Seasonality of pineal gland activity and immune functions in chickens

The immunomodulatory action of melatonin in different animal species is already well known, although the mechanism(s) by which the indoleamine influences the immune system have yet to be fully elucidated. Previously, we have shown both anti-inflammatory and opioid-mediated influence of exogenous melatonin on thioglycollate-induced peritonitis in young chickens. In the present study, the kinetics of peritonitis and splenocyte proliferation were compared in chickens reared in both seasons under the same L:D 12:12 conditions. These two aspects of the immune response were correlated with the diurnal rhythm of pineal gland function, measured by the activity of N-acetyltransferase (NAT), a key enzyme in melatonin biosynthesis. The results revealed seasonal changes in the circadian rhythm of pineal NAT activity occurring in parallel to the natural local geophysical seasons. These changes appeared to influence the development of peritonitis and splenocyte responsiveness to mitogenic stimulation in vitro. Moreover, the existence of bidirectional communication between the pineal gland and the activated immune system was supported by the decreased activity of pineal NAT in chickens with peritonitis compared with control birds.     

Influence of prolactin on pineal gland activity in chicks

The aim of the present investigation is to ascertain the effects of prolactin on the pineal gland activity in chicks. Ovine prolactin treatment at all the dosages (total dosage: 15, 50, 100 IU per bird in 10 days) increased the mitotic frequency and nuclear size in the pineal gland of chicks. The rate of increase of these manifestations was seen in proportion to the amount of hormone administered. It is suggested that prolactin can perhaps stimulate the pineal gland function in chick and that it has a dose-dependent action in this gland.     

Thymidine kinase activity of the chick pineal gland

Pineal thymidine kinase activity of 1-week-old chicks in situ varied significantly throughout the day. However, the circadian rhythm of thymidine incorporation seen with cultured chick pineal glands was not accompanied by variations in level of thymidine kinase activity in vitro. Thus the circadian rhythm in rate of cumulative incorporation of thymidine by cultured chick pineal glands is not determined by a rhythm in rate of the first reaction of the complex series of reactions by which thymidine is incorporated into DNA.     

Light-induced changes in pineal gland N-acetyltransferase activity: developmental aspects

In adult rats, light acting via a retino-pineal gland neural pathway influences pineal gland biochemistry in two ways: (1) it entrains endogenous circadian rhythms in melatonin biosynthesis to the environmental photoperiod and (2) exposure to even very brief periods of light during the nighttime rapidly suppresses the high levels of nocturnal melatonin production. The present studies were undertaken to determine precisely when photic stimulation first influences the enzymic activity of N-acetyltransferase (NAT), the pineal gland enzyme which rate-limits the overall biosynthesis of the hormone melatonin, and to examine some of the cellular mechanisms which might mediate light-induced effects in neonatal animals. Rats of different ages were either killed during the light phase or were exposed to darkness or light for 1 min during the dark phase of the lighting cycle, returned to their litters in darkness for 30 min and then killed. Pineal gland NAT activity in animals nocturnally exposed to 1 min of light was suppressed in animals 6 days of age or older. Nocturnal light exposure did not suppress enzyme activity in 3- to 5-day-old rats, even though these animals displayed clear light:dark differences in pineal gland NAT activity. Nocturnal light exposure also did not suppress nighttime levels of NAT activity in 7-day-old animals who had been bilaterally enucleated, suggesting that this effect is retinally mediated. Pretreatment of 7-day-old animals with the beta-noradrenoceptor agonist drug, isoproterenol, prevented the nocturnal light-induced suppression of NAT activity.(ABSTRACT TRUNCATED AT 250 WORDS)     

Light-induced suppression of nocturnal serotonin N-acetyltransferase activity in chick pineal gland and retina: A wavelength comparison

Abstract: Effects of white and monochromatic (blue—434 nm, green—548 nm, and red—614 nm) lights on the nighttime retinal and pineal NAT activity were examined in chicks. The potency of the tested lights to suppress NAT activity was similar for the retina and pineal gland, with a following rank order: white > green > blue > red. The studied tissues of chick were far less sensitive to pulses of monochromatic light than the rat pineal gland. The potency of light to decrease pineal NAT activity of rat was: white > green >> blue > red. In chicks, the suppression of the nocturnal NAT activity produced by a short 5-min pulse of monochromatic light was completely reversible in the pineal gland, and partially reversible in the retina. Our data suggest the existence of some differences between birds and mammals in terms of sensitivity and mechanisms involved in the light-induced suppression of melatonin biosynthesis.     

A renin-like enzyme in luteal tissue

The aim of this study was to identify immunologically and biologically a renin-like enzyme (RLE) in rat corpora lutea (CL). The biological activity of partially purified extracts of CL was tested in vivo by injection into anesthetized pentolinium-treated rats, obtaining a pressor response similar to renal renin. The enzyme activity in vitro was inhibited to about 50% by pretreatment with a specific antibody against renal renin. When the extracts were incubated with angiotensinogen, the product was inhibited mainly by angiotensin I antibody. The fact that there was no change in RLE content in 24 or 48 h nephrectomized rats, suggested the idea of a local production rather than an active blood renin sequestration.     

The pineal gland and mammalian photoperiodism

The mammalian pineal gland appears to be a major endocrine component in the regulation of photoperiodic responses. The circadian pattern of secretion of the pineal hormone, melatonin, is regulated by the nervous system. Changes in photoperiod, acting via the nervous system, alter the temporal pattern of melatonin secretion. The changes in secretion pattern convey information about daylength from neural components of the circadian system to the reproductive system, and probably to other physiological systems.     

Methodologic features of determining renin-like activity in human arteries]

Human arterial tissue was shown to have renin-like activity. For its determination the authors propose methodological approaches. The optimum pH of renin activity in the vascular wall was found to be 5.8-6.0. The renin-like enzyme was ascertained to be present in the vascular wall as inactive and to be activated by trypsin. The renin activity was compared in the human vascular wall and human plasma. Their optimum pHs were nearly identical. The methodological features determined in this paper allowed one to differentiate the true renin-like activity from the activity of acid proteases.     

Alpha-methyltryptophan metabolism in rat pineal gland and brain

Alpha-methyltryptophan (AMTP), a synthetic amino acid, is metabolized by the rat in vivo to alpha-methylserotonin (AM5HT), which appears in the pineal gland just as it does in the brain. Pineal AM5HT assumes the same diurnal rhythm as serotonin does in control animals. Administration of AMPT results in a decrease of the serotonin content of the pineal gland, but not of its melatonin content. Pharmacological evidence indicates that the uptake of AMTP into the gland is influenced by noradrenergic innervation. No evidence was obtained for formation in vivo of alpha-methyl-N-acetylserotonin or the alpha-methyl analogue of melatonin.     

Somatostatin and prosomatostatin immunoreactive nerve fibers in the bovine pineal gland.

An immunohistochemical study of the bovine pineal gland was performed by using polyclonal rabbit antisera raised against synthetic peptide fragments corresponding to the amino acid sequences of somatostatin-14, somatostatin-28, somatostatin-28 (1-12), and prosomatostatin (20-36). Immunoreactive nerve fibers were demonstrated in the pineal gland with all four antisera. The nerve fibers were located throughout the gland, both perivascularly and intraparenchymally, with the highest density of fibers in the proximal part, close to the pineal stalk. A number of immunoreactive nerve fibers were also found in the habenular area and stria medullaris projections. Some of the fibers in the stria medullaris projections could be followed into the pineal gland via the rostral part of the pineal stalk. A few immunoreactive nerve fibers were present in posterior commissure, in the subcommissural organ and pretectal area. Thus, the present study shows that the bovine pineal gland is innervated by nerve fibers containing all four sequences of prosomatostatin. The anatomical location of the somatostatin-immunoreactive nerve fibers in the bovine pineal gland indicates that the perikarya of some of these nerve fibers are located in the brain.     

Daily rhythm of aspartate aminopeptidase activity in the retina, pineal gland and occipital cortex of the rat.

The levels of specific soluble aspartyl aminopeptidase activity were assayed in retina, occipital cortex, anterior pituitary, posterior pituitary, pineal gland and serum of adult male rats, using Asp-2-naphthylamide as substrate, in a 12:12 h light:dark cycle (7-19 h light). Significant diurnal variations appeared in retina, pineal gland, occipital cortex and serum. In addition, different patterns of diurnal variation of the enzymatic activity were observed in the tissues analyzed. A regular increase of the activity was noticed at the end of the dark period in all the tissues as a common feature, except in serum, in which the enzymatic activity reached a peak in the middle of the light period, decreasing progressively during the dark hours. After the last hours of darkness, the pattern of variation in the activity differed in each tissue. These diurnal variations in aspartyl aminopeptidase activity could reflect the functional status of its putative endogenous substrates, such as angiotensin II, and it may also suggest the existence of differential regulatory mechanisms associated with each location.     

GABAergic signaling in the rat pineal gland

Pinealocytes secrete melatonin at night in response to norepinephrine released from sympathetic nerve terminals in the pineal gland. The gland also contains many other neurotransmitters whose cellular disposition, activity, and relevance to pineal function are not understood. Here, we clarify sources and demonstrate cellular actions of the neurotransmitter γ‐aminobutyric acid (GABA) using Western blotting and immunohistochemistry of the gland and electrical recording from pinealocytes. GABAergic cells and nerve fibers, defined as containing GABA and the synthetic GAD67, were identified. The cells represent a subset of interstitial cells while the nerve fibers were distinct from the sympathetic innervation. The GABA_A receptor subunit α1 was visualized in close proximity of both GABAergic and sympathetic nerve fibers as well as fine extensions among pinealocytes and blood vessels. The GABA_B1 receptor subunit was localized in the interstitial compartment but not in pinealocytes. Electrophysiology of isolated pinealocytes revealed that GABA and muscimol elicit strong inward chloride currents sensitive to bicuculline and picrotoxin, clear evidence for functional GABA_A receptors on the surface membrane. Applications of elevated potassium solution or the neurotransmitter acetylcholine depolarized the pinealocyte membrane potential enough to open voltage‐gated Ca²⁺ channels leading to intracellular calcium elevations. GABA repolarized the membrane and shut off such calcium rises. In 48–72‐h cultured intact glands, GABA application neither triggered melatonin secretion by itself nor affected norepinephrine‐induced secretion. Thus, strong elements of GABA signaling are present in pineal glands that make large electrical responses in pinealocytes, but physiological roles need to be found.     

Swimming depresses nighttime melatonin content without changing N-acetyltransferase activity in the rat pineal gland

Recently, it was shown that a 1.5-ml subcutaneous saline injection depressed N-acetyltransferase (NAT) activity and melatonin content in the rat pineal gland at night. The present studies were undertaken to determine if another perturbation, swimming, could duplicate this response. Rats swam at 23.10 h (lights out at 20.00 h) for 10 min and were killed 15 and 30 min after the unset of swimming. Pineal NAT activity was found to be unaffected while melatonin content was depressed dramatically. Hydroxyindole-O-methyltransferase (HIOMT) activity as well as the content of serotonin (5HT), 5-hydroxytryptophan (5HTP) and 5-hydroxyindoleacetic acid (5HIAA) were not changed by this treatment. In a second study, pineal melatonin again was depressed without a concomitant drop in NAT activity. Mean serum melatonin at 15 min after onset of swimming was increased although the rise was not statistically significant. In the final study, it was found that NAT activity was slightly increased in intact rats and unchanged in adrenalectomized rats at 7 min after swimming onset. At 15 min both intact and adrenalectomized animals had NAT activity values similar to those of controls. Pineal melatonin content in intact and adrenalectomized rats plummeted to 50% of control values at 7 min and fell further to 25% at 15 min. While the rate of melatonin synthesis was not directly measured, lack of change in the activities of the enzymes involved in melatonin synthesis and the contents of two melatonin precursors suggests that swimming depresses pineal melatonin content by enhancing melatonin efflux from the gland.