Neuronal degeneration in the pineal ganglion during the post-hatching development of the domestic fowl

Summary The frequency of pineal ganglia associated with the pineal tract, and the numbers of acetylcholinesterasepositive neurons in these ganglia were studied in the domestic fowl during the post-hatching period by means of the acetylcholinesterase method. Furthermore, the degeneration of nerve cells in pineal ganglia of 40-day-old domestic fowl was investigated in detail at the electron-microscopic level. The rate of pineal organs containing one or more ganglia was 50% in 2- to 13-day-old, 38% in 40-day-old, and only 10% in 1-year-old domestic fowl. In parallel, the number of acetylcholinesterase-reactive nerve cells that constitute individual pineal ganglia decreased after hatching. Various degrees of neuronal degeneration were found in the pineal ganglia: swelling of the endoplasmic reticulum, electron-dense degeneration of the cytoplasm, and pyknosis of the nerve cell nucleus. Clusters of macrophages containing numerous lysosomes filled with debris-like material were scattered in the ganglion. In addition, plasma cells were observed in association with degenerating nerve cells. These results confirm the suggestion that the loss of acetylcholinesterase-positive nerve cells in the pineal ganglia of the domestic fowl is due to naturally occurring, programmed neuronal cell death. This process is discussed with reference to phenomena of cell death observed in other components of central nervous system.Fellow of the Alexander von Humboldt Foundation, Bonn, Federal Republic of GermanyThe authors are indebted to Professor A. Oksche and Dr. H.-W. Korf (Giessen) for stimulating discussions     

A pineal ganglion associated with the pineal tract in the domestic fowl

Summary A ganglion-like aggregate consisting of acetyl-cholinesterase-positive neurons was demonstrated in the pineal organ of the domestic fowl by means of light and electron microscopy. This ganglion is located in juxtaposition with the pineal tract at the posterior (caudal) aspect of the pineal stalk. Numerous large and small neurons formed the ganglion in 40-day-old domestic fowl. Some of these nerve cells established direct neuro-neuronal contacts, others were surrounded by satellite cells. These ganglion cells displayed axo-somatic and axo-dendritic synapses. The above-mentioned cluster of nerve cells may be considered as a pineal ganglion. Its central or peripheral nature is open to discussion. Send offprint requests to: Dr. K. Wake, Department of Anatomy, Faculty of Medicine, Tokyo Medical and Dental University, Yushima, Bunkyo-ku, 113, Japan     

Separation of the pineal vesicle from the wall of the third ventricle during the post-hatching development of the chicken

Summary According to light- and electron-microscopic observations the pineal organ of the 3-day-old chicken consists of a prominent end vesicle and a tapering parenchymal stalk. During this stage the pineal lumen is in open communication with the third ventricle. However, in the 40-day-old chicken, which still possesses a well-developed end vesicle, the proximal portion of the pineal stalk displays regressive changes leading to local fragmentation. At this stage the pineal stalk is reduced, and the pineal lumen is missing. In 1-year-old chickens the parenchyma of the proximal portion of the stalk is further diminished, and in 3-year-old domestic fowl is completely displaced by bundles of collagenous fibers, only some nerve fibers being present. This post-hatching pineal development may reflect the sequence of changes leading from pineal sense organs to pineal glands.This work was supported by a grant-in-aid for Scientific Research from the Ministry of Education, Science and Culture of Japan     

Regressive post-hatching development of acetylcholinesterase-positive neurons in the pineal organs of Coturnix coturnix japonica and Gallus gallus

Summary Distribution and number of acetylcholinesterase-positive neurons were studied in the Japanese quail and the domestic fowl during the post-hatching period by means of the acetylcholinesterase method. For comparison, the development of the catecholamine-containing (sympathetic) pinealopetal fibers of the domestic fowl was demonstrated with the use of the glyoxylic acid method. The number of acetylcholinesterase-positive ganglion cells in the pineal organs of both avian species decreased rapidly after hatching, with a concentration of these elements in the basal portion (stalk) of the pineal organ.In 3-day-old chickens, perivascular catecholamine-containing nerve fibers penetrate the antero-lateral walls of the pineal organ and are found exclusively in the interfollicular and perivascular tissues. In 13-day-old and adult fowl, these fibers increase in number and terminate not only in the interfollicular space but also in the neuroepithelial parenchyma of the pineal body.The ontogenetic regression of the sensory structures paralleled by an expanding sympathetic innervation in the pineal organ of a galliform species resembles somewhat the process of phylogenetic transformation leading from pineal sense organs to pineal glands.This work was supported by a grant (No. 56480080) from the Ministry of Education, Science and Culture of Japan.Fellow of the Alexander von Humboldt Foundation (1982).     

The preoptic area of the domestic fowl

Summary The preoptic area of the domestic fowl (Gallus gallus) was studied by means of the Golgi technique. At least two regions can be recognized: (i) a medial and (ii) a lateral area, clearly distinguishable laterally from the adjacent telencephalic regions. The dendritic organization of the preoptic area is quite uniform. The neurons can be classified as isodendritic elements. The magnocellular elements are few and irregularly scattered mostly in the periventricular grey of the medial preoptic area. Of relevant interest is also the observation of some bipolar and horizontal neurons in the dorsal part of the medial preoptic area, near the anterior commissure.     

Fine structural studies on the thyroid gland of the normal domestic fowl

Summary The ultrastructure of the thyroid gland of the domestic fowl has been investigated and found to be similar to that of mammals. The differences were found at subcellular level in the distribution of the dark bodies which were mainly apical and in the sizes of primary lysosomes. These were found to range from 100 to 500 nm in diameter. All organelles described in mammals as being concerned with the production of thyroglobulin and the two hormones thyroxine and triiodothyronine were found to be present.     

Localization of corticotropin-releasing factor-containing neurons in the brain of the domestic fowl

Summary The corticotropin-releasing factor (CRF)-containing neurons were investigated in the brain of the domestic fowl by means of the peroxidase-antiperoxidase technique at the light-microscopic level. The detection of CRF-immunoreactivity was facilitated by silver intensification. CRF-containing perikarya were found in the paraventricular, preoptic and mammillary nuclei of the hypothalamus and in some extrahypothalamic areas (nuclei dorsomedialis and dorsolateralis thalami, nucleus accumbens septi, lobus parolfactorius, periaqueductal gray of the mesencephalon, nucleus oculomotorius ventralis). Immunoreactive nerve fibers and terminals were demonstrated in the external zone of the median eminence and the organum vasculosum of the lamina terminalis. These results indicate that an immunologically demonstrable CRF-neurosecretory system also exists in the avian central nervous system.     

The hypothalamic magnocellular system in the domestic fowl

Summary In the rostral hypothalamus of the domestic fowl, the magnocellular neurosecretory nuclei show a peculiar differentiation. Golgi studies of the supraoptic and paraventricular nuclei of the fowl reveal at least two major cell types: 1) large multipolar neurons, and 2) small interneurons. Golgi impregnations provide a detailed cytoarchitectural picture of the large-sized cells; the latter may well correspond to the neurosecretory cells demonstrated in the same regions by selective staining, and immunocytochemical and electron microscopical techniques.Electron microscopically, neuronal perikarya are observed to contain variable amounts of neurosecretory granules (100–200 nm in diameter; mean diameter of 160 nm) scattered throughout the cytoplasm. The diameters of these granules do not differ statistically in the two principal nuclear areas examined. The perikarya of these neurons display only a few axosomatic synapses containing electron-lucent and dense-cored vesicles (70–90 nm in diameter). Numerous nerve terminals of this type also end on the dendritic ramifications in the surrounding neuropil.     

Electron microscopy of the rabbit pineal organ in vitro

The results presented in this study show that the rabbit pineal organ cultured in vitro retained its in vivo fine structure for at least eight days. However, the Golgi complex in the light pinealocytes stopped forming dense core vesicles while vesicle-crowned ribbons increased in number. After addition of norepinephrine to the culture medium, the Golgi complex once more began the production of dense core vesicles. Terminals of light pinealocytic processes then often contained Golgi dense core vesicles in close contact with the cell membrane, suggesting the release of the vesicular content into the intercellular and perivascular spaces. A close topographical relationship between Golgi dense core vesicles and vesicle-crowned ribbons was observed.     

Ultracytochemistry of the synaptic ribbons in the rat pineal organ

Summary The synaptic complexes of the rat pinealocytes are neither cholinergic nor adrenergic. In the synaptic vesicles, a neurotransmitter carrier substance of lipid nature reacting with OsO₄-Zn I₂ mixture (similar to that present in both cholinergic and adrenergic vesicles) was not found.In addition, there were no indications of glucose-6-phosphatase or thiamine-pyrophosphatase activity in the synaptic vesicles. Thus, it appears that the synaptic vesicles do not originate from the rough or smooth endoplasmic reticulum.The synaptic ribbons do not contain carbohydrates, are of protein nature and possess some chemical resemblance to microtubules and microtubular bouquets.Appropriate ultracytochemical reactions have not shown detectable quantities of sodium and calcium ions in pinealocyte synaptic complexes.Grateful acknowledgment is made to Mr. P.-A. Milliquet for technical assistance and to Dr. T. Jalanti (C.M.E., Lausanne) for his help in the use of the X-ray microanalyser.Dedicated to Professor Dr. med. G. Töndury on the occasion of his 70th birthday.     

Post-natal development of the pineal organ in the hamsters Phodopus sungorus and Mesocricetus auratus

Summary In Phodopus sungorus, as in other mammals, the pineal organ forms an important link in the transduction of photoperiodic information to the endocrine system. The sympathetic innervation, via the superior cervical ganglion, controls the metabolism of serotonin and melatonin in the pineal, which in turn is involved in the control of the gonads. In the present study, the post-natal development of this system was investigated. Specimens 1, 5, 10, and 15 days post partum (p.p.) and adults were treated with monoamine-oxidase-inhibitor and perfused under ether anesthesia via the aorta with a buffer containing glyoxylic acid, formaldehyde and Mg⁺⁺. The brains were then dissected out and treated according to Falck-Hillarp for fluorescence microscopy and microspectrofluorometry. Day 1: The nervi conarii had reached the pineal capsule, but only in a few cases was the pineal organ invaded and then only by a few fibers. Day 5: A rich green-fluorescing net of fibers was present in the entire organ, stalk and lamina intercalaris. No 5-HT fluorescence was observable. Day 10: Similar to the stage at 5 days a rich green-fluorescing nerve fiber net was observed throughout the pineal and a yellow fluorescence in the pineal perikarya. Day 15: The general appearance resembles the adult. The nerve fibers are masked by the intense yellow fluorescence of the pineal perikarya. Fading of the latter, however, allows the catecholamine fluorescence to be seen. Golden hamsters at an age of 15 days p.p. show a similar appearance to Phodopus at an age of 15 days. Microspectrofluorometric determinations indicated the catecholamine to be noradrenaline, and confirmed a 5-HT/5-HTP origin of the yellow fluorescence appearing between day 5 and day 10. The amount of 5-HT/5-HTP was considerably less at day 10 than at day 15 or in adults. Sympathectomy by extirpation of the superior cervical ganglion abolished the catecholamine fluorescence completely in the pineal body, stalk and lamina intercalaris.Supported by grants from the Swedish Natural Science Research Council (to P. Meurling and Th. van Veen), and the Royal Physiographic Society of Lund     

Immunocytochemical demonstration of met-enkephalin in the central nervous system of the domestic fowl

Summary In the central nervous system of the male domestic fowl, met-enkephalin (ENK) immunoreactive perikarya and fiber tracts as well as extensive but sharply delimited fiber networks were visualized by means of the PAP technique. The most striking results were: (1) The demonstration of an association of ENK-containing structures with branchial nerves; (2) the spatial relationship of ENK-containing perikarya and fibers to somatostatin (SOM) and arginine-vasotocin (AVT)-immunoreactive systems; (3) the presence of dense and extensive ENK fiber networks within (a) the caudo-basal wall of the third ventricle and (b) the septal-preoptic area; in both regions mainly ENK fibers, but also SOM and AVT fibers, may cross to the contralateral side.     

Ultrastructural observations on synaptic ribbons in the pineal organ of the goldfish

Summary Synaptic ribbons in the pineal organ of the goldfish were examined electron microscopically with particular attention to their topography. These structures were formed of parallel membranes, which were poorly preserved with OsO₄ fixation and could be extracted from thin sections with pronase indicating their proteinaceous nature. Synaptic ribbons were closely apposed to the plasma membrane bordering dendrites of ganglion cells, but were also related to processes of both photoreceptor and supportive cells. Their close proximity to invaginations of the plasma membrane and portions of the endoplasmic reticulum suggest that they are involved in the turnover of cytoplasmic membranes. Tubular and spherical organelles of unknown function are also described.     

Evidence for a nervous connection between the brain and the pineal organ in the guinea pig

Summary Following the injection of horseradish peroxidase into the pineal organ of the guinea pig, approximately 30 nerve fibers were demonstrated in the pineal stalk due to the retrograde transport of the tracer enzyme in these elements. Finely branched extensions of these nerve fibers are directed toward the distal portion of the pineal organ. This projection of central nervous elements enters the pineal organ via the habenular or posterior commissures. Neuronal perikarya projecting into the pineal organ are found in the region of the paraventricular nucleus near the border of the third ventricle.In partial fulfillment of the requirements for the degree of Dr. med., Faculty of Medicine, Justus Liebig University of Giessen     

Scanning electron microscopic analysis of the linings of the fourth ventricle in the domestic fowl

Summary Surface features of the ependymal linings of the fourth ventricle in the fowl were analyzed employing the scanning electron microscope (SEM). On the floor of the median sulcus, each ependymal cell has a solitary cilium, whereas on both sides of the sulcus, cilia are so densely distributed that the details of the underlying cell surface are usually obscured. On the roof of the fourth ventricle, except for the surface of the ciliated groove where numerous cilia are present, the ependymal cells are polygonal in shape, and the center of each cell possesses an aggregate of ten to twenty cilia. Cell surfaces of the choroid tela are entirely covered with delicate microvilli and possess clumped cilia. The ependymal cell surfaces of the area postrema are dome-like in shape. Each ependymal cell has a solitary cilium and shows a smooth surface free of microvilli.This work was supported by a Scientific Research Grant, No. 144017, from the Ministry of Education of Japan to Professor M. Yasuda     

The fine structure of the vesicular component of the ultimobranchial gland of the domestic fowl

Summary The ultrastructure of the polymorphic vesicular component of the ultimobranchial gland of the domestic fowl (Gallus gallus domesticus) has been described in detail, together with the structure of the cell strands interconnecting the vesicles and the parathyroid nodules lying within the ultimobranchial stroma. The vesicles frequently appear to arise from the nodules by way of the cell strands. The strands show a structure of their component cells intermediate between that of the parathyroid and the vesicular cells, although the position at which the strand changes from an essentially parathyroid structure to an essentially vesicular structure is very variable. The degree and kind of secretory activity within different cell types has been described. A review of the structure of ultimobranchial glands throughout the vertebrates shows that similar tissue with a similar secretory potential has been observed in all vertebrate classes, suggesting a functional significance for this part of the gland.     

Vasopressin- and oxytocin-containing fibres in the pineal gland and subcommissural organ of the rat

Summary Vasopressin and oxytocin were specifically demonstrated in the rat brain using the unlabelled antibody-enzyme method and purification of the first antiserum. Vasopressin and oxytocin fibres extend via the subcommissural organ or habenular commissure into the pineal stalk and terminate in the anterior part of the pineal organ. In addition, immediately adjacent to the subsommissural organ many vasopressin-containing fibres run caudally toward the central grey. These results are discussed in relation to the proposed presence of vasotocin in the pineal gland.This study was supported by the Foundation for Medical Research, FUNGOThe authors wish to thank Dr. D.F. Swaab and Prof. J. Ariëns Kappers for their suggestions and critical remarks