Circadian rhythms of indoleamines and serotonin N-acetyltransferase activity in the pineal gland

Conclusion The circadian rhythm of melatonin synthesis in the pineal glands of various species has been summarized. The night-time elevation of melatonin content is in most if not all cases regulated by the change of N-acetyltransferase activity. In mammals, the N-acetyltransferase rhythm is controlled by the central nervous system, presumably by suprachiasmatic nuclei in hypothalamus through the superior cervical ganglion. In birds, the circadian oscillator that regulates the N-acetyltransferase rhythm is located in the pineal glands. The avian pineal gland may play a biological clock function to control the circadian rhythms in physiological, endocrinological and biochemical processes via pineal hormone melatonin.     

In vitro effects of steroid hormones on arylalkylamine N-acetyltransferase (AA-NAT) activity in the pineal of fish, Clarias gariepinus (Burchell, 1822) during different phases of breeding cycle

In vitro effects of gonadal hormones (testosterone, 17beta-estradiol estriol and estrone) and corticosteroid hormones (corticosterone and cortisol) were studied on arylalklyamine N-acetyltransferase (AA-NAT) activity in the pineal organ of the fish, C. gariepinus during quiescent, progressive, breeding and regressive phases of its annual breeding cycle. The pineals were collected under dim red light, maintained in organ culture for 7 hr and incubated with three concentrations (10(-6), 10(-5) and 10(-4) M) of hormones for 6 hr. The treatments with gonadal hormones and corticosteroid hormones inhibited pineal AA-NAT activity in a dose-dependent manner during all the phases of the breeding cycle. AA-NAT activity was comparatively more sensitive to the inhibitory effects of the gonadal hormones during the regressive phase and less sensitive during the quiescent phase. Further, the enzyme activity was more sensitive to the inhibitory effects of corticosteroid hormones (corticosterone and cortisol) during the breeding phase and less sensitive during the quiescent phase. These findings seem to suggest that gonadal hormones and corticosteroid hormones have direct inhibitory influence on AA-NAT activity and, hence melatonin synthesis in the photoreceptive pineal organ of C. gariepinus.     

Effects of glucocorticoids on plasma levels of thyroid hormones (T4 and T3) and testicular activity in catfish, Clarias gariepinus during different phases of annual breeding cycle

Effects of short-term administration of corticosterone and cortisol on plasma levels of thyroid hormones, gonado-somatic index and testicular histology have been reported in catfish, Clarias gariepinus during different phases of its breeding cycle. Corticosterone administration had no significant effect on plasma levels of T4, T3 and T3/T4 ratio, irrespective of doses and phases of breeding cycle. However, 5 microg dose of cortisol significantly increased plasma levels of T3 and the T3/T4 ratio during quiescent and regressive phases, while it significantly decreased plasma levels of T4 during progressive phase. During breeding phase, 2 microg and 5 microg doses of cortisol significantly decreased plasma levels of T4 and T3, respectively, while 5 microg dose of cortisol alone reduced T3/T4 ratio. Irrespective of phases of annual breeding cycle and doses, short-term administration of corticosterone and cortisol had no significant effect either on GSI or testicular histology. These findings suggest that corticosterone is ineffective in stimulating plasma levels of thyroid hormones, while cortisol, depending on dose and phase/season, may differentially increase, decrease or have no effect on plasma levels of thyroid hormones in C. gariepinus.     

Effect of ammonia-stress on the total autolytic levels of proteins in tissues of an air-breathing fish,Channa punctatus (Bloch)

G season-specific effect of a sub-lethal ambient ammonium chloride concentration on the total autolysis of protein in different tissues of the Indian air-breathing murrel,Chauna punctatos (Bloch), has been demonstrated. While its effect on the autotytie levels of protein of different tissues (except white muscle) was marginal in the winter-adapted fish, the same in the summer-adapted fish was more pronounced. In general a reduction in the autolysic levels of the tissue protein of the ammonium chloride-stressed fish was observed. In certain tissues like white muscle of an ammonium chloride-stressed fish and gill of the ammonium hydroxide-stressed fish, augmented levels of autolysis were noticed. The present findings suggest that the accumulation of amino acids in different tissues of the ammonia-stressed fish, as observed earlier, can be explained best by mechanisms other than ammonia induced increased breakdown of proteins.     

Role of thyroidal and testicular hormones in regulation of tissue respiration in male air-breathing fish, Clarias batrachus (Linn.)

In vivo and in vitro effects of thyroidal hormones (MIT, DIT, T3, T4), propyl thiouracil (PTU), testosterone and cyproterone acetate were studied on the rate of tissue (liver, muscle, kidney and brain) respiration of adult male C. batrachus during winter and summer/rainy seasons. Monoiodotyrosine (MIT) and diiodothyrosine (DIT) increased the respiratory rate in a dose-dependent and temperature-independent manner. Triiodothyronine (T3) and thyroxine (T4) stimulated tissue respiration during summer/rainy months but not during winter. PTU decreased tissue respiration during summer/rainy season and also at simulated low temperature. Testosterone invariably stimulated the rate of respiration of the tissues, while in vivo treatment with cyproterone acetate significantly decreased the metabolic rate of all the tissues. The findings suggest that in C. batrachus MIT and DIT may be more important than T3 and T4 at low temperature, endogenous thyroid hormones are involved indirectly in energy metabolism even during winter/at low temperature and testicular hormones are actively involved in the respiration.     

Effect of GnRHa, pimozide and Ovaprim on ovulation and plasma sex steroid hormones in African catfish Clarias gariepinus

Nine groups each of four fish were injected with a single intramuscular dose of the following preparations: Physiological saline (0.9% NaCl) as a control group, 0.5 ml kg⁻¹ Ovaprim, 20 and 40 μg kg⁻¹ BW of GnRHa, 8 and 16 mL kg⁻¹ pimozide tablets and the following combination of GnRHa with pimozide (GP): 20 μg + 4 mg, 30 μg + 8 mg and 40 μg + 16 mg kg⁻¹ BW. The primary oocyte diameter (POD) before hormone administration ranged from 943.3 to 1071.0 μm. The latency periods (LP) were in the range of 9.0 to 12.0 h after injection. The highest ovulation ratio (OR) was observed in groups Ovaprim, GP(30 + 8) and GP(40 + 16). Other treatments were effective for ovulation, the ovulation ratio in Groups G(40) and GP(20 + 4) were significantly higher than G(20) treatment. The ovulation index (OI) was in the range 62 to 77% and showed significant differences among groups. There was no significant difference in fertilization ratio (FR) among Ovaprim, GP(30 + 8) and GP(40 + 16) groups, while there were significant difference between the previous group and G(20) and G(40) groups. Control, P8, P16 showed negative results in all the parameters LP, OED, OR, OI and FR. Levels of sex steroids were analyzed on 6 and 12 h after initiation of treatments. A significant increase in plasma E₂ with GP(30 + 8) injection was observed 6 and 12 h after injection, while there were no significant increase between all the other groups 6 h after injection. Treatments with GP(20 + 4) resulted in a significant increase in plasma T concentration in females compared with control after 6 h. In contrast, plasma T and E₂ concentrations were lower during the combined GP(20 + 4), GP(30 + 8) and GP(40 + 16) after 12 h than after 16 h of injection. The combined treatments (GnRHa + PIM) are better compared with Ovaprim which gave the same results, they have some advantages, such as reliable response and low cost. Ovaprim is more than 3 to 5-fold of the cost of (GnRH + PIM). Therefore, this method could be useful tool for commercial catfish breeders to ensure spawning success.     

Cytologic variation in the pineal organ of roseringed parakeets (Psittacula krameri) with respect to artificial photoperiods and annual cyclicity in testicular functions

Abstract

Adult male roseringed parakeets were transferred to 16L: 8D (LP), or 8L: 16D (SP) for 45 or 90 days on four particular dates corresponding to the different phases of an annual testicular cycle and the cytological responses of the pineal were studied comparing them with the features in respective natural photoperiodic (NP) birds. Different cytological characteristics including the values of nuclear diameter in pinealocytes indicated that LP for 45 days during the pre‐breeding phase and for 90 days during each phase resulted in an increased pineal activity, while SP for 45 and 90 days induced inhibitory responses of the pineal during each, but not the progressive phase of the annual testicular cycle. During the latter phase, none of the artificial photoperiodic schedules, other than LP for 90 days, influenced the cytological features of the pineal. The results suggest that the photosensitivity of the pineal in these parakeets varies in relation to the testicular functions in an annual testicular cycle, but the seasonal pattern of photoperiodic response of the pineal and that of the previously studied testes in the same birds does not seem to be identical.     

In vitro effects of 5-hydroxytryptophan, indoleamines and leptin on arylalkylamine N-acetyltransferase (AA-NAT) activity in pineal organ of the fish, Clarias gariepinus (Burchell, 1822) during different phases of the breeding cycle

Arylalkylamine N-acetyltransferase (AA-NAT) is the rate-limiting enzyme of melatonin biosynthetic pathway. In vitro effects of 5-hydroxytryptophan (5-HTP) and indoleamines (serotonin, N-acetylserotonin and melatonin) were studied on AA-NAT activity in the pineal organ of the fish, C. gariepinus during different phases of its annual breeding cycle. Further, in vitro effects of leptin on AA-NAT activity in the pineal organ were studied in fed and fasted fishes during summer and winter seasons. Treatments with 5-HTP and indoleamines invariably stimulated pineal AA-NAT activity in a dose-dependent manner during all the phases. However, leptin increased AA-NAT activity in a dose-dependent manner only in the pineal organ of the fed fishes, but not of the fasted fishes irrespective of the seasons.     

The relationship between basal metabolic rate (BMR) and concentrations of plasma thyroid hormones in fasting cockerels

A correlation between the Basal Metabolic Rate (BMR) and the level of rT3, and occasionally between BMR and T3 or T4 was found in 12 month fasting cockerels. The birds were fasted for 48 hrs and BMR was measured eight times (before fasting, at 6, 12, 24, 30, 36, and 48 hrs of fasting, and 4 hours after fasting). Blood samples for plasma collection were taken immediately after measuring the BMR. During starvation a decrease in BMR was observed. After refeeding BMR returned to the starting level. The decrease in BMR was accompanied by an increase in rT3 and T4 plasma levels. Between BMR and levels of T4 and rT3 negative coefficients of correlation were observed (r = -0.20 and r = -0.42, respectively). Contrary to this, the T3 level declined and was correlated with BMR (r = 0.62). After refeeding, the T3 level rapidly increased against the control value. Moreover, a high coefficient of correlation (r = -0.39) was found between the level of T3 and rT3. The data show that the reduction in plasma T3 level and increase in the rT3 one during starvation may be due to inhibition of deiodination of T4 to T3, since rT3 is a competitive inhibitor of this reaction. The presented results support the suggestion that in birds T3 is the metabolically active thyroid hormone, and rT3 antagonizes this effect.     

The effects of sex hormones,prolactin, and chorionic gonadotropin on pineal electrical activity in guinea pigs

Microelectrophoretic application of sex hormones onto pineal cells in guinea pigs has shown different responses in pregnant females as compared to males. In pregnant females estrone caused excitation in 74% of the cells tested, while progesterone and testosterone, prolactin, and HCG were inhibitory in a majority of the cells. In contrast, in males estrone caused excitation of only 19% but inhibition of 37%. A smaller percentage of cells was inhibited by progesterone, while the predominant response to testosterone was excitation. These results suggest that the pineal gland may be under a feedback control.This work was supported by the Volkswagenwerk-Stiftung, Grant I/35472.Visiting scientist supported by grant from the Deutsche Forschungsgemeinschaft and the Royal Society.     

Diurnal rhythms of plasma growth hormone,somatostatin, thyroid hormones,cortisol and glucose concentrations in rainbow trout,Oncorhynchus mykiss,during progressive food deprivation

Abstract

The diurnal patterns of changes in plasma cortisol, growth hormone (GH), somatostatin‐14 (SRIF), thyroid hormones (L‐thyroxine, T₄ and triiodo‐L‐thyronine, T₃) and glucose were investigated in rainbow trout, Oncorhynchus mykiss, both during single meal‐feeding and during a progressive fast of 13 weeks. All measured variables exhibited a diurnal pattern in fed rainbow trout, most of which appeared to be correlated with the time of feeding (30–60 min after the onset of light), while additional changes, associated with the scotophase were also found for cortisol. Although fasting had no affect on mean daily plasma cortisol or SRIF concentrations, there was a progressive increase in mean daily plasma GH concentrations and a progressive decrease in mean daily plasma thyroid hormone and glucose concentrations associated with fasting. However, for GH, significant changes were not evident until week 10 of the fast. In addition, fasting appeared to phase‐shift the diurnal patterns of plasma GH, cortisol and glucose concentrations; the consequence of such a shift is that conclusions as to the effects of fasting, if based on a single time of sampling, may be flawed.     

Zinc supplementation on serum levels and hepatic conversion of thyroid hormones in obese (ob/ob) mice

The supplemental effects of zinc on thyroid status in obese (ob/ob) mice were studied. Four-week-old obese mice and their lean controls were fed either a basal diet or a zinc-supplemented diet (200 mg/kg diet) for 8 wk. Following the 8-wk basal diet, obese mice had lower serum T4 values, as well as hepatic T4 and T3 values, than lean mice (p < 0.05). A significant decrease in hepatic 5′-deiodinase activity was also observed in obese mice. Dietary zinc supplementation significantly reduced serum T4 levels in both the obese and lean mice. However, the zinc-supplemented effects on diminishing hepatic T4 and T3 values, as well as on 5′-deiodinase activities, were found only in obese mice (p < 0.05). Furthermore, the 5′-deiodinase activities in hepatic microsomal pellets after incubation with various zinc concentrations (0.5, 1.0, and 2.5 mM) were also examined. The 5′-deiodinase activities, in hepatic samples from all mice, were significantly attenuated by zinc treatments. However, this effect was more predominant in obese mice following the addition of 0.5 mM zinc. This study suggests that a lower hepatic 5′-deiodinase activity, resulting from a higher zinc level, might be related to abnormal energy metabolism in theob/ob mice.     

Diel rhythms of adipokinetic hormone, fat body response, and haemolymph lipid and sugar levels in the house cricket

Abstract. The adipokinetic hormone (Grb-AKH) content in the corpora car-diaca of the house cricket, Acheta domesticus , varied during a day with two peaks in the scotophase and one peak in the photophase. There were two distinct peaks of total lipid concentration in the blood, one early in the photophase and the other early in the scotophase. Fat body sensitivity to adipokinetic hormone also varied in close synchrony with the lipid rhythm. It was not possible to attribute the rhythm of blood lipid titre unequivocally to either the rhythm of adipokinetic hormone content in the corpora cardiaca or to the rhythm of sensitivity of the fat body to the hormone. In adult crickets the blood carbohydrate titre had two peaks in adult females, one towards the end of scotophase and another in the late photophase (c. 12h apart), but a single peak at the end of scotophase was apparent in last instar larvae. The blood carbohydrate rhythm persists in DD and is therefore endogenous. Carbohydrates were not mobilized by the Grb-AKH, therefore could not be involved in the blood carbohydrate rhythm. Exposure to various day lengths caused shifts in the patterns of the carbohydrate rhythms, but imposition of a cyclic temperature regime had no effect on the rhythm.     

Changes in plasma thyroid hormone levels in pink salmon, Oncorhynchus gorbuscha, during their spawning migration in the Fraser River (Canada)

Plasma thyroid hormone levels were measured in pink salmon, Oncorhynchus gorbuscha , during their spawning migration in the Fraser River, British Columbia. The plasma levels of both l-thyroxine (T₄) and triiodo-l-thyronine (T₃) were significantly higher in males than in females. In both sexes the hormone levels were maintained, or increased somewhat, during the early stages of migration, but fell thereafter. In females the plasma T₄ and T₃ levels of salmon collected on the spawning grounds were at or below detectable levels of the assays. The changes in thyroid hormone levels are correlated with changes in plasma insulin, gonadotropin, gonadal steroid hormones, cortisol and vitellogenin levels measured in the same specimens.     

Nutritive value of dried lactic acid fermented fish silage and soybean meal in dry diets for juvenile catfish, Clarias gariepinus (Burchell, 1822)

Dry diets containing either fish meal (C-FM) or dried fermented fish silage and soybean meal blend (1:1, ww⁻¹) (C-FS) as the sole protein source, were fed to triplicate groups of juvenile Clarias gariepinus (10.8 ± 0.3 g) at 5% body weight per day for 70 days. Catfish fed the C-FS diet showed reduced (P < 0.05) growth rate, feed conversion, protein efficiency and digestibility. Lower amounts of available amino acids in the C-FS diet resulted in inferior nutritive value for catfish growth than in the C-FM diet. Postprandial changes in plasma amino acids showed similar patterns in both diet treatments, but the maximal mean levels attained for the C-FS diet were correspondingly lower and occurred earlier than with the C-FM diet. There were no effects of feeding C-FS diet on the hepatosomatic index but carcass analysis showed that body protein deposition was lower (P < 0.05). Differences in haematocrit, haemoglobin content and liver histology were demonstrated but were not pathological. Lower digestible energy of C-FS diet also contributed to the poor performance of catfish in this treatment. Results of this study indicate that C. gariepinus cannot metabolize protein from co-dried fish silage as efficiently as fish meal protein when used as the sole dietary protein.     

Fine structure of the pineal organ in the troglobytic fish,Typhlichthyes subterraneous (Pisces: Amblyopsidae)

Summary The pineal organ of the blind, cave-dwelling fish, Typhlichthyes subterraneous, was examined with both light and electron microscopes. Like the eyes, the pineal in this troglobytic species was found to be regressed. Two cell types, photoreceptor and supportive cells, were described in the pineal epithelium. Although ganglion cells were not identified, small, unmyelinated nerve fibers were present. The photoreceptor cells had degenerated outer segments. Accordingly, it was suggested that the pineal in this species is not likely to function in photoreception. However, the presence of well developed Golgi bodies, clear and dense-cored vesicles, variable amounts of rough endoplasmic reticulum and glycogen particles indicated that both cell types are metabolically active and may play a role in secretion.     

A karyological analysis of the artificial hybridization between Clarias gariepinus (Burchell, 1822) and Heterobranchus longifilis Valenciennes, 1840 (Pisces; Clariidae)

A karyological analysis of an artificial hybridization (reciprocal crosses) between two African clariid catfish, Clarias gariepinus (Burchell, 1822) and Heterobranchus longifilis Valenciennes, 1840, was performed. C. gariepinus has a standard karyotype of 2 n = 56, while H. longifilis has 2 n = 52. The hybrids revealed an intermediate karyotype (2 n = 54), and it appears as if they have totalized the haploid chromosome number of both parental species, excluding gynogenesis or androgenesis. The hybrid karyotype is considered as aneuploid, although the hybrids proved to be fertile. No variation was found in the hybrids karyotypes.     

Age-related changes in diurnal rhythms and levels of gonadotropins, testosterone, and inhibin B in male rhesus monkeys (Macaca mulatta)

Testosterone shows circadian rhythms in monkeys with low serum levels in the morning hours. The decline relies on a diminished frequency of LH pulses. Inhibin B shows no diurnal patterns. In elderly men, the diurnal rhythm of testosterone is blunted and inhibin levels fall. Here we explore whether aging exerts similar effects in the rhesus monkey. We collected blood samples from groups of young (6-9 yr) and old (12-16 yr) male rhesus monkeys at 20-min intervals for a period of 24 h under remote sampling via a venous catheter. We determined moment-to-moment changes in plasma levels of testosterone, FSH, and LH by RIA, and of inhibin B by ELISA. We found significant diurnal patterns of testosterone in both groups. The circadian rhythm in testosterone was enhanced in older monkeys. Testosterone levels and pulse frequencies dropped significantly below those of young monkeys during midday hours. Diminished pulse frequency of LH appeared to be responsible for the midday testosterone decrease in old monkeys, while LH and testosterone pulse frequency did not change in young monkeys at corresponding time points. Old monkeys showed extended periods of LH-pulse quiescence in the morning and midday hours. Inhibin B and FSH levels were generally lower in old monkeys compared with the young group, but neither inhibin B nor FSH showed circadian rhythms. We conclude from these data that old rhesus monkeys have a more prominent circadian rhythm of LH and testosterone resulting from an extended midday period of quiescence in the hypothalamus-pituitary-gonadal axis.     

Effects of time of l-ornithine administration on the diurnal rhythms of plasma growth hormone,melatonin, and corticosterone levels in mice

The synthesis and secretion of many hormones such as growth hormone (GH), melatonin, and corticosterone, exhibit temporal variations over each day and night. Oral administration of several nutritional factors, including l-ornithine, modulates these hormonal secretions and induces an acute increase in plasma GH levels. However, the impact of l-ornithine on the diurnal rhythms of hormone secretion remains unclear. In this study, we evaluated whether the diurnal rhythms of plasma GH, melatonin, and corticosterone secretion were altered by the daily administration of l-ornithine as well as the timing of the administration, in CBA/N mice. Our results showed that the plasma GH levels that peaked at light phase were amplified by l-ornithine (500?mg/kg) administered at Zeitgeber time (ZT) 22, but not at ZT10. Additionally, l-ornithine (1000?mg/kg) administered at ZT22 advanced the onset of the nocturnal rise of melatonin, which resulted in the elongation of the melatonin peak. On the other hand, l-ornithine (500 and 1000?mg/kg) administered at ZT10, but not at ZT22, suppressed the diurnal rhythm peaks of plasma corticosterone. The effects of l-ornithine on plasma GH rhythms lasted for at least 2 days after cessation of the daily administration. Running wheel activity during the active phase was slightly elevated by l-ornithine administration at ZT22, but the overall patterns were only slightly affected. l-Ornithine levels in the plasma and hypophysis after a single administration of l-ornithine at ZT22 were lower than those after administration at ZT10, suggesting that the metabolic rate of l-ornithine differs between day and night. In conclusion, our data suggest that a daily administration of l-ornithine regulates the diurnal rhythms of GH, melatonin, and corticosterone in a manner dependent on administration time, which might be related to the diurnal rhythms of l-ornithine metabolism.