Zur Charakterisierung neuronaler und gliöser Elemente im Epithel des Saccus vasculosus von Knochenfischen

Zusammenfassung Das Epithel des Saccus vasculosus des Flußbarsches Perca fluviatilis besteht aus Krönchenzellen, bipolaren Liquorkontaktneuronen (Zahlenverhältnis etwa 41) und Stützzellen. Im Bereich des Saccus kommen Macrophagen vor. Die Krönchenzellen wurden unter verschiedenen Fixierungsbedingungen untersucht. Die Globuli enthalten schlauchförmige Zisternen, die nicht mit den Zisternen des Zellapex in Verbindung stehen. Im Zytoplasma des Zellapex und der Globuli wird bei langdauernder OsO₄-Imprägnation Osmium gebunden. Die Krönchenzellen werden basal von Ausläufern der Stützzellen unterlagert. Sie werden nicht innerviert und entsenden keine Axone.Die bipolaren Neurone sind durch einen im Liquor endigenden Dendriten und ein Axon gekennzeichnet, das in eines der Faserbündel des Epithels eintritt. Der Dendrit trägt 1 bis 2 Zilien. Die Zelle ist reich an Vesikeln und kann am Perikaryon wie an den Ausläufern Synapsen tragen. Im extrazellulären Raum um die Neurone und in vesikulären Strukturen des Apex wird Acetylcholinesterase nachgewiesen.Der Nervus sacci vasculosi dürfte nur afferente Axone von Liquorkontaktneuronen und efferente Fasern, die diese innervieren, enthalten.

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Neuronal and glial cell elements within the epithelium of the Saccus vasculosus in teleosts

Summary The epithelium of the Saccus vasculosus of Perca fluviatilis consists of coronet cells and bipolar liquor contact neurones in a 41 ratio, and supporting cells. The organ also contains macrophages.The coronet cells have been studied after different kinds of fixation. The globules of these cells contain tubelike cisternae, which do not connect with cisternae in the cell's apical protrusion. The cytoplasm of the apical protrusion and to a greater extent of the globules, is stained by longlasting OsO₄-impregnation. The coronet cells have no direct contact with the basement membrane of the organ. They are neither innervated nor have axons.The dendrites of the bipolar nerve cells end with a bulbous structure protruding into the cerebrospinal fluid. The dendrites contain vesicles and each bears 1–2 cilia. The axons join the fiber bundles of the epithelium. There are synaptic contacts on the surface of the neurons and their processes. In some vesicular structures within the apices and more conspicuously within the extracellular space around these cells indications of acetylcholinesterase activity are found.It appears that the nervus sacci vasculosi contains only afferent axons of the bipolar liquor-contact neurons and efferent fibres which form synaptic contacts with these neurons.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

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.     

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.     

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.     

Distribution of the Parathyroid Hormone-Related Peptide and Its Receptor in the Saccus Vasculosus and Choroid Plexus in the Red Stingray (Dasyatis akajei: Elasmobranch)

1. The exact role of the parathyroid hormone-related peptide (PTHrP) is not fully understood. We used immunohistochemistry to localize the PTHrP and its receptor in the brain of the red stingray, particularly in the saccus vasculosus (SV) and choroid plexus.2. Immunoreactive PTHrP and its receptor were detected in the epithelial cells of the SV and the choroid plexus. In addition, the neuronal perikarya in the nucleus of the SV located in the hypothalamus is positive for the PTHrP.3. No PTHrP-containing neurons were detected in the choroid plexus. Extracts of SV and choroid plexus showed positive reactions against the PTHrP and its receptor antibody in Western blot analysis.4. High levels of immunoreactive PTHrP were detected in the plasma equivalent to those present in human humoral malignant hypercalcemia. In contrast, the immunoreactive PTHrP concentration in the cerebrospinal fluid was below detectable levels.5. Our results suggest that the regulation of the PTHrP in the SV differs from that in the choroid plexus in the red stingray.     

Fine structural localization of adenosine triphosphatase activities in the saccus vasculosus of the rainbow trout,Salmo gairdneri Richardson

The following characteristics of the adenosine triphosphatases (ATPase) in the saccus vasculosus were studied in Salmo gairdneri Richardson: 1) distributional pattern, 2) cytochemical properties in relation to different substrates, inhibitors, pH and bivalent metal ions, and 3) ultrastructural localization. Ultracytochemical studies using modifications of the Washstein-Meisel technique showed that within the pH range 7.1-8.0 several Mg++ or Ca++-activated ATPase are localized on the intracellular surface of membranes and in the cytoplasm of ependymal coronet cells and tanycytes ("supporting cells", "Zwischenzellen", glial cells"). The high ATPase activity at the level of the specialized luminal plasma membranes of coronet cell globules and of tanycyte microvilli is discussed in relation to phenomena of active transport and a possible resulting transfer of low-molecular weight substances into and/or from the cerebrospinal fluid (CSF). The localization of ATPase on the specialized membranes of primary vesicles is considered in connection with available structural and enzyme-cytochemical data on a possible function of these cell organelles in storage and release of substances (including Ca++ ions?). The cytoplasmic ATPase activity in coronet cells is ascribed to microtubules and/or possible existing contractile proteins/filaments, presumably concerned with internal transport or motility processes. In tanycytes ATPase activity is believed to be associated with the characteristic microfilamentous system of still unknown function. The ATPase activity in the (9 + 0) ciliary apparatus of globules could not be interpreted in terms of motility. The present study provides further support to the proposed hypothesis of the transport function of the saccus vasculosus, and an extension of the concept in the sense that not only the principal coronet cells, but also the tanycytes of this circumventricular organ are involved in CSF-homeostasis.     

Ultrastructural changes in the coronet cells of the saccus vasculosus from rainbow trout,Salmo gairdneri (Richardson), kept in sea water

Summary The coronet cells of saccus vasculosus of fresh-water living and sea-water adapted rainbow trout were studied with the electron microscope, with special regard to changes in the latter group. Only quantitative differences were observed, namely a raised number of mitochondria in the apical region and the head and also a concentration of the agranular endoplasmic reticulum with a higher amount of electron-dense material and vesicles around the Golgi saccules. Together, these findings suggest a secretory function for the coronet cell. A supposed transport of vesicles from the head region of the coronet cell out into the globules is suggested. Interrelation between primary and secondary vesicles is discussed.This work was supported by grants from the Royal Physiographical Society of Lund and the Faculty of Natural Sciences at the University of Lund.—I am greatly indebted to Mrs. Lena Svenre and Miss Inger Norling for excellent technical assistance.     

Ultrastructural development of the saccus vasculosus in rainbow trout (Salmo gairdneri)

Summary The morphology of crown cells and supporting cells of the saccus vasculosus has been described by numerous investigators. A third type of cell has been mentioned by several authors and referred to variously as undifferentiated crown cells, pseudo-coronet cells, pear-shaped cells and, most recently, as liquor-contact neurons. A developmental study of the organ was undertaken as a possible means of characterizing this third cell type and determining its origin.The epithelium of the saccus vasculosus and the ependyma of the third ventricle are different and distinguishable at the time of hatching in rainbow trout. Initially, apical protrusions from crown cells extend slightly into the lumen and a few end knobs or motile cilia project from them. Basal bodies with cross-striated rootlets occur frequently. In swim-up fry, end knobs are more numerous and heavily vacuolated, although cross-striated rootlets are less apparent.Evidence is presented that is consistent with a hypothesis of secretory activity in the crown cells. Further, portions of end knobs containing this material appear to be pinched off from the remainder of the crown cell. The possible presence of bipolar neurons is also discussed.Supported by Research Grant 5 R01 NS0627 from the National Institute of Neurological Diseases and Stroke.     

Elektronenmikroskopische Beobachtungen am Saccus vasculosus einiger Knorpelfische

Zusammenfassung Das Epithel des Saccus vasculosus zweier Arten von Selachiern und von einem Holocephalen wurde, nach Perfusionsfixierung der Tiere mit Glutaraldehyd, elektronenmikroskopisch untersucht.Krönchenzellen werden von kleineren Stützzellen unterschieden. Beide Zellarten sitzen der Basalmembran auf und begrenzen apikal das Lumen des III. Ventrikels. Das gut entwickelte agranuläre endoplasmatische Reticulum der Krönchenzellen deutet auf eine hohe biochemische Aktivität hin. Feinstruktur, Gestalt und Lage der Krönchenzellen im Epithel sprechen für ihre Rezeptorennatur. Neben Krönchenzellen ragen einzelne birnenförmige Zellen in das Lumen des Saccus vasculosus der Haie. Zahlreiche Vesikel im apikalen Cytoplasma weisen ebenso wie ein großer Golgi-Apparat auf sekretorische Funktion dieser Zellart. Basisnah splittern Bündel markloser Axone in Einzelfasern auf. Synaptische Verbindungen zwischen Krönchenzellen und solchen Axonen wurden nicht beobachtet.

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Electronmicroscopical observations on the Saccus vasculosus of some Chondrichthyes

Summary The Saccus vasculosus of two species of Selachians and one species of Holocephala was investigated with the electron-microscope after perfusion of the animals with glutaraldehyde.Coronet cells are distinguished from smaller polymorphic supporting-cells. Both types of cells extend from the basement membrane to the lumen of the IIIrd. ventricle. Smooth endoplasmic reticulum is well developed in the coronet cells, and indicates a high biochemical activity. Ultrastructure, shape and position of the coronet cells in the epithelium speak in favour of the receptor hypothesis of these elements. Besides coronet cells occur pear shaped cells which project into the lumen of the Saccus vasculosus of the two species of sharks. Their submicroscopical structure points out secretory function of these cells. Bundles of nonmyelinated nerve fibers split into individual fibers near the basement lamina. Synaptic connections between these axons and the coronet cells have not been observed.

Herrn Prof. Dr. H. Leonhardt danke ich für Themenstellung und zahlreiche Anregungen, Herrn Prof. Dr. Dr. h. c. W. Bargmann für einen Arbeitsplatz im Anatomischen Institut Kiel.     

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