Electron microscopic and experimental studies of the pineal organ in the white-crowned sparrow,Zonotrichia leucophrys gambelii

Summary The structure of the pineal organ of Zonotrichia leucophrys gambelii, as revealed by light- and electron-microscopy, resembles that of Passer domesticus (Oksche and Kirschstein, 1969; Ueck, 1970). The typical cellular element is the pinealocyte with certain basic structural features of the pineal photoreceptors of lower vertebrates (see Oksche, 1971). However, instead of the characteristic, cone-like outer segments, there are, as in other species of birds, only bulbous cilia with ectopic whorls of lamellae. This structure of the outer segment is, in a sense, contrary to the demonstration of synaptoid contacts, numerous unmyelinated, and occasional myelinated nerve fibers by electron microscopy. In Nissl preparations it was possible to demonstrate typical nerve cells. The pinealocytes of Z. l. gambelii are secretory; their Golgi complex forms granulated vesicles (800–1,400 Å in diameter) that belong to the group of granular inclusions characteristic of monoamines. Autonomie nerve fibers course within the connective tissue capsule of the pineal organ. In many pinealocytes of Z. l. gambelii, the granular endoplasmic reticulum contains extensively expanded cisternae that are filled with a flocculent material and closely associated with bundles of filaments. In a number of cases such loop-like structures are selectively stainable with aldehyde fuchsin. It was not possible to demonstrate specific secretory activity in the supporting cells. Extirpation of the pineal organ in Z. l. gambelii had no definitely detectable influence on the photoperiodic control of testicular growth.Aves, Passeriformes, Fringillidae.Supported by grants from the Deutsche Forschungsgemeinschaft to Professor Oksche and by the National Science Foundation (GB 11905) to Professor Farner. A part of this investigation was effected while Professor Kobayashi held a Visiting Professorship at the University of Giessen.     

Das visuelle System von Zonotrichia leucophrys gambelii

Zusammenfassung Der Verlauf der Sehbahn und die Lokalisation der optischen Zentren wurden bei Zonotrichia leucophrys gambelii (nordamerikanischer Ammernfink) nach einseitiger Augenexstirpation mit den Techniken von Nauta-Fink-Heimer, Bodian und Bielschowsky erforscht. Die Untersuchungen erstreckten sich über einen Zeitraum von 3 bis zu 120 Tagen nach der Operation. Zonotrichia leucophrys gambelii besitzt ein für Vögel typisches visuelles System. Die Hauptmasse der Optikusfasern endet im Stratum griseum et fibrosum superficiale des Tectum opticum. Weitere zentrale Endgebiete sind: Nucleus geniculatus lateralis, Nucleus lateralis anterior, Nucleus superficialis synencephali, Nucleus externus, tectales Grau und Nucleus ectomamillaris als Kern der basalen optischen Wurzel. Alle Fasern werden im Chiasma opticum total gekreuzt, auch der Tractus isthmo-opticus, ein efferentes Bündel, dessen Ursprung im Nucleus isthmo-opticus zu finden ist. Dieses efferente Fasersystem läßt sich im Stumpf des durchtrennten N. opticus noch 120 Tage nach der Operation gut versilbern. Eine direkte Verbindung von Retina und Hypothalamus war lichtmikroskopisch nicht nachweisbar. Neurosekretorisch aktive Zellen des Hypothalamus können zwar einen engen räumlichen Kontakt mit den optischen Fasern haben, Synapsen sind aber an diesen Stellen nicht zu erkennen. Es werden passagere Opticusfasern beschrieben, die auf dem Weg zum Nucleus lateralis anterior und Nucleus superficialis synencephali den Hypothalamus durchsetzen.

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Neurohistological and experimental studies of the visual system in Zonotrichia leucophrys gambelii

Summary The course of the optic pathways and the positions of the optic centers have been investigated with unilaterally enucleated white-crowned sparrows, Zonotrichia leucophrys gambelii, using the techniques of Nauta-Fink-Heimer, Bodian, and Bielachowsky. The investigation involved birds examined 3–120 days after enucleation. The white-crowned sparrow has a typically avian visual system. The major bundles of optic fibers terminate in the stratum griseum et fibrosum superficiale of the tectum opticum. Further terminal areas are the nucleus geniculatus lateralis, nucleus lateralis anterior, nucleus superficialis synencephali, nucleus externus, the tectal gray, and the nucleus ectomamillaris of the basal optic root. There is a complete crossing of all fibers in the chiasma, including those of the tractus isthmo-opticus, an efferent bundle with its origin in the nucleus isthmo-opticus. This efferent fiber system can be well impregnated in the stump of the sectioned optic nerve up to 120 days after the operation. No direct connection between the retina and hypothalamus could be demonstrated by light microscopy. Hypothalamic neurosecretory cells can occur in close contact with optic fibers but no synapses have been recognized. Some optic fibers pass through the hypothalamus enroute to the nucleus lateralis anterior and the nucleus superficialis synencephali.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft. Herrn Prof. Dr. D.S. Farner, Department of Zoology, University of Washington, Seattle, Wash., danke ich für die Förderung dieser Studien (National Institutes of Health Research Grant No. 5 ROI NB 06187 to Professor D. S. Farner).     

The fine structure of the hypothalamic secretory neurons of the White-crowned Sparrow,Zonotrichia leucophrys gambelii (Passeriformes: Fringillidae)

Summary The fine structures of the neurons and neuropils of the magnocellular supraoptic nucleus and the parvocellular nuclei of the rostral hypothalamus, including the suprachiasmatic and medial, lateral and periventricular preoptic nuclei, and the neuronal apparatus of the organum vasculosum laminae terminalis, have been examined in the male White-crowned Sparrow, Zonotrichia leucophrys gambelii, by correlated light and electron microscopy.The magnocellular supraoptic nucleus is characterized by large neurosecretory perikarya which contain a well developed Golgi complex and densecored granules 1,500–2,200 Å in diameter. The neuropil displays axons, dendrites and glial fibers. Some axonal profiles contain dense-cored vesicles 800–1,000 Å in diameter and clear vesicles 500 Å in diameter. Axo-somatic and axo-dendritic synapses are conspicuous in this nuclear region.The suprachiasmatic nucleus is characterized by an accumulation of small neurons with moderately developed cellular organelles and some dense-cored granules, approximately 1,000 Å in diameter. The profiles of axons within the neuropil contain dense-cored granules 800–1,000 Å in diameter and clear vesicles 500 Å in diameter.The neurons of the medial preoptic nucleus are relatively large and exhibit well developed cellular organelles and dense-cored granules 1,300 to 1,500 Å in diameter. Granular materials are formed within the Golgi complex. The medial preoptic nucleus is rich in secretory perikarya.Occasionally, neurons with granules 1,500–2,200 Å in diameter are encountered in the lateral preoptic and periventricular preoptic nuclei. They may be considered as scattered elements of the magnocellular (supraoptic and paraventricular) system.The organum vasculosum laminae terminalis consists of three layers, i.e., ependymal, internal and external zones, and exhibits a vascular arrangement similar to that of the median eminence. The perikarya of the parvocellular neurons and their axons in the internal zone contain numerous secretory granules ranging from 1,300 to 1,500 Å in diameter.This investigation was supported by Grant No. 5R040 Japan-U.S. Cooperative Science Program of the Japan Society for the Promotion of Science to Professor H. Kobayashi and Professor S.-I. Mikami, by a Scientific Research Grant No. 56019 from the Ministry of Education of Japan to S.-I. Mikami, by support from the Deutsche Forschungsgemeinschaft (Schwerpunktprogramm Biologie der Zeitmessung) to Prof. A. Oksche and by Grant No. GF 33334, U.S.-Japan Cooperative Science Program of the National Science Foundation to Prof. D.S. Farner.Herrn Professor Dr. Dres h.c. Wolfgang Bargmann zu seinem 70. Geburtstag am 27. Januar 1976 gewidmet.     

Twenty-one novel microsatellite DNA loci isolated from the Puget Sound white-crowned sparrow, Zonotrichia leucophrys pugetensis

The white‐crowned sparrow, Zonotrichia leucophrys, has served as a model species for studies of song and reproductive physiology. Here, we describe primers for 21 novel microsatellite loci isolated from the Puget Sound subspecies, Zonotrichia leucophrys pugetensis, which will be useful for parentage and population genetic analyses. Based on genotypes from seven to 22 adult birds from one population, the average number of alleles per locus was 10.9 (four to 21 alleles) and observed heterozygosity varied from 0.50 to 1.00. All loci also amplified products in at least one of three other passerine species tested.     

Fine structure of the vessels of the hypophysial portal system of the White-crowned Sparrow,Zonotrichia leucophrys gambelii

Summary The angioarchitecture of the hypophysial portal system of the White-crowned Sparrow, Zonotrichia leucophrys gambelii, was investigated by electron microscopy in conjunction with light microscopy of serial thick sections.The small arteries or arterioles supplying the primary capillary plexus of the median eminence have the typical form of arterioles.The vessels of the primary capillary plexus, on the surface of the median eminence, with their many fenestrations and pinocytotic vesicles, are typical of the form of capillary usually found in other endocrine organs.The portal vessels in the pars tuberalis have wide perivascular spaces between the basement membrane of the endothelium and that of parenchymal lobules of the pars tuberalis. These perivascular spaces are occupied usually by the perivascular cells, but sometimes contain erythrocytes.The endothelial cells of the portal vessels often protrude into vascular lumen giving the appearance of valve-like structures. These may have a role in the regulation of blood flow.The endothelial cells of the portal vessels are invested by a definitive basement membrane and by the cytoplasm of pericytes which are oriented spirally to the longitudinal axes of the vessels. The pericytes may have a function in the mechanical support of the vascular wall and a contractile function that might regulate the flow rate of blood.The investigation reported herein was supported by a scientific research grant (No. 291049) from the Ministry of Education of Japan to Prof. Mikami; by a grant from the Deutsche Forschungsgemeinschaft to Prof. Oksche; by a grant (5 ROI-NB 06817) from the National Institutes of Health to Prof. Farner, and by a research grant (5 ROI-HE 07240 NEUA) from the National Institutes of Health to Prof. Vitums.     

Plasma corticosterone increases during migratory restlessness in the captive white-crowned sparrow Zonotrichia leucophrys gambelli

Plasma corticosterone increases during the period of spring migration in a variety of bird species. Long-distance migrants show elevations in corticosterone specifically in association with the stage of flight, suggesting that corticosterone may support flight-related processes, for example, locomotor activity and/or energy mobilization. The pattern of corticosterone secretion as it relates to migratory flight has hitherto not been clearly described in migrants that frequently interrupt flight to refuel, for example, the Gambel's white-crowned sparrow (Zonotrichia leucophrys gambelii). The Gambel's white-crowned sparrow fuels by day and expresses peak migratory activity during the first few hours of night. To determine if plasma corticosterone increases in association with the stage of migratory flight also in this short-bout migrant, we induced captive white-crowned sparrows to enter into the migratory condition by placing photosensitive birds on long days (16L:8D) and then evaluated birds for plasma corticosterone and locomotor activity during four time points of the day. Patterns found in long-day birds were compared to those observed in short-day controls (8L:16D). Differences in energy metabolism as determined from plasma metabolites were also evaluated. We found that locomotor activity and corticosterone were significantly elevated at the onset of the dark period, but only in long-day birds. Plasma beta-hydroxybutyrate (a ketone body) was also elevated. Thus, findings suggest that plasma corticosterone and ketogenesis increase in association with migratory restlessness in a short-bout migrant. In fact, corticosterone may play a regulatory role, because it shows a trend to increase already before night-time activity.     

The ACTH-immunoreactive system in the brain of the white-crowned sparrow,Zonotrichia leucophrys gambelii (Passeriformes,Emberizidae)

Summary The ACTH-immunoreactive (ir) system of the avian brain is particularly conspicuous in the male white-crowned sparrow (Zonotrichia leucophrys gambelii). The irperikaryal population is concentrated mainly within the tuberal region, projecting primarily in a dorsal direction: (i) into the striatum; (ii) into rostral diencephalic, septal, hyperstriatal, and thalamic areas; and (iii) into dorsal and ventral areas of the brain stem. Ir-fibers seemingly contact local non-immunoreactive neurons mainly in the accumbens nucleus, septum, dorsal thalamic nuclei, infundibular and interpeduncular nuclei, and in the rostral diencephalon. Neurohemal zones are not supplied by ACTH-ir terminals. Immunocytochemical problems arising from the complexity of the proopiomelanocortin molecule and its derived peptide components are discussed in relation to phylogenetically directed studies, and contradictory results.Dedicated to Professor Dr. T.H. Schiebler on the occasion of his 65th birthday     

The ACTH-immunoreactive system in the brain of the white-crowned sparrow, Zonotrichia leucophrys gambelii (Passeriformes, Emberizidae)

The ACTH-immunoreactive (ir) system of the avian brain is particularly conspicuous in the male white-crowned sparrow (Zonotrichia leucophrys gambelii). The irperikaryal population is concentrated mainly within the tuberal region, projecting primarily in a dorsal direction: (i) into the striatum; (ii) into rostral diencephalic, septal, hyperstriatal, and thalamic areas; and (iii) into dorsal and ventral areas of the brain stem. Ir-fibers seemingly contact local non-immunoreactive neurons mainly in the accumbens nucleus, septum, dorsal thalamic nuclei, infundibular and interpeduncular nuclei, and in the rostral diencephalon. Neurohemal zones are not supplied by ACTH-ir terminals. Immunocytochemical problems arising from the complexity of the proopiomelanocortin molecule and its derived peptide components are discussed in relation to phylogenetically directed studies, and contradictory results.     

The ultrastructure of the cells of Leydig in the white-crowned sparrow (Zonotrichia leucophrys gambelii) in relation to plasma levels of luteinizing hormone and testosterone

In male White-crowned Sparrows subjected to 20 h daily photoperiods there is an approximately 3-fold increase in the plasma concentration of luteinizing hormone (LH) on the first long day after which a quasi-stable level is maintained for at least 42 days. This increase is followed by an increase in numbers of cells of Leydig and an enhancement of their steroidogenic features, a decrease in transitional interstitial cells, and an increase in plasma level of testosterone. With the decline in plasma LH, as photorefractoriness develops, the steroidogenic features of the cells of Leydig undergo disorganization. For as yet unexplainable reasons the plasma levels of testosterone decline before the decrease in plasma LH and before the degeneration of the steroidogenic features of the cells of Leydig.     

Opsin-immunoreactive outer segments and acetylcholinesterase-positive neurons in the pineal complex of Phoxinus phoxinus (Teleostei,Cyprinidae)

Summary The pineal complex of the teleost, Phoxinus phoxinus L., was studied light-microscopically by the use of the indirect immunocytochemical antiopsin reaction and the histochemical acetylcholinersterase (AChE) method.Opsin-immunoreactive outer segments of photoreceptor cells were demonstrated in large numbers in all divisions of the pineal end-vesicle and in the pineal stalk. Moreover, they were found in the roof of the third ventricle, adjacent to the orifice of the pineal recess as well as scattered in the parapineal organ. These immunocytochemical observations provide direct evidence of the presence of an opsin associated with a photopigment in the photosensory cells of the pineal and parapineal organs of Phoxinus. By means of the AChE reaction (Karnovsky and Roots 1964) inner segments of pineal photoreceptors, intrinsic nerve cells, several intrapineal bundles of nerve fibers, and a prominent pineal tract were specifically marked. The pineal neurons can be divided into two types: one is located near the pineal lumen, the other near the basal lamina. The latter perikarya bear stained processes directed toward the photoreceptor layer. A rostral aggregation of two types of AChE-positive nerve cells occurs in the ventral wall of the pineal end-vesicle. The main portion of the AChE-positive pineal tract, which lies within the dorsal wall of the pineal stalk, can be followed to the posterior commissure where some of the nerve fibers course laterally. A few AChE-positive pineal nerve fibers run toward the lateral habenular nucleus via the habenular commissure. In the region of the subcommissural organ single AChE-positive neurons accompany the pineal tract. The nerve cells of the parapineal organ exhibit a moderate AChE activity.These findings extend the structural basis for the remarkable light-dependent activity of the pineal organ of Phoxinus phoxinus. To the memory of Professor Karl von Frisch, pioneer and master in the field of photoneuroendocrine systemsThis investigation was supported by grants from the Deutsche Forschungsgemeinschaft to A.O. (Ok 1/24; 1/25: Mechanismen biologischer Uhren) and to H.-W. K. (Ko 758/1; 758–2)On leave from the 2nd Department of Anatomy, SOTE, Budapest, Hungary     

Validation of an enzyme immunoassay for measurement of excreted estrogen and testosterone metabolites in the white-crowned sparrow (Zonotrichia leucophrys oriantha)

Over the past few years there has been a growing interest in hormonal analyses, despite the cost and difficulty involved in measuring hormones using radioimmunoassays. Therefore, there is a need for alternate procedures, such as enzyme immunoassays, capable of providing similar data to that provided by radioactive methods. Enzyme immunoassays are a less expensive method of measuring estrone conjugates and testosterone. With further work, the enzyme immunoassay also provides the potential to adapt the enzyme assay for use in the field. The data in this study demonstrate that an enzyme assay can accurately detect both endogenous and exogenous hormonal changes in the droppings of white-crowned sparrows. © 1995 Wiley-Liss, Inc.     

Postnatal appearance of luteinizing hormone-releasing hormone (LHRH)-like material in the pineal gland of the rat

Summary Immunoreactive luteinizing hormone-releasing hormone (LHRH)-like material has been demonstrated in the pineal gland of the adult rat. The objective of the present study was to examine the ontogenetic development of this LHRH-like substance in the rat pineal with the peroxidase-antiperoxidase (PAP) method of Sternberger. LHRH-like immunoreactive material was not observed in pineal glands of newborn rats. The amount of material increased progressively from the 6th–12th day of postnatal development. On day 12, the amount of LHRH-like immunoreactivity was consistent and comparable in all pineal glands of male and female animals examined.Supported by NIH Grant 1 R01 HD-12956     

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.     

Pattern of synaptic connections in the pineal organ of the ayu,Plecoglossus altivelis (Teleostei)

Summary Synaptic connections were studied by means of electron microscopy in the sensory pineal organ of the ayu, Plecoglossus altivelis, a highly photosensitive teleost species. Three types of specific contacts were observed in the pineal end-vesicle: 1) symmetrically organized gap junctions between the basal processes of adjacent photoreceptor cells; 2) sensory synapses endowed with synaptic ribbons, formed by basal processes of photoreceptor cells and dendrites of pineal neurons; 3) conventional synapses between pineal neurons, containing both clear and dense-core vesicles at the presynaptic site. Based on these findings, the following interpretations are given: (i) The gap junctions may be involved in an enhancement of electric communication and signal encoding between pineal photoreceptor cells. (ii) The sensory synapses transmit photic signals from the photoreceptor cells to pineal nerve cells. (iii) The conventional synapses are assumed to be involved in a lateral interaction and/or summation of information in the sensory pineal organ. A concept of synaptic relationships among the sensory and neuronal elements in the pineal organ of the ayu is presented.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany     

Immunocytochemical and electron-microscopic investigations of the pineal organ in adult agamid lizards,Uromastix hardwicki

Summary Lacertilian species display a remarkable diversity in the organization of the neural apparatus of their pineal organ (epiphysis cerebri). The occurrence of immunoreactive S-antigen and opsin was investigated in the retina and pineal organ of adult lizards, Uromastix hardwicki. In this species, numerous retinal photoreceptors displayed S-antigen-like immunoreactivity, whereas only very few pinealocytes were labeled. Immunoreactive opsin was found neither in retinal photoreceptors nor in pinealocytes. Electron microscopy showed that all pinealocytes of Uromastix hardwicki resemble modified pineal photoreceptors. A peculiar observation is the existence of a previously undescribed membrane system in the inner segments of these cells. It is evidently derived from the rough endoplasmic reticulum but consists of smooth membranes. The modified pineal photoreceptor cells of Uromastix hardwicki were never seen to establish synaptic contacts with somata or dendrites of intrapineal neurons, which are extremely rare. Vesiclecrowned ribbons are prominent in the basal processes of the receptor cells, facing the basal lamina or establishing receptor-receptor and receptor-interstitial type synaptoid contacts. Dense-core granules (60–250 nm in diameter) speak in favor of a secretory activity of the pinealocytes. Attention is drawn to the existence of receptor-receptor and receptor-interstitial cell contacts indicating intramural cellular relationships that deserve further study.Supported by the Deutsche Forschungsgemeinschaft (Ko 758/31) and the Deutscher Akademischer Austauschdienst (Senior DAAD Research Fellowship to M.A.H.)     

Characteristic pattern of monoaminergic nerve fibers in the pineal organ of the monkey,Macaca fuscata

Summary Monoaminergic nerve fibers were studied in the pineal organ of the monkey, Macaca fuscata, by use of fluorescence and immunohistochemical procedures. Abundant formations of noradrenergic nerve fibers were observed in the pineal organ. They entered the parenchyma in the form of several coarse bundles via the capsule in the distal portion of the organ and spread throughout the organ after branching into smaller units. The density of the autonomic innervation decreased gradually toward the proximal portion of the organ. In the distal portion, numerous nerve fibers formed perivascular plexuses around the blood vessels and some fibers ran as bundles unrelated to the blood vessels in the stroma. Fine varicose fibers and bundles derived from these plexuses penetrated among the pinealocytes. However, only a few intraparenchymal fluorescent fibers were detected in the proximal third of the gland. With the use of serotonin antiserum serotonin-immunoreactive nerve fibers were clearly restricted to the ventroproximal part of the pineal organ. Although the somata of the pinealocytes showed intense immunoreactivity, their processes were not stained. In one exceptional case, clusters of pinealocytes displaying very intense immunoreactivity were found in an area extending from the distal margin of the ventral portion of the pineal stalk to the proximal portion of the pineal organ proper; these cells were bipolar or multipolar and endowed with well-stained processes.     

Ultrastructure of serotonin-containing cells in the pineal organ of Lampetra planeri (Petromyzontidae)

Summary The ultrastructure of the cells containing residual bodies (Collin, 1969) was investigated in the pineal organ of Lampetra planeri. These cells are characterized by their indoleamine metabolism (Meiniel, 1978; Meiniel and Hartwig, 1980). Morphologically, they belong mainly to two types: (1) a photoreceptor cell type, and (2) a pinealocyte cell type. The first type is present in the pineal sensory epithelium and in the atrium, while the second is observed in the deep part of the atrium. Intermediate cell types are rare. All these cells are characterized by the presence of voluminous dense bodies, the 5-HT-storing structures, in their cytoplasm.The elongated cone-type photoreceptor cells show a segmental organization and well-developed outer segments consisting of short disks (2–3 m), while their basal pedicles form synapses with the dendritic processes of neurons. The pinealocytes are spherical or oval in shape, their receptor poles being regressed to cilia of the 9+0 type. In these cells, no synaptic ribbons have to date been observed. In both cell types a Golgi apparatus is present producing dense granules 130 nm in diameter and a polymorphous dense material.The photoreceptor cells most probably respond to light and transmit a sensory (i.e., nervous) message. In addition, they produce and metabolize indoleamines, probably including, melatonin (Meiniel, 1978; Meiniel and Hartwig, 1980). The pinealocytes, in spite of their loss of direct photosensitivity, retain their capacity to metabolize indoleamines (Meiniel, 1978; Meiniel and Hartwig, 1980).The presence, in the same pineal organ, of another photoreceptor cell type (cf. Collin, 1969–1971) differing morphologically as well as biochemically (no detectable indoleamine metabolism) from the photoreceptor cell type described in the present investigation, points to the existence of two different sensory cell lines: (1) a pure photoreceptor line, and (2) a photoneuroendocrine line. The phylogenetic evolution of these two cell lines is discussed in terms of functional analogy.     

Vascular permeability (problem of the blood-brain barrier) in the pineal organ of the rainbow trout,Salmo gairdneri

Summary The problem of the blood-brain barrier in the pineal organ of the rainbow trout, Salmo gairdneri, was investigated following intraperitoneal or intracardial injections of several tracers and dyes with different molecular weights. As demonstrated at the light-microscopic level, repeated injections of trypan blue or horseradish peroxidase (HRP) resulted in an accumulation of these substances in the pineal epithelium (parenchyma). By use of the electron microscope, HRP was found in electron-dense bodies, probably lysosomes, in (i) the endothelial cells and perivascular macrophages 4 h after intraperitoneal injection, (ii) the supporting cells and intrapineal or luminal macrophages 8 h after injection, and (iii) the receptor cells 24 h after injection of the tracer. Ferritin particles penetrated the fenestrated endothelium of pineal capillaries. They were confined to vesicles, vacuoles and the smooth endoplasmic reticulum of the supporting cells as well as to the synaptic vesicles and the smooth endoplasmic reticulum of the pineal photoreceptors. The intercellular passage of tannic acid mixed with the fixative was blocked at the luminal junctional complex separating the pineal lumen from the basal portion of the pineal epithelium. The passive intercellular transport of substances with high molecular weight from the bloodstream to the cerebrospinal-fluid compartment is thus prevented. However, no blood-brain barrier exists for exogenously administered proteins, which are rapidly taken up by pineal cells and actively transported in a transcellular manner.The findings on the blood-brain barrier of the pineal organ of the rainbow trout are discussed with particular reference to the endocrine capacity of pineal sensory organs.Fellow of the Alexander von Humboldt Foundation, Federal Republic of Germany.