A comparative study of the pineal complex in the deep-sea fishes Bathylagus wesethi and Nezumia liolepis

Summary The pineal complexes of two deep-sea fishes, Bathylagus wesethi (family Bathylagidae) and Nezumia liolepis (family Macrouridae), were studied with both light and electron microscopy. Receptor and supportive cells were identified in the pineals of both species. The presence of receptor cells suggests that the pineals function in photoreception. Ganglion cells could be identified only in B. wesethi. A dorsal sac and a paraphysis were found in B. wesethi; both structures are absent in N. liolepis.Several trends were found when the results of this study were compared with those of a study on the pineal complex of another deep-sea fish, the myctophid Triphoturus mexicanus (McNulty and Nafpaktitis, 1976). Two of these trends, which are correlated with the vertical distributions of the species studied, suggest an increase in the photosensitivity of the pineals. These are: 1) an increase in the average number of outer segment lamellar membranes per receptor cell, and 2) an increase in the ratio of receptor cells to nerve fibers in the pineal stalks.A functional relationship between the dorsal sac, paraphysis, and pineal central lumen was suggested. The relationship may involve secretory activities.     

The Structure and Development of the Pineal Complex in the Lanternfish Triphoturus mexicanus (Family Myctophidae)

The pineal complex of the lanternfish Triphoturus mexicanus was studied by light and electron microscopy. Receptor, supportive and ganglion cells were identified in the pineal end-vesicle. The presence of receptor cells and the prominent pineal window strongly suggest a photoreceptive function. Numerous unmyelinated nerve fibers were also found in the end-vesicle. These converge to form part of the pineal stalk. Two kinds of cells (Types I and II) were distinguished in the dorsal sac, which is very well developed. Type I cells are non-ciliated ependymal cells and contain large parcels of glycogen. Type II cells contain large lipidlike inclusions and a distinct band of filaments around the entire periphery of the cell. Receptor cells appear very early in the development of T. mexicanus. They seem to originate from ependymal cells lining the roof of the third ventricle.     

鲫鱼松果体的显微和超微结构研究

本文作者发现鲫鱼的松果体与一般硬骨鱼不同,它除由背囊和背囊内褶中松果管所组成的松果体外,还有退化的旁突体和副松果体.背囊是单层柱状纤毛上皮,其腔与第3脑室相通,松果管由光感觉细胞、支持细胞、节细胞、丰富的血管和无髓神经纤维构成.松果体既是光感受器又有内分泌的功能.       

The morphology and histology of the pineal organ in roach, Rutilus rutilus (L.)

The pineal organ in the roach, Rutilus rutilus (L.), is covered by a semi-transparent area, the pineal window. Beneath this the pineal is attached to a long robust stalk, lying just under the parietal bone. The pineal is attached to the brain through the dorsal sac. Three cell types have been identified histologically. These are the sensory cells, supporting cells and the ganglia cells. The inner segment of the sensory cells respond to PAS and AF staining, while the remaining cells respond to Orange G, LG, or Acid Fuchsin. The evidence suggests that the roach pineal may have a dual photosensory and glandular function.     

Regional distribution and cellular expression of tryptophan hydroxylase messenger RNA in postmortem human brainstem and pineal gland

The characterization and cellular localization of tryptophan hydroxylase mRNA in the human brainstem and pineal gland were investigated by using northern blot analysis and in situ hybridization histochemistry. Northern analysis of human pineal gland revealed the presence of two mRNA species that were absent in RNA isolated from human raphe. In situ hybridization experiments revealed very dense hybridization signal corresponding to tryptophan hydroxylase mRNA in cells throughout the pineal gland. In contrast, tryptophan hydroxylase mRNA was heterogeneously distributed in neurons in the dorsal and median raphe nuclei. Within the dorsal raphe, the ventrolateral and interfascicular subnuclei contained the greatest number of tryptophan hydroxylase mRNA-positive neurons. Also, the cellular concentration of tryptophan hydroxylase mRNA varied widely within the dorsal and median raphe. Comparison of the cellular concentration of tryptophan hydroxylase mRNA between the pineal gland and the raphe nuclei revealed an 11- and 46-fold greater average grain density of tryptophan hydroxylase mRNA positive cells in the pineal gland compared with the dorsal and median raphe, respectively. These findings are the first to demonstrate the cellular localization of tryptophan hydroxylase mRNA in the human brain and pineal gland as well as heterogeneity in the cellular concentration within and between these tissues.     

The pineal complex: a morphological and immunohistochemical comparison between a tropical (Paracheirodon axelrodi) and a subtropical (Aphyocharax anisitsi) characid species

Cardinal neon Paracheirodon axelrodi and bloodfin tetra Aphyocharax anisitsi are two species of characids with high trade value as ornamental fish in South America. Although both species inhabit middle water layers, cardinal neon exhibits a tropical distribution and bloodfin tetra a subtropical one. In this work, we carried out an anatomical, histological and immunohistochemical study of the pineal complex of P. axelrodi and A. anisitsi. In both species, the pineal complex consisted of three components, the pineal and parapineal organs and the dorsal sac (DS). The pineal organ was composed of a short, thin pineal stalk (PS), vertically disposed with respect to the upper surface of the telencephalon, and a pineal vesicle (PV), located at the distal end of the PS and attached to the skull by connective tissue. The pineal window (PW), a site in the skull where the luminal information accesses the pineal organ, appeared just above the latter structures. In the epidermis of P. axelrodi's PW, club cells were identified, but were not observed in the epidermis of A. anisitsi's one. With respect to the DS, it appeared to be folded on itself, and was bigger and more folded in A. anisitsi than in P. axelrodi. Immunohistochemical assays revealed the presence of cone opsin‐like and rod opsin‐like photoreceptor cells in the PS and PV. These results provide a first insight into the morphological assembly of the pineal complex of both species, and contribute to a better understanding of the integration and transduction of light stimuli in characids. J. Morphol. 277:1355–1367, 2016. © 2016 Wiley Periodicals, Inc.     

Morphology of the pineal organ in the salamander Ensatina eschscholtzi

The pineal organ of Ensatina eschscholtzi, a terrestrial and secretive species of salamander of the family Plethodontidae, is a photoreceptive structure lying on the dorsal surface of the diencephalon. The pineal is flattened with a broad lumen and consists of three cell types: photoreceptors, supportive cells, and neurons. Pineal photoreceptors are typical vertebrate photoreceptors and possess outer segment formations which, however, are frequently contorted and disorganized. Sloughing of apical portions of outer segments and vesiculation along the lateral edges of outer segment membrane disks are consistently observed and presumed to represent mechanisms of outer segment membrane recycling. Photoreceptors have basal processes which synapse with neural dendrites. Synapses between photoreceptor basal processes are occasionally observed. All synapses are characterized by synaptic ribbon structures of variable number, size, and configuration. Dense-core vesicles are occasionally observed mingled with clear synaptic vesicles within photoreceptor basal processes. Supportive cells within the pineal function in phagocytosis and recycling of shed outer segment membrane material, and neurons are localized at the lateral margins of the organ. The latter send axons into the ipsilateral side of the dorsal diencephalon. The pineal organ of Ensatina shows marked variation in overall size (cell total), cell type proportions, absolute neuron number, and ratio of photoreceptor number to neuron number for individual pineals. None of these morphological parameters is correlated with body size, sex, or season, and it is assumed that such variability represents significant variation in photosensory capabilities. It is suggested that the pineal organ of Ensatina is a partially degenerate photoreceptive structure.     

Morphological comparison of the ampullae of Lorenzini of three sympatric benthic rays

This study investigated and compared the morphology of the electrosensory system of three species of benthic rays. Neotrygon trigonoides, Hemitrygon fluviorum and Maculabatis toshi inhabit similar habitats within Moreton Bay, Queensland, Australia. Like all elasmobranchs, they possess the ability to detect weak electrical fields using their ampullae of Lorenzini. Macroscopically, the ampullary organs of all three species are aggregated in three bilaterally paired clusters: the mandibular, hyoid and superficial ophthalmic clusters. The hyoid and superficial ophthalmic clusters of ampullae arise from both dorsal and ventral ampullary pores. The dorsal pores are typically larger than the ventral pores in all three species, except for the posterior ventral pores of the hyoid grouping. Ampullary canals arising from the hyoid cluster possessed a quasi‐sinusoidal shape, but otherwise appeared similar to the canals described for other elasmobranchs. Ultrastructure of the ampullae of Lorenzini of the three species was studied using a combination of light, confocal and electron microscopy. All possess ampullae of the alveolar type. In N. trigonoides and M. toshi, each ampullary canal terminates in three to five sensory chambers, each comprising several alveoli lined with receptor and supportive cells and eight to 11 sensory chambers in H. fluviorum. Receptor cells of all three species possess a similar organization to those of other elasmobranchs and were enveloped by large, apically nucleated supportive cells protruding well into the alveolar sacs. The luminally extended chassis of supportive cells protruding dramatically into the ampullary lumen had not previously been documented for any elasmobranch species.     

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.     

Two morphological types of pineal window in catfish in relation to photophase and scotophase activity: a morphological and experimental study

The pineal window is a transparent/translucent pineal covering on the dorsal surface of the cranium of certain fishes and is associated with light reactions of fish. In the present study, catfish species Clarias batrachus, Heteropneustes fossilis, Mystus vittatus, M. seenghala, and M. cavassius were examined for the type of pineal window present. Two morphologically different types of pineal window were found: an opaque-looking pineal window in C. batrachus and H. fossilis and a translucent type of pineal window in M. vittatus, M. seenghala, and M. cavassius. The distributional pattern of pigments in the melanophores at the pineal window were studied in terms of Melanophore Index (MI). In all of the species studied, a pineal foramen, a subepidermal lens-like tissue, and pineal end vesicle were present. Experiments were carried out on catfish having the opaque pineal window, as it is uncommon in catfish. Catfish with normal and shielded pineal window were exposed to conditions of artificial constant illumination (LL) and darkness (DD) to evaluate the effects of altered photoperiods on the state of pigmentation of melanophores at the pineal window. Recordings of diel activity patterns, which are light dependent in catfish, were carried out under both natural and artificial photoperiods in fish with a normal or shielded window in order to assess its functional nature. The existence of two morphologically and functionally different types of pineal window in a relatively closely related group of catfish has been demonstrated in this study. The nature of the opaque type of pineal window has been reconsidered based on new experimental evidence.     

Light and electron microscopic studies on the pineal tract of rainbow trout, Salmo gairdneri.

The pineal tract of rainbow trout from the pineal end vesicle to the posterior commissure was studied by light and electron microscopy. Five types of nerve fibres (photoreceptor basal process, ganglion cell dendrite, electron-lucent fibre and synaptic vesicles, myelinated and unmyelinated axons) and two modes of synapses (photoreceptor basal process ganglion cell dendrite and axon terminal with synaptic vesicles-photoreceptor basal process synapses) are distinguishable in the proximal region of end vesicle. The two distinct synaptic associations with the photoreceptor basal process suggest two different (excitatory and inhibitory) control of pineal sensory activity. At the distal portion of stalk about two thousand nerve fibres converge into dorsal and ventral bundles. Posterior to the habenular commissure several small branches run out laterally from the ventral bundles to the basal margin of the ependyma, but not into the habenular commissure. The dorsal bundle passes through the dorsal side of the subcommissural organ and runs ventral to the posterior commissure. The pineal tract is composed of unmyelinated axons, electron-lucent nerve fibres and myelinated axons. The number of fibres increases throughout the stalk and reaches the maximum number at the opening of pineal lumen to IIIrd ventricle, however, the number of fibres then decreases through the subcommissural organ and posterior commissure. This increase and decrease of nerve fibres suggest the continuous participation of axonal fibres of pineal nerve cells and the ramification or branching of pineal tract, respectively.     

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     

Glutamate receptor subunit delta2 is highly expressed in a novel population of glial-like cells in rat pineal glands in culture

The mammalian pineal gland uses L-glutamate as an intercellular chemical transmitter to regulate negatively melatonin synthesis. To receive glutamate signals, pinealocytes express at least three kinds of glutamate receptors: metabotropic receptor types 3 and 5 and an ionotropic receptor, GluR1. In this study, we examined whether or not the fourth class of ionotropic receptor, delta, which is known for its nondefinitive molecular function and its unique expression pattern in brain, is expressed in pineal gland. RT-PCR analyses with specific probes indicated the expression of mRNA of delta2 but not that of delta1 in pineal gland and cultured pineal cells. Western blotting analysis with polyclonal antibodies specific to the carboxyl-terminal region of the delta2 receptor recognized a single 110-kDa polypeptide of cerebellar membranes and specifically immunostained Purkinje cells. The delta2 antibodies recognized a 110-kDa polypeptide of pineal membranes and specifically immunostained huge glial-like cells with the occasional presence of several long, branching processes in a pineal cell culture. delta2 is not uniformly distributed throughout the cells and is relatively abundant at the periphery of the cell bodies and long processes, where the terminals of synaptophysin-positive processes of pinealocytes, a site for glutamate secretion, are frequently present. The delta2-positive cells constitute a very minor population among total pineal cells (approximately 0.03%). Double immunolabeling with delta2 antibodies and antibodies against marker proteins for pineal interstitial cells clearly distinguishes delta2-positive pineal cells and other known interstitial cells, including glial fibrillary acidic protein- or vimentin-positive glial-like cells. These results indicated that the delta2 glutamate receptor is expressed in a novel subpopulation of pineal glial-like cells in culture and suggest the presence of a glutamate-mediated intercellular signal transduction mechanism between pinealocytes and delta2-expressing cells. The pineal cells may provide a good experimental system for studies on the function of glutamate receptor delta2.     

Ultrastructure of the rectal sac, the site of water vapour uptake from the atmosphere in larvae of the oriental rat flea Xenopsylla cheopis

A portion of the hindgut of Xenopsylla cheopis larvae, the rectal sac, is extraordinarily enlarged and differentiated into a specialized organ displaying well-elaborated transporting epithelia. This organ is unique among insect recta because of gutter-like arrangement of two ultrastructurally distinct cell types with an asymmetrical position of the membranemitochondria complexes. The cells of the dorsal gutter have highly folded apical plasma membranes on the lumen side. The cells of the ventral gutter show deep infoldings of the basal plasma membrane on the haemolymph side. Less elaborate folds originate from the opposite sides of the cells. The nuclei of the dorsal epithelial cells are located in the basal part; those of the ventral cells in the apical portion of the cell. The ultrastructural features of the rectal sac, its ventilation synchronized with rhythmic opening of the anus during phases of active water vapour uptake, as well as the reversible arrest of uptake by experimental occlusion of the anus, reveal that condensation and uptake of atmospheric water take place in this organ.     

Pineal corpora arenacea produced by arachnoid cells in the bat Myotis blythi oxygnathus

There are corpora arenacea among the cell layers of the arachnoid on the dorsal surface of the pineal organ of the bat (Myotis blythi oxygnathus). The pineal arachnoid consists of electron lucent cells connected by cell injunctions to flat sheets and sandwiched on both sides by electron-dense cell rows. Among the superficial cell layers, collagen fibrils form loose bundles. In the electron-lucent cells, pinocytotic vesicles, rough surfaced endoplasmic reticulum, active Golgi areas and granular vesicles of various sizes can be found. Electron dense cells display fewer cytoplasmic organelles than the light ones. Lying between and below the hemispheres and cerebellum the pineal arachnoid does not contact the dura mater directly, therefore it continues on its both sides into arachnoid trabeculae. Corpora arenacea occur in lacunar enlargements of the arachnoid, first of all in the thickened dorsal portion of the pineal leptomeninx. The acervuli are insulated by collagen fibrils and exhibit concentric layers of various density. Needle-shaped structures resembling hydroxyapatite crystals were found in these concentric layers. There was no sign of formation of acervuli in the pinealocytes or elsewhere in the pineal nervous tissue proper. These findings confirm that view that corpora arenacea can be produced by the pineal arachnoid. The formation of acervuli is accompanied by secretory and resorptive phenomena of arachnoid cells.     

Modification of the deep pineal after ablation of the superficial epiphysis in the golden hamster]

In various species (among Rodents: Mesocricetus auratus, Cricetus cricetus, Mus musculus, Meriones libycus, M. shawi et M. crassus; among Cheiroptera: Artibeus jamaicensis etc), the pineal complex is formed by two different portions one superficial situated under the cerebral dura and one deep portion situated between the habenular and posterior commissures. Superficial and deep portions are separated by a tractus of fibers and some pineal cells. Superficial epiphysectomy in the Mesocricetus auratus determines an increase in volume of the deep pineal. The nuclear diameter is also enlarged in this case. The functions of both superficial and deep pineal seem to be closely related. Superficial epiphysectomy is not followed by involution or hypoactivity of the deep pineal. Superficial epiphysectomy also determines changes in the subcommissural and subfornical organs.     

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     

The effect of alphaMSH on beta-adrenergic receptor mechanisms in the rat pineal.

The effects of αMSH on beta adrenergic receptor response of rat pineal cells were studied in vitro. Responses measured were membrane hyperpolarization, measured with a micro-electrode, and cyclic AMP formation, measured by radio immunoassay. Normal resting membrane potential of pineal cells is approximately -40mV. Addition of NE produces a dose-dependent hyperpolarization of these cells. The addition of αMSH in vitro produces a very slight, but significant, depolarization, and markedly attenuates subsequent NE responses. αMSH has no effect on the cyclic AMP content of pineal glands, but again attenuates the action of NE in producing increased cellular cyclic AMP. These results suggest that αMSH may modulate pineal responsiveness.     

Substance P-like immunoreactivity in the parietal eye visual system of the lizard Uta stansburiana

Summary We examined the parietal eye visual system of the iguanid lizard Uta stansburiana for the presence of substance P-like immunoreactivity by use of both immunofluorescence and peroxidase-antiperoxidase techniques. In the parietal eye no substance P-containing somata were found; however, its plexiform layer contained small (ca. 1 m diam) immunoreactive fibers. These fibers apparently originate outside the parietal eye. Immunoreactive fibers also were found in the parietal nerve, the dorsal sac, and the leptomeninx of the pineal gland. No labeled somata were observed in any of these regions in either normal or colchicine treated animals. Previously we demonstrated that a system of centrifugal fibers to the parietal eye originates from neurons in the dorsal sac (Engbretson et al. 1981). The apparent absence of substance P-containing neurons in the dorsal sac suggests that the substance P-containing fibers in the parietal eye are not the previously observed centrifugal fibers. The source of the substance P-containing fibers in the parietal eye is unknown. The pars dorsolateralis of the left medial habenular nucleus receives a dense substance P-positive projection. No such projection was seen in the right habenula. Simultaneous visualization of the terminals of ganglion cells of the parietal eye (labeled with orthograde intraaxonally transported horseradish peroxidase) and substance P-like immunofluorescence showed that the locus of habenular immunoreactivity is distinct from the projection field of the parietal eye. Thus the substance P-positive terminals in the habenula do not originate in the parietal eye. Transection of the parietal nerve confirmed this conclusion.