12L:12D regimes. Diel patterns of changes in serum Na+ and Cl- were observed only in the fish held in the DD protocol. Serum K+ values were lowest during the first part of the scotophase under all regimes, except the 16L:8D where no diel rhythmicity was detected. During the photophase, cortisol was positively correlated with glucose, Na+, and Cl- and negatively with K+. During the scotophase, melatonin was positively correlated with glucose and electrolytes. Results indicated that cortisol may be responsible for the observed rhythmicity of glucose and that melatonin may play a role in glucose and ion regulation in common dentex.     

Melatonin and maturation pace in female three-spined stickleback, Gasterosteus aculeatus

In many animals sexual maturation is controlled by the photoperiod. In mammals, the photoperiodic message is mediated via melatonin, but it is unclear whether this also applies to fishes. Administration of melatonin via the water on a schedule aimed at mimicking a short nonstimulatory photoperiod cycle has been found not to inhibit maturation in male three-spined sticklebacks, Gasterosteus aculeatus, kept under stimulatory long photoperiods. To study whether melatonin affects maturation pace, adult female sticklebacks kept under stimulatory photoperiodic regimes of Light:Dark (LD) 24:0 h (Exp. 1) or LD 16:8 h (Exp. 2) and 18 degrees were treated with melatonin (0, 20, or 80 microg/L water) via the water for 16 h per day. In addition, females were also kept under a nonstimulatory short photoperiod (LD 8:16). The time at which full maturation was achieved (running roe) was noted and the ovaries of nonovulated fish were studied histologically. Most fish under LD 24:0 and LD 16:8 matured (maturation rates in Exp. 1 and 2, respectively: control, 100 and 86%; low-dose melatonin, 83 and 93%; high-dose melatonin, 90 and 75%), whereas almost all females kept under LD 8:16 remained immature (maturation rates: 0% in Exp. 1 and 3% in Exp. 2). There was no difference in maturation pace, proportion of fish maturing, or relative ovarian weights between controls and melatonin-treated fish kept under LD 16:8. Furthermore, there was no difference in proportion of fish maturing or relative ovarian weights between controls and melatonin-treated fish kept under LD 24:0. However, LD 24:0 controls matured significantly earlier than fish receiving the high melatonin dose. Thus, there was an inhibitory effect of the high melatonin dose on maturation pace under LD 24:0. Nevertheless, this effect was small compared to the inhibitory effect of LD 8:16 treatment, suggesting that at least in this season, the major part of the photoperiodic effects in the stickleback is mediated via mechanisms other than circulating melatonin.     

Influence of light-dark and lunar cycles on the ocular melatonin rhythms in the seagrass rabbitfish, a lunar-synchronized spawner

The aim of this study was to investigate the effects of light-dark (LD) cycles and lunar phases on ocular melatonin rhythms in the seagrass rabbitfish, Siganus canaliculatus, a lunar-synchronized spawner. Under a natural 24-hr LD (12.00:12.00) cycle, ocular melatonin levels were low during daylight hours. The levels significantly elevated to peak during the mid-dark phase at 24.00 hr and then declined sharply in the early morning around 06.00 hr. These rhythms disappeared under either constant light (LL) or constant dark (DD) conditions. Melatonin levels remained low in LL compared with those in DD condition. These results suggest that ocular melatonin rhythms in the seagrass rabbitfish are suppressed in the presence of light. When fish were exposed to natural moon phases, ocular melatonin concentrations were higher around the new moon than both the first quarter and full moon phases. Exposure to experimental new moon conditions caused a significant increase in melatonin levels while those of the fish exposed to experimental full moon conditions were decreased. These results suggest that the seagrass rabbitfish perceives moonlight through the eye and that moonlight directly causes melatonin suppression.     

Photoperiodic control of melanoma growth in hamsters: influence of pinealectomy and melatonin

Pinealectomy (PX) increased MM1 (melanotic melanoma no. 1) hamster melanoma growth in animals held under a 14-h light, 10-h dark (14:10) photoperiod without altering tumor latency. Hamsters maintained under a 6-h light, 18-h dark (6:18) photoperiod exhibited gonadal collapse, a longer tumor latency, and slower tumor growth rate than animals held under 14:10. PX produced a further increase in tumor latency and a decrease in growth in these animals. In contrast, acute morning injection of low doses (50 micrograms/day) of melatonin or delivery by Silastic capsule (35 micrograms/day) implanted at the time of tumor cell inoculation increased MM1 melanoma growth in hamsters held under 14:10 photocycle, without affecting testicular or adrenal function. Treatment of hamsters 11 weeks before tumor cell inoculation with 14 micrograms/day melatonin via Silastic capsule produced a decrease in serum PRL but no change in tumor growth or testicular or adrenal weights in animals held under 14:10. Treatment of hamsters with 17.7 micrograms/day melatonin (Silastic capsule) 11 weeks before tumor cell inoculation increased testes and adrenal weights as well as serum PRL and androgen levels, but significantly decreased tumor growth in hamsters held under a short daily photoperiod. These results suggest that the photoperiod under which hamsters are maintained dictates the growth rate of MM1 tumors and the effect of PX on tumor behavior. When photoperiod significantly alters gonadal and adrenal function, the quantity, time, and duration of melatonin presentation are all important variables in the effect of melatonin on tumor growth.     

Plasma thyroxine and triiodothyronine levels during the development of photorefractoriness in Willow Ptarmigan (Lagopus lagopus lagopus) exposed to different photoperiods

Changes in the concentrations of plasma thyroxine (T4) triiodothyronine (T3), and luteinizing hormone (LH) are measured in photosensitive Willow Ptarmigan (Lagopus lagopus lagopus) transferred from a 6-hr daylength (6L:18D) to 14-hr (14L:10D), 18-hr (18L:6D), or continuous (LL) daylengths. The increase in plasma LH, and the moult from white to pigmented plumage, observed after transfer to the three stimulatory photoperiods, are not associated with any immediate changes in the concentrations of plasma T4 or T3 except for an increase (P < 0.01) in T4 levels in birds transferred to 18L:6D. As indicated by a fall (P < 0.01) in concentrations of plasma LH, the birds exposed to LL and 18L:6D are becoming photorefractory after 8 to 9 weeks of photostimulation. At this time, the concentration of plasma T3 but not of T4 increases significantly (P < 0.001) and remains high for 5 and 9 weeks in birds exposed to LL and 18L:6D, respectively. Thereafter, levels of plasma T3 begin to fall concomitant with an increase in the levels of plasma T4. In birds exposed to 14L:10D, LH levels remain elevated during the 20-week study and minor increases are observed in the levels of plasma T3 and T4. These observations do not suggest a major role for thyroid hormones in the initiation of photorefractoriness in the Willow Ptarmigan.     

Identification of a second insulin-like growth factor in a fish species.

An internal portion of insulin-like growth factor (IGF) amplified from the total cDNA of rainbow trout (Oncorhynchus mykiss) liver by a PCR was used to screen a rainbow trout liver cDNA library, and recombinant clones encoding two distinct IGFs were isolated. On the basis of a 98.7% nucleotide and 98.3% predicted amino acid identity to coho salmon IGF-I, one cDNA sequence was identified as rainbow trout preproinsulin-like growth factor I (rtIGF-I). The second cDNA sequence shared 46.1% and 43.3% identity with rtIGF-I at the nucleotide and predicted amino acid levels, respectively, and was identified as rainbow trout preproinsulin-like growth factor II (rtIGF-II). Predicted amino acid sequence comparisons of rtIGFs with those of human IGFs indicate that rtIGF-I is more similar to human IGF-I than to human IGF-II, and that rtIGF-II is more similar to human IGF-II than to human IGF-I. Southern blot analysis of rainbow trout genomic DNA probed with rtIGF-I and -II cDNA suggests that these two forms of IGF originate from separate genes. The presence of a teleost IGF-II suggests that the divergence of IGFs occurred early in vertebrate evolution.     

Entrainment of the circadian rhythm in the rat pineal N-acetyltransferase activity by melatonin is photoperiod dependent

Entraining effect of melatonin on the circadian rhythm in rat pineal N-acetyltransferase (NAT) activity was studied under various photoperiods. Melatonin administration prior to dark onset for 5 successive days phase-advanced the evening NAT rise under the light:dark (LD) cycle of either LD 10:14 or LD 8:16, but not under LD 12:12. It is assumed that under the latter regime, the end of a light period exhibited a phase-delaying effect on the NAT rise. The light exposure appeared to be a stronger Zeitgeber than melatonin itself. Data show that melatonin applied in the late light period advances the evening NAT rise under a short photoperiod only; under a longer photoperiod, the phase-advancing effect of melatonin may conflict with a phase-delaying effect of the end of a light period, and the effect of light exposure overrides that of melatonin.     

Examination of in vitro melatonin secretion from superfused trout (Salmo gairdneri) pineal organs maintained under diel illumination or continuous darkness

Melatonin secretion was measured from rainbow trout (Salmo gairdneri) pineal organs maintained individually under flow-through whole organ culture (superfusion) conditions. Radioimmunoassay of perfusate fractions collected during controlled photic conditions demonstrated that melatonin secretion in vitro remained basal during the photophase and underwent increases in titer during the scotophase. While amounts of melatonin (mel) secreted were characteristic of individual pineal organs, photophase values ranged between 0.25 and 0.75 ng mel/ml and scotophase values ranged from 6 to 10 ng mel/ml of perfusate. Diel melatonin secretion profiles reflected the illumination regimen, with light associated with low melatonin titer in the perfusate and darkness associated with high titer. Light pulses during a normal scotophase resulted in a depression in melatonin secretion regardless of whether it was administered early or late in the dark period. Pulses of darkness given early or late in a normal photophase resulted in increased melatonin secretion. Superfused trout pineal organs did not display endogenous rhythmicity in melatonin secretion when subjected to prolonged exposure to continuous darkness (DD), whether first exposed to entraining light/dark (LD) cycles prior to DD or exposed to DD at the initiation of superfusion. In both studies, elevated melatonin secretion gradually declined over time. But exposure to a 4:4LD cycle after DD resulted in decreased (with light) and increased (with darkness) melatonin secretion. These results demonstrate that the trout pineal organ can be maintained for extended periods of time in superfusion culture, that the trout pineal organ is very responsive to light or dark for regulating melatonin synthesis, and that an endogenous rhythm in melatonin synthesis when organs were maintained in DD was not detectable.