Immunocytochemical localization of S-100 protein in the hypophysis and saccus vasculosus of the elasmobranchs Mustelus manazo and Scyliorhinus torazame

Summary S-100 protein-immunoreactive cells were demonstrated by immunocytochemical procedures in the hypophysis and saccus vasculosus of two species of elasmobranchs (Mustelus manazo and Scyliorhinus torazame). In the saccus vasculosus of M. manazo, immunoreactivity was detectable exclusively in the fibrous portions interposed between the epithelial layer and the blood vessels. In the neurohypophysis, tanycytes and astrocytes of the median eminence were immunostained, but only a few labeled cells were found in the neurointermediate lobe. In S. torazame, the neurohypophysis displayed a similar distribution of immunoreactivity, but there were no labeled cells in the saccus vasculosus. In both species, none of the glandular cells of the hypophysis displayed immunoreactivity. Electron-microscopic examination showed that the immunostained cells in the saccus vasculosus correspond to astrocytes.     

Changes in the Saccus Vasculosus Epithelium During the Smoltification Process in Atlantic Salmon (Salmo salar)

The changes that take place in the saccus vasculosus epithelium during smoltification of Atlantic salmon (Salmo salar) are described. The major cell type of the epithelium, the coronet cells, have a number (mean = 32) of spherical protrusions connected with a non-motile cilium to the apical part of the cell. The protrusions extend into the saccus lumen. In parr the protrusions become more spherical during the spring season, while in smolt the opposite process takes place. Parr coronet cells have most of the mitochondria in the apical part of the cell, while in smolt they are concentrated basally. The relative areal fraction of heterochromatin in the nuclei increase from 23 to 29 during the smoltification process. The results are consistent with the view that the coronet cells take an active part in the smoltification process.     

Secretion by ependymal cells of the neurohypophysis and saccus vasculosus of Polypterus ornatipinnis (Osteichthyes)

Electron microscopic studies of the neurohypophysis and saccus vasculosus of the bichir (Polypterus ornatipinnis) reveal an apocrine release of secretory material by ependymal cells (e.g., crown cells). The secretory material appears to migrate along the microtubular apparatus and the ciliary filaments to the cell buds. It is postulated that the formation of the buds and their release is mediated by cilial action. Secretion buds are noted in the cerebrospinal fluid and vascular sinusoids. Bulbous projections of neurohypophyseal secretory tracts extend into the ependymal lumen which also contains elementary secretion granules. Specialized “liquorkontaktneurone” are interspersed with ependymal cells. An interrelation between the hypothalamus, neurohypophysis, saccus vasculosus, and the meta-adenohypophysis is postulated.     

The saccus vasculosus of Anguilla anguilla (L.) from larva to adult

Summary The ultrastructure of coronet cells of the saccus vasculosus has been studied in specimens of Anguilla anguilla (L.) at different stages of its life cycle. At all the stages observed coronet cells are composed of a basal and an apical part, the latter bearing globules with primary vesicles. In the larva (a marine form) and in the fully metamorphosed small eel at the time of entry into freshwater the narrow lumen and the vesicles within the apical globules are filled with electron-dense material. In forms in which adaptation to freshwater has occurred, the saccus lumen appears expanded, the apical globules are better developed, and the electron-dense material has disappeared. It is suggested that the two situations observed represent different functional states of the organ, in relation to different conditions of environmental salinity.The authors gratefully acknowledge the contribution of Dr. G. Andreoli, of the University of Parma, who provided the Atlantic larvae for this study.     

Possible Ascidian Counterpart to the Vertebrate Saccus Vasculosus with Reference to Pyura tessellata (Forbes) and Boltenia echinata (L.)

The morphology of the hitherto unknown larva of Pyura tessellata (Forbes) is described and the discovery of an auxiliary brain vesicle is presented. The vesicle was also found in another pyurid species Boltenia echinata (L.). The auxiliary vesicle communicates with the sensory vesicle at the level of the statocyte and is lined with cubodial epithelial cells carrying a 2 μm globular structure projecting into the lumen. These cells very closely resemble primitive tunicate coronet cells previously reported and coronet cells typical for the saccus vasculosus found in elasmobranchs and many ganoids and teleosts.     

Light microscopic and ultrastructural study of the development of the saccus vasculosus in the rainbow trout,Oncorhynchus mykiss

This study using light and electron microscopy indicates that the saccus vasculosus is distinguishable in 9-mm embryos and grows continuously throughout embryonic development to the adult stage. In the saccus vasculosus, epithelial mitoses are observed in all stages studied. Phases of centriologenesis, ciliogenesis, and globule formation have been characterized in developing coronet cells. During the phase of centriologenesis, new centrioles appear in association with pre-existing centrioles and not on deuterosomes. After ciliogenesis, each cilium differentiates to a globule almost at the same time as the other cilia of the coronet cell. The inner membrane system of the globules seems to derive from the ciliary plasma membrane. This membrane system often produces membrane whorls during the development. The different phases of coronet cell development have been found in the same individual and in all the stages studied except the 9-mm embryo. Cerebrospinal fluid-contacting neurons are observed in the saccus epithelium from the 12-mm embryos on and are distinguishable from coronet cells in their early formative stages. The three cell types of the saccus vasculosus increase continuously in number during development. Nerve processes are found in the saccus vasculosus of embryos, whereas differentiated synapses appear later in the fry. The significance of continued coronet cell formation is discussed in relation to a putative coronet cell and/or a globule renewal cycle in the adult.     

The neuronal system of the saccus vasculosus of trout (Salmo trutta fario and Oncorhynchus mykiss): an immunocytochemical and nerve tracing study

The neuronal system of the saccus vasculosus of two species of trout was studied with immunocytochemical methods and carboindocyanine-dye (DiI) tract-tracing. The cerebrospinal-fluid-contacting neurons of the saccus were immunoreactive for gamma-aminobutyric acid (GABA), glutamic acid decarboxylase (GAD), and neuropeptide Y (NPY). Immunostaining of alternate sections of the saccus vasculosus of fry with anti-GAD and anti-NPY indicated that these substances were colocalized. The tractus sacci vasculosi and the neuropil of the nucleus sacci vasculosi were also immunoreactive to these substances. The GABA, GAD, and neuropeptide Y immunoreactivity of the saccus vasculosus system appeared early in trout ontogeny. After applying DiI to various levels of the tractus sacci vasculosi of adult trout, we observed massive bilateral saccular projections to the nucleus sacci vasculosi and could follow the course of the sacco-thalamic tract. This tract extended in the subependymal region of the thalamus rostral to the nucleus sacci vasculosi and split into two small tracts that reached the subhabenular-preoptic region. Sacco-thalamic fibers formed extensive periependymal plexuses along their trajectory. Interestingly, no clear evidence of the existence of a saccopetal system was obtained. On the basis of these results, we postulate that the saccus vasculosus system modulates the function of centers of the posterior tubercle and periventricular thalamus. Received: 5 August 1996 / Accepted: 4 January 1997     

The Structure of the Hypothalamohypophysial Portal Vascular System in Acipenser ruthenus L. (Chondrostei)

The vascularization of the pituitary region in Acipenser ruthenus L. (Chondrostei) is described. The adenohypophysis has no direct arterial supply but is fed exclusively by a pituitary portal system supplied through a pair of infundibular arteries. Distinct portal vessels connect the lateral part of the primary plexus of the neurohaemal area (the median eminence) with the secondary plexus of the pituitary gland. The primary plexus enters the pars distalis paramedially, apparently without the formation of distinct portal vessels. The neuro-intermediate lobe receives its blood supply exclusively from the primary plexus. The plexus intermedius gives off capillaries to the parenchyma of the intermediate lobe (an intermediate lobe sinus system). The saccus vasculosus receives (1) a “direct” supply, i.e. branches originating directly from the cerebral arteries and (2) an “indirect” supply, i.e. capillaries from the primary plexus. The pars distalis is drained into an unpaired ventral hypophysial vein, while a dorsal hypophysial vein, also unpaired, drains the plexus intermedius. These two veins join to form the unpaired hypophysial vein. The findings are discussed from comparative and functional viewpoints.     

Über Aktivitätsphasen im Epithel des Saccus vasculosus von Etmopterus spinax

Summary The coronet cells in the saccus vasculosus of Etmopterus spinax L. were investigated histologically and histochemically. Inclusions of granula in the coronet cells were observed in 8 specimens. The granules react positivly as lipid and protein containing substances. They are attached to the Einschlußkörper in the cell base and accumulate around the nuclei or in the inner fibre apparatus of the cell apex. The diameter of the granules reaches 2,5 in the cell base and diminishes to less than 1 in the apical structures. These granula are interpreted as a sign of secretion into the third ventricle.This secretory activity is high in gravid females (31,4–54,7 % of the cells) and low in juvenile females or specimens in which no ovulation could be stated (0–4,7 %). Gravid females show differences in the distribution pattern of the granula within the cells, thus indicating a phasic activity.The coronet cells in the saccus vasculosus of Etmopterus seem to be involved in the secretory phenomena related to gravidity.

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Die Untersuchungen wurden mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft durchgeführt.     

Histophysiology of the saccus vasculosus of the Indian freshwater goby Glossogobius giuris Ham. (Teleostei)

The author studied the structure and functions of the saccus vasculosus of the Indian freshwater goby Glossogobius giuris (Ham.). The saccus is ovoid, is localized on the ventral surface of the brain and is lodged between the inferior lobes. It consist of several loculi lined with coronet cells and is bathed with blood from surrounding sinusoids. The coronet cells are variably shaped and have a conspicous central nucleus. It is suggested that the purpose of the saccus vasculosus is to act as a storage site for carbohydrates to the brain. By converting glycogen to acid mucopolysaccharides, the coronet cells are involved in glycogen metabolism.     

The development of a saccus vasculosus in Tilapia mossambica Peters and other cichlids

Observations are made on the development and the presence of a Saccus Vasculosus in Tilapia mossambica and other cichlids.
In those cichlids that do not possess a saccus vasculosus the development of this organ is apparently suppressed in a very early stage of life.
A saccus vasculosus is present in all those American and Indian cichlids that were investigated, but this organ was not found in cichlids indigenous to the African continent.     

Localization by immunofluorescence of a gastrin-like substance in the brain of the rainbow trout,Salmo gairdneri

Summary With the aid of an indirect immunofluorescence technique neurones containing a gastrin-like substance were identified in the brain of Salmo gairdneri. The perikarya of these neurones appear to be located along the periventricular part of the nucleus lateralis tuberis between the hypophysial stalk and the most rostral tip of the saccus vasculosus. The fibres of these perikarya run rostrally toward the hypophysis, where they can be followed in the protrusions of the neurohypophysis into the proximal pars distalis. Here the bundle of immunoreactive fibres divides into numerous smaller bundles and into single fibres. Immunohistochemical specificity tests have shown this immunoreactive substance to belong to the gastrin group, sharing an antigenic determinant with cholecystokinin (CCK) and pentagastrin (common aminoacid sequence Trp-Met-Asp-Phe). A possible function of these gastrin (or CCK)-containing neurones in the rainbow trout is discussed.     

On the presence and localization of glycogen accumulations inside coronet cells of the saccus vasculosus of the dogfish (Scylliorhinus caniculus)

Summary In several coronet cells of the saccus vasculosus of Scylliorhinus large quantities of glycogen occur, as shown by light and electron microscopy. The significance of glycogen as an energy storage necessary for a transcellular ion transport process taking place in the coronet cells is discussed.The authors thank Dr. F.C.G. van de Veerdonk, W. F. Jansen and W. F. G. Flight for reading the manuscipt and for their critical remarks. They are also indebted to Mr. H. van Kooten and his staff for their valuable photographic assistance.     

Ultrastructure of sensory receptors in ascidian tadpoles

Summary An analysis of fine structure and function was conducted on three types of receptors in the cerebral vesicle of two species of ascidian tadpoles (Ciona intestinalis andDistaplia occidentalis). Theocellus is composed of one pigmented, cup-shaped supportive cell, an estimated 15–20 sensory cells, and three lens cells, each with a large body of granules (glycogen ?). The outer segments of the photoreceptoral processes are modified cilia, one per sensory cell, consisting of many lamellae, formed by infoldings of the ciliary membranes, and axonemes of 9 + 0 doublets of microtubules. The lamellae are homologous to retinal disks of vertebrates.Hydrostatic pressure (?)receptors are modified cilia containing tubules which open to the lumen of the cerebral vesicle. These receptors closely resemble the globular, ciliary processes of coronet cells in the saccus vasculosus of fishes. Thestatocyte is a onecell gravity receptor. The part extending into the lumen of the brain contains the nucleus and a large black body which is thought to function as a float. The foot-piece of the cell is firmly anchored in the brain wall; the neck is probably the site of generation of signals. This study was supported by grant 10292 from the USPHS. The senior author held an appointment in the Miller Institute for Basic Research in Science, University of California, during part of the investigation. We appreciate the assistance of Emily Reid, artist; Jean L. Brandenburger, research associate; Professor Gerald Westheimer, consultant; Charles Spowart, collector; and the UCLA Brain Information Service, and the hospitality of Dr. Robert L. Fernald, Director, Friday Harbor Laboratories, University of Washington, U.S.A.     

Relationships between the tubular system in the globules of the coronet cells of the saccus vasculosus and the cerebrospinal fluid in Gasterosteus aculeatus form leiurus (Teleostei)

Summary In the stickleback, Gasterosteus aculeatus, the globules of the coronet cells in the saccus vasculosus contain a tubular system that most probably communicates permanently with the lumen of the saccus. Only very few openings were found in random ultrathin sections. Injecting peroxidase into the cerebrospinal fluid revealed the communication between this tubular system and the cerebrospinal fluid. As early as 1 h after peroxidase injection the tracer was detected in the tubular system. This system increases the potential exchange surface between the coronet cells and the content of the saccular lumen, and may also facilitate the access of components of the cerebrospinal fluid (CSF), even of high molecular weight, into the globules. It remains to be determined whether the intratubular condensations (granules) are, as often believed, of secretory origin or are formed by accumulation of components of the CSF.With the technical collaboration of Melle A. Meunier     

Metabolic activity in the saccus vasculosus of the rainbow trout,salmo gairdneri (richardson)

Summary Labelling experiments to test mucopolysaccharide synthesis in the saccus vasculosus of the rainbow trout gave negative results and provoked analysis of succinic dehydrogenase activity in the coronet cells. Such activity was almost absent in the fresh-water living rainbow trout but was intense in sea water adapted individuals as well as in a sea water teleost, the flounder. Conversely, in sea water adapted rainbow trout which had been transferred back to fresh-water, activity had again decreased. In the sea water adapted rainbow trout high succinic dehydrogenase activity was paralleled by strongly positive staining with alcian blue, indicating the presence of mucopolysaccharides. The latter fishes also showed incorporation of N-acetyl-³H-glucosamine. Our findings indicate a role for saccus vasculosus in adaptation for osmoregulation in sea water and favour the hypothesis of a secretory function for the coronet cells.This work has been supported by grants from the Royal Physiographical Society of Lund and the Faculty of Natural Sciences at the University of Lund. We are greatly indebted to Mrs. Lena Eriksson, Mrs. Kristine Fogelström and Mrs. Lena Svenre for excellent technical assistance.     

Effects of stress on carbohydrate metabolism in the teleost Notopterus notopterus (Pallas)*

The effects of different types of stre35 on carbohydrate metabolism in N. notopterus were investigated. Starvation alone brings about a signifiant increse in the glycogen content of the saccus vasculosus and a significant decrease in the brain glycogen concentration. The increased glycogen concentration in the saccus vasculosus may be a device to safeguard the brain against glucose deficiency during starvation stress. Rapid depletion of the muscle glycogen following fasting shows that the muscle glycogen IS the readily utilizable source of energy during starvation. Exposure of N. noropierus to air brought about an increaSe in the liver glycogen and blood glucose levels but did not affect other paramcters studied. Physical exhaustion of N. notopierus is noticed within 1–2 min of exercise. The readily available source of energy for exercise is the muscle glycogen. and the lactic acid produced is probably metabolized in the muscle itself. Saccus vasculosus glycogen. though inde-pendent of changes in the blood glucose levels, may possibly be controlled by variations in the brain glycogen.     

The hypothalamo-hypophysial vascular relationship in Indian fresh-water goby, Glossogobius giuris (Ham.).920u.

The hypothalamo-hypophysial vascular relationship and intra-hypophysial vasculatisation have been described in order to understand the regulatory mechanism of hypothalamic control over the functions of the pituitary gland. In Glossogobius giuris, the disposition of the blood vessels in the head region is on typical teleostean pattern with certain modifications. The nucleus preopticus is supplied through the nucleus preopticus artery, a small blood vessel arising from the anterior branch of the posterior cerebral artery, whereas the pituitary gland receives blood through a pair of hypophysial arteries. The blood from the pituitary is drained off by the pituitary veins whch pour their blood into the supra-orbital sinus. The anterior cerebral vein after taking the blood from anterior part of the brain including the hypothalamus and the nucleus preopticus joins with the supra-orbital sinus. The hypothalamo-hypophysial portal system is absent in this fish. The saccus vasculosus receives blood from the posterior cerebral artery through a small blood vessel and is collected by a prominent saccus vasculosus vein which pours blood into the supra-orbital sinus before it joins the infra-orbital sinus to form the heat vein. There seems to be no physological connection between the saccus vasculosus and pituitary gland. The highly vascularised neurohypophysis interdigitate with the pars intermedia and extends upto the proximal pars distalis. The blood vessels are restricted to the neurohypophysial extensions only. However, in the rostral pars distalis the blood vessels are present but the neurohypophysis does not extend to this part. The blood capillaries enter the rostral pars distalis from the capillary network on the surface of pituitary gland along with the connected tissue covering of the pituitary. The neurohypophysis shows a greater vascularisation in comparison to that of the other glandular part of the pituitary gland. In the present study of Glossogobius giuris, though an extensive ramification of neurohypophysis occurs with the pars intermedia and the proximal pars distalis, the neurosecretory axons do not innervate the endocrine cells of the pituitary gland and the blood vessels are found restricted to the neurohypophysial extensions except that of the rostral pars distalis. The neuro-vascular way of hypothalamic control over the functions of the pituitary gland seems to be justified as the neurosecretory fibres have been found associated with the blood vessels.