未产卵雌性黄鳝的性转变

为探讨产卵是否为雌性黄鳝(Monopterus albus Zuiew)性转变的必经过程, 研究分析了实验室内从受精卵或幼苗开始养殖至不同时间段的黄鳝性腺组织学状况, 采用性腺活检技术跟踪了34月龄雌性黄鳝性腺发育变化, 并以免疫组织化学方法探讨了黄鳝不同发育状态性腺中增殖细胞核抗原(PCNA)的分布。在养殖过程中, 实验黄鳝没有出现产卵现象或者繁殖行为。在5月龄黄鳝中, 间性占比13.3%, 雄性占比20.0%; 在12月龄(1龄)黄鳝中, 雄性占比17.6%; 34月龄(3龄)黄鳝中, 间性占比12.8%, 雄性占比8.5%。通过性腺活检技术对36条34月龄雌性黄鳝性腺发育变化进行了为期1个月的跟踪研究, 结果表明, 16.7%的雌性黄鳝发生了性转变, 性腺发育到间性阶段。黄鳝间性早期性腺生殖褶增厚, 部分细胞呈现明显PCNA免疫阳性, 包括间质细胞、精原细胞和初级精母细胞。上述结果提示, 产卵并非雌性黄鳝发生性转变的必经过程; 黄鳝性转变初期, 性腺结构变化包括生殖褶中间质细胞和精原细胞的发生和增殖。     

Bidirectional sex-change behavior and physiological aspects in the Gorgeous goby Lythrypnus pulchellus (Gobiidae)

The Gorgeous goby Lythrypnus pulchellus shows extreme sexual plasticity with the bidirectional sex-change ability socially controlled in adults. Therefore, this study describes how the hierarchical status affects hormone synthesis through newborn hormone waste products in water and tests the influence of body size and social dominance establishment in sex reversal duration and direction. The associated changes in behavior and hormone levels are described under laboratory conditions in male–male and female–female pairs of similar and different body sizes, recording the changes until spawning. The status establishment occurred in a relatively shorter time period in male and female pairs of different sizes (1–3 days) compared to those of similar size (3–5 days), but the earlier one did not significantly affect the overall time of sex change (verified by pair spawning). The changes in gonads, hormones, and papilla occurred in sex-changer individuals, but the first one was observed in behavior. Courtship started at 3–5 days in male pairs and from 2 h to 1 day in female pairs of both groups of different and similar sizes. Hormones did not gradually move in the new sexual phenotype direction during the sex-change time course. Nonetheless, estradiol regulated sex change and 11-ketotestosterone enabled bidirectional sex change and was modulated by agonistic interactions. Cortisol is associated with status and gonadal sex change. In general, similar mechanisms underlie sex change in both directions with a temporal change sequence in phases. These results shed new light on sex-change mechanisms. Further studies should be performed to determine whether these localized changes exist in the steroid hormone synthesis along the brain–pituitary gonad axis during social and bidirectional sex changes in L. pulchellus.     

黄鳝的繁殖生态学研究

以洞庭湖稻区黄鳝为调查研究对象 ,通过野外调查和室内实验系统研究了黄鳝的繁殖生态。对黄鳝的性腺发育周年变化、繁殖行为、自然产卵过程、产卵条件及繁殖洞穴的构造进行了观察和记录 ,产卵室是黄鳝繁殖洞穴特有的构造 ,繁殖洞穴泥土p H平均值为 6 .73± 1.0 12。亲鳝有护卵习性 ,通过对 30尾守洞亲鳝的性腺观察和切片验证 ,守洞鳝多为雄鳝 (占 6 1.3% ) ,少数为兼性偏雄性 (占 38.7% ) ,守洞鳝 1:0 0～ 2 :0 0全在外面活动 ,守洞鳝体长 L (cm)与体重 W (g)回归方程为 :W=1.2 5×10 - 2 L1 .4 2 (r=0 .76 )。人工模拟条件下 ,泡沫组受精卵的平均孵化率 (85 .2 % )极显著地高于对照组 (2 5 .4 % ) (t=8.18,t0 .0 5=2 .4 5 ) ,仔鱼平均成活率 (6 4 .0 % )显著高于对照组 (14 .0 % ) (t=3.73) ,同时对繁殖季节黄鳝为孵卵而所吐的泡沫的作用进行了分析。另外 ,证实了黄鳝产卵的最适放养密度为 2～ 3尾 / m2。为黄鳝的全人工繁殖和半人工繁殖提供了合理化的建议     

Influence of Altered Photoperiods on Serum Melatonin and Its Receptors (MT1 and MT2) in the Brain,Retina, and Ovary in Carp Catla catla

Daily variation in melatonin receptor (MT1 and MT2) density in three specific tissues—brain, retina, and ovary—and its temporal relationship with serum melatonin were evaluated for the first time in a freshwater teleost, the carp Catla catla, under natural as well as altered photoperiods in different reproductive phases of the annual cycle. Cosinor analysis was used to determine rhythmic features of the serum melatonin and receptors (MT1 and MT2) in different tissues. In each photoperiodic group, irrespective of season, the daily minimum serum melatonin level was noted at midday. However, the daily peak value of melatonin varied in relation to both photo-schedules and reproductive phases. Under natural photoperiods (NPs; duration varied with seasons) and short photoperiods (SPs; light [L]:dark [D] 8:16), it occurred in the late dark phase during the preparatory phase, and at midnight in the remaining parts of the annual cycle. On the other hand, in each reproductive phase, compared to corresponding NP carp, the daily melatonin peak under long photoperiods (LPs; L:D 16:8) exhibited a phase delay of ～2–3?h (occurring during the late dark phase). The melatonin levels at each sampling point were highest during the postspawning phase and lowest during the spawning phase, irrespective of the photoperiodic history of the fish. In each tissue, Western blot analysis revealed a band at ～37?kDa and a band at ～36?kDa corresponding to the molecular weights of native MT1 and MT2 receptor proteins, respectively, with the band intensity of MT1 always being higher than that of a 36-kDa protein. The content of both melatonin receptor proteins varied significantly according to the studied tissue (being highest in the retina, intermediate in the brain, and lowest in the ovary), time in the daily cycle (peak at midnight and fall at midday), and reproductive phase in the annual cycle (highest in the spawning phase and lowest in the postspawning phase). Remarkably, no significant effects of altered photoperiod were detected on any rhythm parameters of either MT1 or MT2 in any of the studied tissues. Collectively, the results of the present study suggest a role of photoperiod in determining daily and seasonal profiles of serum melatonin, but not its receptor proteins, on the ovary or on any nongonad tissues in carp. (Author correspondence: dgp_skmaitra@yahoo.co.in)     

Rhythmic melatonin secretion in different teleost species: an in vitro study

The rhythmic production of melatonin is governed by intrapineal oscillators in all fish species so far investigated except the rainbow trout. To determine whether the latter represents an exception among fish, we measured in vitro melatonin secretion in pineal organs of nine wild freshwater and six marine teleost species cultured at constant temperature and under different photic conditions. The results demonstrate that pineal organs of all species maintain a rhythmic secretion of melatonin under light:dark cycles and complete darkness, and strongly suggest that most fish possess endogenous intrapineal oscillators driving the rhythm of melatonin production, with the exception of the rainbow trout.Abbreviations LD light:dark - DD dark:dark - NAT N-acetyltransferase - RIA radioimmunoassay     

Melatonin in the Regulation of Annual Testicular Events in Carp Catla catla: Evidence from the Studies on the Effects of Exogenous Melatonin,Continuous Light,and Continuous Darkness

The physiological significance of melatonin in the regulation of annual testicular events in a major carp Catla catla was evaluated through studies on the effects of graded dose (25, 50, or 100 µg/100 g body wt.) of melatonin exogenously administered for different durations (1, 15, or 30 days) and manipulation of the endogenous melatonin system by exposing the fish to constant darkness (DD) or constant light (LL) for 30 days. An identical experimental schedule was followed during the preparatory (February–March), pre‐spawning (April–May), spawning (July–August), and post‐spawning (September–October) phases of the annual cycle. Irrespective of the reproductive status of the carp, LL suppressed while DD increased the mid‐day and mid‐night values of melatonin compared to respective controls. Influences of exogenous melatonin varied in relation to the dose and duration of treatment and the reproductive status of the carp. However, testicular response to exogenous melatonin (at 100 µg, for 30 days) and DD in each reproductive phase was almost identical. Notably, precocious testicular maturation occurred in both DD and melatonin‐injected fish during the preparatory phase and in LL carps during the pre‐spawning phase. In contrast, testicular functions in both the melatonin‐treated and DD fish were inhibited during the pre–spawning and spawning phases, while the testes did not respond to any treatment during the post‐spawning phase. In conclusion, this study provided the first experimental evidence that melatonin plays a significant role in the regulation of annual testicular events in a sub‐tropical surface‐dwelling carp Catla catla, but the influence of this pineal hormone on the seasonal activity of testis varies in relation to the reproductive status of the concerned fish.     

Do thyroid and testis modulate the effects of pineal and melatonin on haemopoietic variables inClarias batrachus?

Involvement of pineal and its major hormone, melatonin, in the process of erythropoiesis in a freshwater catfish,Clarias batrachus has been investigated. The study was conducted during four phases, namely preparatory phase, spawning phase, postspawning phase and late postspawning phase of its annual reproductive cycle. During each phase a fish received either melatonin injections or subjected to pinealectomy. In addition, each fish in all the groups, received either iopanoic acid or cyproterone acetate or vehicle in the morning or late afternoon. Results clearly indicate that melatonin stimulates the rate of erythropoiesis inClarias batrachus. It appears that the extent of stimulation depends upon the phase of the annual reproductive cycle. However, in general, the pineal-or melatonin-induced modulation of blood variables is gonad dependent and thyroid seems to play a time of the day dependent subtle role     

Expression of melatonin synthesis genes is controlled by a circadian clock in the pike pineal organ but not in the trout

The photosensitive teleost pineal organ exhibits a daily rhythm in melatonin production. In most teleosts, including the pike, this is driven by an endogenous pineal clock. An exception is the trout, in which the pineal melatonin rhythm is a direct response to darkness. This fundamental difference in the regulation of melatonin production in two closely related species provides investigators a novel opportunity to study the molecular mechanisms of vertebrate clock function. We have studied the circadian regulation of mRNA encoding two melatonin synthesis enzymes by Northern blot analysis. These two enzymes are serotonin N-acetyltransferase (AA-NAT), the penultimate enzyme in melatonin synthesis, and tryptophan hydroxylase (TPH), the first enzyme in melatonin synthesis. A clock controls expression of both AA-NAT and TPH mRNAs in the pineal organ of pike, but not that of trout, in which the levels of these mRNAs are tonically elevated. A parsimoneous explanation of this is that a single circadian system regulates the expression of both AA-NAT and TPH genes in most teleosts, and that in trout this system has been disrupted, perhaps by a single mutation.     

Antigonadal effects of blinding in the presence of antibody against circulating melatonin

The effect of melatonin on reproductive function in the rat was studied. Reproductive organ weights and sex hormone levels were compared between sighted controls and animals which were either blinded, blinded and pinealectomized or blinded and immunized against circulating melatonin. Circulating androgens as measured by accessory sexual organ weights were significantly reduced by blinding. This effect was reversed by pinealectomy but not by immunization. Blinding also increased pineal melatonin levels but there were no differences in the plasma levels of luteinizing hormone. Circulating testosterone levels and pineal melatonin levels of immunized animals did not differ from those of blinded controls. These findings confirm reports that pineal stimulation by blinding enhances pineal melatonin content and inhibits accessory sex organ development. Circulating melatonin does not appear to be the mediator of the stimulated pineal's antigonadal effects in the rat since, in contrast to pinealectomy, neutralization of circulating melatonin failed to reverse accessory organ regression.     

Reproduction of the inshore coral trout Plectropomus maculates (Perciformes: Serranidae) from the Central Great Barrier Reef, Australia

The reproductive biology of the serranid fish Plectropomus maculatus sampled from inshore waters of the Central Great Barrier Reef was studied based on histological analyses of gonad material. This species was shown to be a monandric protogynous hermaphrodite. The process of sex change foliowed the spawning period observed during September through November. Plectropomus: maculatus showed multiple spawning during this period. Sex change followed the usual protogynous mode with degeneration of ovarian germinal tissue accompanied by proliferation of male germinal tissue in the gonad. The sex structure of the sampled population was analysed based on age and size information. The size and age of first reproduction for females was 30.0cm s. L. and 2 years of age. The size and age of sex-transition was 35.4 cm s. L. and 4.4 years of age. The sex/size and sex/age relationships indicated that sex-change can occur over a broad range of sizes and ages. The sizes and age distributions of males and females P. maculates overlapped over 38% of the length range and over 42% of the maximum age observed.     

Influence of altered photoperiods on serum melatonin and its receptors (MT1 and MT2) in the brain, retina, and ovary in carp Catla catla

Daily variation in melatonin receptor (MT1 and MT2) density in three specific tissues-brain, retina, and ovary-and its temporal relationship with serum melatonin were evaluated for the first time in a freshwater teleost, the carp Catla catla, under natural as well as altered photoperiods in different reproductive phases of the annual cycle. Cosinor analysis was used to determine rhythmic features of the serum melatonin and receptors (MT1 and MT2) in different tissues. In each photoperiodic group, irrespective of season, the daily minimum serum melatonin level was noted at midday. However, the daily peak value of melatonin varied in relation to both photo-schedules and reproductive phases. Under natural photoperiods (NPs; duration varied with seasons) and short photoperiods (SPs; light [L]:dark [D] 8:16), it occurred in the late dark phase during the preparatory phase, and at midnight in the remaining parts of the annual cycle. On the other hand, in each reproductive phase, compared to corresponding NP carp, the daily melatonin peak under long photoperiods (LPs; L:D 16:8) exhibited a phase delay of ～2-3?h (occurring during the late dark phase). The melatonin levels at each sampling point were highest during the postspawning phase and lowest during the spawning phase, irrespective of the photoperiodic history of the fish. In each tissue, Western blot analysis revealed a band at ～37?kDa and a band at ～36?kDa corresponding to the molecular weights of native MT1 and MT2 receptor proteins, respectively, with the band intensity of MT1 always being higher than that of a 36-kDa protein. The content of both melatonin receptor proteins varied significantly according to the studied tissue (being highest in the retina, intermediate in the brain, and lowest in the ovary), time in the daily cycle (peak at midnight and fall at midday), and reproductive phase in the annual cycle (highest in the spawning phase and lowest in the postspawning phase). Remarkably, no significant effects of altered photoperiod were detected on any rhythm parameters of either MT1 or MT2 in any of the studied tissues. Collectively, the results of the present study suggest a role of photoperiod in determining daily and seasonal profiles of serum melatonin, but not its receptor proteins, on the ovary or on any nongonad tissues in carp.     

Socially controlled sex-change in the half-moon grouper,Epinephelus rivulatus,at Ningaloo Reef,Western Australia

The cues controlling sex-change have been elucidated for various species of hermaphroditic fishes that inhabit coral reefs, but not for the epinepheline serranids. A male removal experiment conducted on an assemblage of the half-moon grouper, Epinephelus rivulatus, demonstrated that protogynous sex-change in this species is socially controlled, possibly by the suppressive dominance of males and a threshold sex ratio. The experiment showed that a reproductively ripe female can change sex and become a male with ripening testis within 3 weeks. However, this process can be delayed, slowed, or stopped by the presence of other males in the area.     

Inconsistent suppression of nocturnal pineal melatonin synthesis and serum melatonin levels in rats exposed to pulsed DC magnetic fields

The purpose of these experiments was to determine whether the exposure of rats at night to pulsed DC magnetic fields (MF) would influence the nocturnal production and secretion of melatonin, as indicated by pineal N-acetyltransferase (NAT) activity (the rate limiting enzyme in melatonin production) and pineal and serum melatonin levels. By using a computer-driven exposure system, 15 experiments were conducted. MF exposure onset was always during the night, with the duration of exposure varying from 15 to 120 min. A variety of field strengths, ranging from 50 to 500 μT (0.5 to 5.0 G) were used with the bulk of the studies being conducted using a 100 μT (1.0 G) field. During the interval of DC MF exposure, the field was turned on and off at 1-s intervals with a rise/fall time constant of 5 ms. Because the studies were performed during the night, all procedures were carried out under weak red light (intensity of <5 μW/cm²). At the conclusion of each study, a blood sample and the pineal gland were collected for analysis of serum melatonin titers and pineal NAT and melatonin levels. The outcome of individual studies varied. Of the 23 cases in which pineal NAT activity, pineal melatonin, and serum melatonin levels were measured, the following results were obtained; in 5 cases (21.7%) pineal NAT activity was depressed, in 2 cases (8.7%) studies pineal melatonin levels were lowered, and in 10 cases (43.5%) serum melatonin concentrations were reduced. Never was there a measured rise in any of the end points that were considered in this study. The magnitudes of the reductions were not correlated with field strength (i.e., no dose-response relationships were apparent), and likewise the reductions could not be correlated with the season of the year (experiments conducted at 12-month intervals under identical exposure conditions yielded different results). Duration of exposure also seemed not to be a factor in the degree of melatonin suppression. The inconsistency of the results does not permit the conclusion that pineal melatonin production or release are routinely influenced by pulsed DC MF exposure. In the current series of studies, a suppression of serum melatonin sometimes occurred in the absence of any apparent change in the synthesis of this indoleamine within the pineal gland (no alteration in either pineal NAT activity or pineal melatonin levels). Because melatonin is a direct free radical scavenger, the drop in serum melatonin could theoretically be explained by an increased uptake of melatonin by tissues that were experiencing augmented levels of free radicals as a consequence of MF exposure. This hypothetical possibly requires additional experimental documentation. Bioelectromagnetics 19:318–329, 1998. © 1998 Wiley-Liss, Inc.     

Diurnal and seasonal profiles of melatonin in the liver and their correlates of ovarian functions under natural photo-thermal conditions in adult carp Catla catla

Present study aimed to demonstrate daily rhythm features of hepatic melatonin concentrations in relation to ovarian functions during four reproductive phases of an annual cycle by measuring the levels of melatonin, 17-β estradiol (E2), vitellogenin (Vg) and maturation inducing hormone (MIH) in the liver and/or serum of adult carp Catla catla. Melatonin titres in liver, irrespective of reproductive phase, underwent daily variations with a peak in early dark phase and nadir at midday. However, the acrophase (Ø) of serum melatonin varied from late night in preparatory phase to midnight in the remaining parts of annual cycle. Their amplitude was highest during post-spawning phase and lowest during spawning phase. Hepatic E2 levels showed daily peak at midday and seasonal peak during pre-spawning phase. Though levels of serum Vg proteins and MIH did not exhibit daily variations, underwent seasonal changes with the highest and lowest values during spawning and post-spawning phases respectively. Hepatic melatonin titres always displayed significant negative correlation with the levels of both E2 and Vg. In essence, our study presented the first data on the daily and seasonal rhythm features of hepatic melatonin in carp and underlined their temporal relationship with the functions of ovary in any fish species.     

Melatonin rhythms in the pineal and non-pineal tissues and their physiological implications in subtropical fish

Abstract

Melatonin (N-acetyl-5-methoxy tryptamine), following discovery from the extracts of bovine pineal gland, has been detected in the pineal as well as several extra-pineal tissues/organs of different vertebrates including fish. The unique feature of melatonin in the pineal gland is its rhythmic biosynthesis and release in blood in synchronization with the environmental light-–dark cycle. Accordingly, melatonin produced in the pineal of an animal living in a changing environment is implicated to the regulation of seasonal reproduction by acting as a hormone at one or more levels of hypothalamo-hypophyseal-gonadal axis. Additionally, melatonin is known to act as a potent free-radical scavenger or antioxidant to influence maturation of oocytes. However, possible relationship between extra-pineal melatonin and seasonality of reproduction in any animal remains enigmatic. Perhaps, carp is the only known animal in which temporal patterns of melatonin levels in the serum as well as in the extracts of pineal, retina, ovary, gut, and liver have been studied in relation to the reproductive events in an annual cycle. The purpose of current review is to bring those fascinating, and arguably most important data together to underline their significance in the control of seasonal reproduction in subtropical fish in general and in carp in particular.     

Effects of Pinealectomy and Exogenous Melatonin on the Thyroid Gland Activity Varies in Relation to Reproductive Status of Male Spotted Munia (Lonchura punctulata; Aves,Passeriformes)

The purpose of this investigation was to explore whether the pineal organ and its hormone melatonin has any influence on the activity of thyroid glands, if so, how that relates to the reproductive status of a hitherto unstudied seasonally breeding wild bird. Accordingly, an identical experimental regimen was followed with adult male spotted munia (Lonchura punctulata; Passeriformes) during both its gametogenically active (August-September) and inactive (March-April) phases of the annual reproductive cycle. In either case, the levels of circulating thyroid hormones (both T₃ and T₄) and cellular characteristics of thyroid glands in groups of birds were studied following surgical removal of the pineal gland and/or daily afternoon administration of melatonin (10 μg/ 100 g body weight/ day for 30 days). The results of the same experimental schedule were found to be different depending on the sexual status of the concerned birds. During the breeding phase, pinealectomy (Px) induced significantly decreased values of T₃ and increased for T₄ along with hypertrophy of the thyroid follicular cells (TFC). The changes were reversed in melatonin treated Px birds. An increased amount of T₃ and decreased concentration of serum T₄ were also observed in melatonin injected intact birds. Conversely, the responses of TFC and of thyroid hormones in blood to either Px, or Px with melatonin, or to melatonin alone in intact munias during their inactive reproductive phase were just opposite to those noted during the breeding phase. The results of the present study suggest an influence of the pineal upon the thyroid in spotted munia. Moreover, it appears that this influence is carried out by melatonin, the action of which is reversible in relation with the gametogenic status of the concerned avian species.     

Reproductive phase depedent circadian variation in the pineal biochemical constituents of Indian palm squirrel, Funambulus pennanti

In mammals, pineal gland is intimately concerned with the co-ordination of rhythm physiology. Biochemical characteristics of pineal gland in man and other mammals may provide strong, yet sometimes elusive support for the belief in functional individuality and probable importance of this tiny gland. In seasonal breeding animals, pineal gland function is very much dependent on the reproductive status. Therefore, the aim of this experiment is to note the circadian rhythmicity of different biochemical constituents of pineal gland during active and inactive phases of reproductive cycle of a seasonally breeding rodent, F. pennanti. In the present study, pineal biochemical constituents i.e. protein and cholesterol showed higher values during daytime (1400 h). The plasma melatonin level presented two peaks during active (April; at 1800 h and 0200 h) and inactive (December; at 1400 h and 0200 h) phases of reproductive cycle. The pineal protein, cholesterol and plasma melatonin values in term of basal and peak levels were higher during the reproductive inactive/pineal active phase. Therefore, pineal--also known to have antigonadotropic properties and cholesterol which appears conjugated with pineal serotonin, presented circadian rhythmicity along with the plasma level of melatonin. This rhythmicity noted in present study was dependent on the reproductive and pineal activity status, and might be regulated by the sex steroid receptor present on the pineal gland.     

The Pineal Gland,but Not Melatonin,Is Associated with the Termination of Seasonal Testicular Activity in an Annual Reproductive Cycle in Roseringed Parakeet Psittacula krameri

The role of the pineal gland and its hormone melatonin in the regulation of annual testicular events was investigated for the first time in a psittacine bird, the roseringed parakeet (Psittacula krameri). Accordingly, the testicular responsiveness of the birds was evaluated following surgical pinealectomy with or without the exogenous administration of melatonin and the experimental manipulations of the endogenous levels of melatonin through exposing the birds to continuous illumination. An identical schedule was followed during the four reproductive phases, each characterizing a distinct testicular status in the annual cycle, namely, the phases of gametogenic quiescence (preparatory phase), seasonal recovery of gametogenesis (progressive phase), seasonal initiation of sperm formation (pre‐breeding phase), and peak gametogenic activity (breeding phase). In each reproductive phase, the birds were subjected to various experimental conditions, and the effects were studied comparing the testicular conditions in the respective control birds. The study included germ cell profiles of the seminiferous tubules, the activities of steroidogenic enzymes 17β‐hydroxysteroid dehydrogenase (17β‐HSD), and Δ⁵3β‐hydroxysteroid dehydrogenase (Δ⁵3β‐ HSD) in the testis, and the serum levels of testosterone and melatonin. An analysis of the data reveals that the pineal gland and its hormone melatonin may play an inhibitory role in the development of the testis until the attainment of the seasonal peak in the annual reproductive cycle. However, in all probability, the termination of the seasonal activity of the testis or the initiation of testicular regression in the annual reproductive cycle appears to be the function of the pineal gland, but not of melatonin.     

Pineal and serum melatonin at midday and midnight following pinealectomy or castration in male rats.

Melatonin has been extractkd by chloroform from rat serum and quantified by the Rana pipiens tadpole bioassay. Like pineal melatonin, serum melatonin was high at mid-dark and low at mid-light. This finding suggests a diurnal rhythm of serum melatonin in the rat. Serum samples obtained at mid-dark had melatonin levels ranging from 0.02 to 0.05 ng per ml of serum. When rats were pinealectomized, melatonin activity in the serum was abolished. Thus, rat serum melatonin may originate primarily from the pineal. There was no significant change in pineal melatonin content nor in serum melatonin concentrations 7 or 12 days following orchidectomy.