Effects of photoperiod and melatonin on the development of growth hormone cells and the pituitary-adrenal axis in the Djungarian hamster, Phodopus sungorus.

The development of GH cells and the pituitary-adrenal axis was morphologically examined in male Djungarian hamsters (Phodopus sungorus) exposed to short days and those kept under long days and receiving daily afternoon injections of melatonin, from the time of weaning (20 days) until 100 days of age. The postnatal increase in area of ACTH cells under long days was inhibited in short-day-exposed or melatonin-treated animals. It was suggested that a short photoperiod may suppress, via melatonin, the development of ACTH cells. GH cells were not affected by age, photoperiod or exogenous melatonin. Under long days, the zona fasciculata decreased in volume with age, while the zona reticularis increased. Such changes in the volumes of these adrenocortical zones were depressed under short days. In addition, the volumes of the zona fasciculata and zona reticularis in long-day-housed animals became respectively larger and smaller subsequent to orchidectomy and melatonin administration. These results suggest that fasciculata cells in deeper levels become progressively differentiated into reticularis cells, that short photoperiod inhibits development of both zonae, and that such an inhibition is caused mainly by the decreased secretion of androgens.

Central GABAergic innervation of the mammalian pineal gland: a light and electron microscopic immunocytochemical investigation in rodent and nonrodent species.

Light and electron microscopic immunocytochemical observations were made to demonstrate central pinealopetal fibers immunoreactive for gamma-aminobutyric acid (GABA) and synapses between their terminals and pinealocytes in the pineal gland of four rodent (Wistar-King rat; mouse; Syrian hamster, Mesocricetus auratus; Hartley strain guinea pig) and one nonrodent (tree shrew, Tupaia glis) species. GABA-immunoreactive myelinated and unmyelinated fibers and endings were found in the parenchyma of the pineal gland of all the animals examined. In the rodent species, GABAergic fibers were mainly found in the intermediate and proximal portions of the pineal gland and were nearly or entirely absent in the distal portion of the gland. Abundant GABAergic fibers were evenly distributed throughout the gland of the tree shrew. In all the animals, the habenular and posterior commissures contained abundant GABA-positive fibers, and some of them were followed to the pineal gland. GABA-positive endings made synaptic contact with pinealocytes, occasionally in mice and guinea pigs, and frequently in tree shrews; no synapses were observed in Syrian hamsters and rats. In the pineal gland of all the animals, GABA-immunoreactive cell bodies were not detected, and sympathetic fibers were not immunoreactive for GABA. These data indicate that GABAergic fibers are main pinealopetal projections from the brain. In view of the difference in the distribution of these fibers, central GABAergic innervation may play a more significant role in nonrodents than in rodents. The frequent occurrence of GABAergic synapses on pinealocytes in the tree shrew suggests that GABA released at these synapses directly controls activity of pinealocytes of this animal.

Light and electron microscopic immunocytochemical study on the innervation of the pineal gland of the tree shrew (Tupaia glis), with special reference to peptidergic synaptic junctions with pinealocytes.

Conventional and immunocytochemical, light- and electron-microscopic studies on the innervation of the pineal gland of the tree shrew (Tupaia glis) were made. Neuropeptide Y (NPY)-immunoreactive fibers, which were abundantly distributed in the gland, disappeared almost completely after superior cervical ganglionectomy, suggesting that these fibers are mostly postganglionic sympathetic fibers. By contrast, tyrosine hydroxylase (TH)-immunoreactive fibers, which were less numerous than NPY-fibers, remained in considerable numbers in ganglionectomized animals, indicating the innervation of TH-positive fibers from extrasympathetic sources. Bundles of substance P (SP)- or calcitonin gene-related peptide (CGRP)-immunoreactive fibers, entering the gland at its distal end, were left intact after ganglionectomy. SP-fibers were numerous, but CGRP-fibers were scarce in the gland. SP-immunoreactive fibers were myelinated and nonmyelinated, and were regarded as peripheral fibers because of the presence of a Schwann cell sheath. NPY- and SP-immunoreactive fibers and endings were mainly localized in the pineal parenchyma. NPY-immunoreactive endings synapsed frequently, and SP-positive ones did less frequently, with the cell bodies of pinealocytes. The results suggest that NPY and SP directly control the activity of pinealocytes. Sections stained for myelin showed that thick and less thick bundles of myelinated fibers entered the gland by way of the habenular and posterior commissures, respectively. Under the electron microscope, the bundles were found to contain also unmyelinated fibers. A considerable number of nerve endings synapsing with the cell bodies of pinealocytes remained in ganglionectomized animals; these endings were not immunoreactive for TH or SP. Such synaptic endings may be the terminals of commissural fibers.

Peptidergic peripheral nervous systems in the mammalian pineal gland.

The distribution and density of tyrosine hydroxylase (TH) and neuropeptide Y (NPY)-immunoreactive, sympathetic fibers and calcitonin gene-related peptide (CGRP)-, substance P (SP)-, and vasoactive intestinal polypeptide (VIP)-immunoreactive, non-sympathetic fibers in the pineal gland, the effects of superior cervical ganglionectomy (SCGX) on these fibers, and the location of their terminals in the pineal gland were compared between rodents and non-rodents. A dense network of TH/NPY-positive fibers is present all over the pineal gland. A less dense network of CGRP/SP- or VIP-positive fibers occurs in the whole pineal gland of non-rodents, but these fibers are usually confined to the superficial pineal gland in rodents. After SCGX, some TH/NPY-fibers remain only in the deep pineal gland in rodents, whereas considerable numbers of these fibers persist throughout the gland in non-rodents. Thus, the remaining fibers, probably originating from the brain, may be more numerous in non-rodents. Since CGRP-, SP- or VIP-immunoreactive fibers in the pineal capsule can be traced to those in the gland, and since these fibers are ensheathed by Schwann cells, it is concluded that these fibers belong to the peripheral nervous system. However, the existence of SP-positive central fibers cannot be denied in some species. In the superficial pineal gland of rodents, sympathetic terminals are mostly localized in perivascular spaces, whereas the parenchymal innervation by sympathetic fibers in the pineal gland is more dense in non-rodents than in rodents. Synapses between sympathetic nerve terminals and pinealocytes occur occasionally in non-rodents, but only rarely in the superficial pineal gland of rodents. The occurrence of the synapses may depend on the frequency of intraparenchymal sympathetic terminals.

Quantitative light microscopic study on the heterogeneity in the superficial pineal gland of the rat.

BACKGROUND: The previous results regarding regional and day-night differences in pinealocyte size in rats are conflicting. The relationships between these differences and the vascularity and sympathetic innervation have scarcely been investigated. METHODS: Wistar-King rats, kept under light/dark 12:12, were killed at midday or midnight in October. The nuclear density of pinealocytes in the superficial pineal was measured on the dorsoperipheral, dorsocentral, ventroperipheral, and ventrocentral regions at distal, middle, and proximal levels at daytime and nighttime. The total area of blood vessels per unit area at daytime and nighttime and total length of tyrosine hydroxylase (TH)-immunoreactive fibers per unit area at daytime were determined on the same regions at the same levels. RESULTS: Pinealocyte size was larger toward the distal levels and in the periphery than in the center at any level. The area of blood vessels and length of TH fibers were also larger toward the distal levels; the former in the ventral region and the latter in the dorsal and ventral regions were larger in the periphery than in the center. Ventral pinealocytes, but not dorsal ones, showed day-night changes in size. Prominent day-night rhythms in area of blood vessels occurred in the ventral region, where TH fibers were more abundant than in the dorsal region. CONCLUSIONS: Pinealocyte size shows the distal to proximal and peripheral to central gradients, which may be related to the differential distribution of blood vessels and sympathetic fibers. Since pinealocytes and blood vessels, showing prominent day-night changes in size, are localized in the more richly innervated regions, sympathetic fibers may play an important role in controlling these rhythms.

[Readmission among discharged psychiatric patients and it's correlates].

OBJECTIVE: This study investigated the incidence of readmission among discharged psychiatric patients and examined factors predicting early readmission. METHODS: A cohort of 343 patients, who had been hospitalized involuntarily to mental hospitals for medical care and custody, and who were discharged between April 1991 and September 1993, in two areas served by Fukuoka Prefectural Yamato Health Center (n = 163) and Tagawa Health Center (n = 180) were followed up until November 1994. RESULTS: The readmission rates within 6 months of discharge in Yamato Health Center and Tagawa Health Center were 17% and 22%, and within 1 year were 30% and 31% respectively. From Cox's proportional hazards model, alcohol/drug abuse, many of previous admission, long length of recent hospitalization, payment of medical care cost from public assistance, complication of physical disorders, living without a person responsible for custody after discharge, no request of health center's service by the hospital and discharge from a large-scale mental hospital were significantly related to increased risk of readmission. Not a few patients could not be followed up completely because of early drop-out of treatment. The drop-out rates within 1 month of discharge in Yamato Health Center and Tagawa Health Center were 10% and 26%, and within 1 year were 15% and 27% respectively. CONCLUSIONS: The rates of readmission and drop-out of treatment among discharged psychiatric patients were considerably high. This study clarified that rapid establishment of a mental health system supporting the mentally disabled in a community is an urgent need in Japan.

Regional differences in the pineal gland of the cotton rat, Sigmodon hispidus: light microscopic, electron microscopic, and immunohistochemical observations.

Light microscopic, electron microscopic and immunohistochemical observations of the various portions of the pineal gland of the cotton rat (Sigmodon hispidus) were made. The volume of the proximal half occupied about 30% of the whole organ, and pinealocytes were slightly smaller in size in the proximal portion than elsewhere. The distal and intermediate portions contained few interstitial cells and numerous astrocytes, but the proximal portion lacked interstitial cells and had more abundant astrocytes than elsewhere. Astrocytes, which were immunoreactive for glial fibrillary acidic protein, mainly lined the pericapillary spaces in the distal and intermediate portions, but in the proximal portion these cells often surrounded isolated or groups of pinealocytes. In the distal and intermediate portions, abundant sympathetic fibers and less numerous non-sympathetic, peptidergic fibers were mainly localized in the pericapillary spaces; these fibers were sparsely distributed in the parenchyma close to interstitial cells or astrocytes. In the proximal portion, non-sympathetic fibers were scarce and sympathetic fibers were distributed abundantly and almost exclusively in the parenchyma. Most of the sympathetic fibers were adjacent to astrocytes and, occasionally, made specialized contact with them. Fenestrae in the capillary endothelium were numerous in the distal portion but absent in the proximal portion. Thus, marked differences in structure existed between the distal and proximal portions of the pineal gland of the cotton rat suggesting that both portions are functionally dissimilar. In addition, the present study indicates that the proximal portion of the cotton rat was well developed and showed morphological features similar to the deeply situated pineal glands of other mammals.

Sprouting of non-sympathetic myelinated and unmyelinated fibres in response to chronic sympathetic denervation in the pineal gland of the Chinese hamster, Cricetulus griseus.

We have examined the effects of chronic sympathetic denervation on non-sympathetic myelinated and unmyelinated fibres in the superficial pineal gland of the Chinese hamster (Cricetulus griseus), using LM, EM and immunohistochemistry. The results suggest that non-sympathetic, myelinated and unmyelinated fibres enter the superficial pineal gland at its distal portion by way of the nervi conarii, and that these fibres are immunoreactive for calcitonin gene-related peptide or substance P. Non-sympathetic, myelinated and unmyelinated fibres in the superficial pineal gland increased in number following chronic superior cervical ganglionectomy. The number of unmyelinated fibres in the nervi conarii also increased in ganglionectomized animals. Thus, the numerical increase of calcitonin gene-related peptide or substance P fibres found in the superficial pineal gland after long-term sympathectomy may be due to sprouting of these fibres. It is speculated that the growth of non-sympathetic, myelinated and unmyelinated fibres and myelination of the former fibres occurring after sympathectomy are caused by nerve growth factor-related mechanisms.

Immunohistochemical studies on sympathetic and non-sympathetic nerve fibers and neuronal cell bodies in the pineal gland of cotton rats, Sigmodon hispidus.

Immunohistochemistry revealed the presence of tyrosine hydroxylase (TH)-, neuropeptide Y (NPY)-, calcitonin gene-related peptide (CGRP)- and substance P (SP)-immunoreactive nerve fibers and SP-immunoreactive neuronal cell bodies in the pineal gland of the cotton rat (Sigmodon hispidus). Abundant TH- and NPY-immunoreactive fibers were distributed evenly throughout the gland; less numerous CGRP- and SP-immunoreactive fibers were distributed in the superficial pineal and the stalk, but were scarce in the deep pineal. All the immunoreactive fibers were usually found around blood vessels. Since TH- and NPY-immunoreactive fibers in various pineal regions disappeared completely with superior cervical ganglionectomy, these fibers are all considered postganglionic sympathetic fibers. Intrapineal CGRP- or SP-immunoreactive fibers decreased considerably in number following superior cervical ganglionectomy, suggesting that some sympathetic fibers contain CGRP or SP. Bilateral bundles of nerve fibers under the transverse sinuses, corresponding to the nervi conarii, contained TH-, NPY-, CGRP- and SP-immunoreactive fibers, which continued into those distributed in the pineal capsule. In the nervi conarii, fibers immunoreactive for TH and NPY disappeared after superior cervical ganglionectomy, but those immunoreactive for CGRP and SP persisted. Thus, non-sympathetic, CGRP- and SP-immunoreactive fibers, together with sympathetic fibers, are presumed to enter the gland by way of the nervi conarii. Neuronal cell bodies, containing SP-like immunoreactivity and being possibly parasympathetic in nature, occurred occasionally in the superficial pineal.

Effect of magnetic field on pineal gland volume and pinealocyte size in the rat.

Light microscopic observations on the superficial pineal gland of Wistar-King rats were made to examine whether or not pineal volume and pinealocyte size, expressed as nuclear density, at daytime or nighttime are affected by long-term exposure to 50 Hz rotating magnetic field (MF) at 5.0 microT. Determinations of pineal volume and pinealocyte size were repeated twice (April and October) during the year. Size of pinealocytes in MF-exposed and sham-exposed rats exhibited, in addition to the difference between peripheral and central regions, regional differences in a proximodistal direction; pinealocytes in the distal and middle-peripheral regions were usually larger than those in the proximal and middle-central regions at daytime or nighttime. In October, distal and proximal pinealocytes showed significant day-night changes in size in sham-exposed rats, but not in MF-exposed animals. The situations in the two groups were almost reversed in April. Significant day-night differences were scarcely found in pinealocyte size in the middle region in the two groups. Throughout the study, pineal volume and pinealocyte size in each region were generally the same between MF-exposed and sham-exposed rats at daytime or nighttime. The results suggest that pinealocytes in the distal and proximal regions, but not those in the middle region, are affected by MF-exposure; day-night differences in sizes of distal and proximal pinealocytes appear in April and disappear in October under the influence of MF. MF may exert an effect on mechanisms controlling day-night rhythms of pinealocyte size in the rat.

Postnatal changes in nerve cells and myelinated fibers in the pineal gland of cotton rats (Sigmodon hispidus). A light microscopic study.

Intrapineal neurons and intracapsular and intrapineal myelinated fibers were studied by light microscopy in male cotton rats (Sigmodon hispidus) at various postnatal ages. Until 300 days of age, neurons are encountered mainly in the distal third of the organ; neuronal nuclei are largest in this region. The remaining regions contain only a few neurons with smaller nuclei. In all regions, nuclear sizes of the neurons increase, and neuronal numbers reduce by half until 100-200 days. Neurons thus consist of different populations of cells, and the aging changes in neuronal nuclei seem to be related to the differentiation of the organ. The disappearance of neurons and the enlargement of the nuclei of remaining neurons proceed simultaneously with age. Intracapsular and intrapineal myelinated fibers appear at 20 days and their density increases with age. As the myelinated fibers are distributed mainly in the distal region and its capsule, these fibers are related topographically to intrapineal neurons. The intracapsular myelinated fibers continue into bilateral bundles of myelinated fibers found under the transverse sinuses, corresponding to the nervi conarii. Since the intracapsular fibers and the fibers traced between the capsule and the parenchyma bifurcate in a proximal direction, the intracapsular fibers are considered to run proximally, some entering the organ.

Effect of photoperiod on pineal gland volume and pinealocyte size in the Chinese hamster, Cricetulus griseus.

Male adult (200-day-old) Chinese hamsters (Cricetulus griseus) raised from weaning under either LD 16:8 or LD 8:16 were used. The pineal gland of the Chinese hamster consists of superficial (major) and deep (minor) components and a continuous, or interrupted, narrow parenchymal stalk interposed between them. The volume of the superficial pineal including the parenchymal stalk is greater under LD 16:8 than under LD 8:16. Under both photoperiods, pinealocytes in the superficial pineal have larger nuclei and more abundant cytoplasm than those in the deep pineal. Nuclei in the superficial pineal appear pale and usually have irregular profiles, whereas those in the deep pineal appear dark and have round profiles. In the superficial pineal, pinealocyte nuclei are larger, paler, and more irregular; and, in addition, nuclear density is lower under LD 16:8 than under LD 8:16. Similar, but less prominent, photoperiod-induced changes occur in the volume of the deep pineal, the size of pinealocytes, and pinealocyte nuclear morphology in the deep pineal. The results indicate that the development and differentiation of pinealocytes in both pineal portions may be advanced under long photoperiods and delayed under short photoperiods, although pinealocytes in the deep pineal may remain not fully differentiated even in adults. Since testicular weights and body weights are similar under both photoperiods, the photoperiod may exert marked influences on the development of the pineal gland without affecting reproductive activity and growth rates of animals.

Twenty-four-hour changes in pinealocytes, capillary endothelial cells and pericapillary and intercellular spaces in the pineal gland of the mouse. Semiquantitative electron-microscopic observations.

Semiquantitative electron-microscopic observations on the pineal gland of dd-mice were carried out to determine whether 24-h rhythms exist in pinealocytes, pericapillary and intercellular spaces and capillary endothelial cells. Nuclear and cytoplasmic areas of pinealocytes and the area of condensed chromatin in pinealocytes showed inversely related circadian rhythms; the former two increased, whereas the latter decreased, during the light period. The extent of pericapillary and wide intercellular spaces exhibited 24-h changes, with an increase and decrease occurring during the light period and the dark period, respectively. The cross-sectional area of endothelial cells decreased and the number of fenestrae increased during the light period; this was reversed during the dark period. The results suggest that the increase in the nuclear and cytoplasmic areas of pinealocytes, the area of pericapillary and wide intercellular spaces and the number of fenestrae, and the decrease in the area of condensed chromatin and endothelial cells during the light period may be related to an increase in synthetic activity of pinealocytes in the mouse.

Comparisons of sizes of pinealocyte nuclei and pinealocytes in young and adult Chinese hamsters (Cricetulus griseus) under different photoperiod conditions.

Semiquantitative electron microscopic observations on pinealocytes of the Chinese hamster (Cricetulus griseus) were made to compare the sizes of pinealocyte nuclei and pinealocytes as a whole, their nuclei and cytoplasm together, over a 24-hr period, between young animals (60-70 days old) and adult animals (120-130 days old) under LD 12:12 and between adults under LD 12:12 and LD 14:10. Under LD 12:12, similar 24-hr rhythms exist in the nuclear area and the area of pinealocytes of young animals, whereas in adults these values exhibit no significant 24-hr rhythm. In adults under LD 14:10, there is no significant 24-hr rhythm in the nuclear area, but the area of pinealocytes shows a statistically significant 24-hr rhythm. Thus, in the Chinese hamster, the relationships between nuclear area and area of pinealocytes with time of day vary, depending on the age of animals as well as different photoperiodic conditions, although they differ only slightly.