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  • 1
    Electronic Resource
    Electronic Resource
    Springer
    Anatomy and embryology 175 (1987), S. 331-340 
    ISSN: 1432-0568
    Keywords: Neural plate ; Neuronal progenitor cells ; Neuron transformation ; In vivo ; In vitro
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Tissue from the mouse neural plate and neural tube was studied, by light and electron microscopy, as starting material for tissue culture. In vivo, up to embryonic day 9 (E 9, stage Th 14; Theiler 1972) all neuroepithelial cells of the neural plate were mitotically active. As judged from their light microscopic or ultrastructural appearance, they could hardly be distinguished from one another or from neuroepithelial cells of more mature embryos. The earliest few immature neurons in the mesencephalic anlage were discernible on day 9 1/2 (stage Th 15) in the prospective intermediate layer of the neural tube, concomitantly with the development of processes containing neurotubules and vesicles which were oriented in parallel to the basal lamina. For tissue culture, explants of the mesencephalic anlage of embryonic days 8 (Th 12/13), 9 1/2 (Th 15), and 11 (Th 18) were kept in vitro and their development was compared with each other and with the corresponding developmental stage in vivo in the initial phase of culture (e.g., E 8, day of explanation, kept in vitro for 2 days, E 10 in vivo being the stage for comparison). The study demonstrated that further in vitro development proceeded in an accelerated manner, independent of the developmental stage of the embryo from which the tissue was explanted. In vitro, proliferation of the explanted neuronal progenitor cells stopped in all explants within 24 h of culture as revealed by autoradiographic and electron microscopic techniques. Cytoplasmic transformation was observed corresponding to that found in vivo, but always greatly accelerated. Earliest axons had formed after 24 h in vitro; synapses with clear vesicles and dense core vesicles were observed after at least 3 days in culture in all explants regardless of age at the time of explantation (E 8 or E 11). The present ultrastructural results indicate that prospective neurons within the neuroepithelium of the neural plate and early neural tube were immediately able to develop into neurons without the complete sequence of mitotic events normally occurring under in vivo conditions.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1471-4159
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Abstract: Expression of receptors for the neuropeptide somatostatin was investigated in vitro in rat and human astrocytes, glioma cell lines, and solid human glial tumors that were all immunopositive for the astrocytic marker glial fibrillary acidic protein. After affinity labelling with a peptide-gold conjugate of known biological activity, somatostatin-binding sites could be visualized at the light- and electron-microscopic level on the surface of glial cells. Glioma cells were generally labeled more strongly than were normal astrocytes and preferentially bound the ligand at their processes and not at their somata as were normal cells. Somatostatin transmembrane receptor (SSTR) subtype expression was probed by reverse transcription-polymerase chain reaction: In rat and human cortical astrocytes and in one glioma cell line (U 118), a pattern of three subtypes (SSTR-1, SSTR-2, and SSTR-4) was detected, whereas, in all other glioma cell lines and in six solid glial tumors investigated, the SSTR-2 subtype was relatively stronger, expressed either alone or in combination with SSTR-1; sometimes SSTR-3 or SSTR-4 was demonstrated in clearly reduced amounts. In astrocytes and gliomas, somatostatin reduced the levels of cyclic AMP elicited by the adenylate cyclase activator forskolin indicating that at least one of the receptor subtypes is negatively linked to adenylate cyclase. In contrast to other cell types, somatostatin did not inhibit the basal or the fetal calf serum-stimulated proliferation of astrocytes, glioma cell lines, or glial tumors in culture. Thus, strong SSTR-2 subtype expression characterizes glial tumors, but somatostatin is ineffective in inhibiting their growth.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 263 (1991), S. 253-263 
    ISSN: 1432-0878
    Keywords: Astrocytes ; Diencephalon ; Receptors, membrane ; Somatostatin (SRIF) ; Rat (Han: Wist)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a somatostatin-gold conjugate of known biological activity, high affinity binding sites for this neuropeptide were visualized at cellular resolution on cultured diencephalic astrocytes and on frozen sections of the rat diencephalon. Binding could be completely suppressed in competition experiments with surplus unlabeled somatostatin. On sections, the ligand was displaced from its binding sites by 10 μM guanosine triphosphate indicating a functional significance of the labeled structures. As with the native peptide, a surplus of the analog SMS 201–995 suppressed nearly all staining. The ligand was bound to distinct populations of astrocytes, namely to those in subependymal and perivascular positions, to astrocytes in somatostatin-innervated hypothalamic nuclei in the mid-sagittal plane and to borderline regions of circumventricular organs. A general mismatch between the distribution of somatostatin-immunoreactive terminals and the pattern of binding of the ligand does not exist. This, together with the competition experiments, suggests a functional relationship between the somatostatin-releasing neurons and associated astrocytes.
    Type of Medium: Electronic Resource
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  • 4
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 262 (1990), S. 431-443 
    ISSN: 1432-0878
    Keywords: Astrocytes ; Telencephalon ; Receptors, membrane ; Somatostatin (SRIF) ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary Using a somatostatin-gold conjugate as ligand, high-affinity binding sites for this neuropeptide were demonstrated at three levels: (i) cultured astrocytes from rat cortex, (ii) hippocampal slice cultures, and (iii) frozen tissue sections of rat telencephalon. The conjugate proved as active as the native peptide in competing for the binding sites. Light-microscopic visualization of bound ligand was achieved by silver intensification of the colloidal gold. This method is faster and yields superior resolution compared with autoradiography. Cultured astrocytes from cortex and hippocampus could be labeled by the ligand. At the light- and electron-microscopic level, astrocytes could be double-labeled by the somatostatin-gold conjugate and immunostaining for glial fibrillary acidic protein (GFAP). In hippocampal slice cultures, the conjugate did not penetrate into the neuropil because of a covering glial layer. However, a portion of this completely GFAP-positive covering glia reacted with the somatostatin ligand. In frozen brain sections, apart from delicate punctate structures, two types of labeled glia cells were seen: single stellate astrocytes and perivascular glia cells.
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  • 5
    ISSN: 1432-0878
    Keywords: Key words: Thymus ; Thymulin ; Epithelial cells ; Cell culture ; Rat (Han:WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract. A new method for the long-term culture of pure rat thymic epithelial cells was established. The cultures were characterized by immunocytochemistry, electron microscopy and proliferation assays. Non-epithelial thymic cells were eliminated with a reliable and reproducible pre-plating method, by differential trypsin treatment of the cultures and by addition of horse serum to the culture medium instead of fetal calf serum. The final cultures contained more than 95% pure epithelial cells as evidenced by immunostaining for cytokeratin. Ultrastructural studies indicated that these cells are physiologically active epithelial cells with tonofilaments, desmosomes and filopods. The subsets of the thymic epithelial cells in vitro were investigated by comparing their staining pattern with that obtained in situ using several subtype-selective antibodies. Thymic epithelial cells in vitro showed a preferential expression of subcapsular/perivascular and medullary markers. Only few cultivated cells were of cortical origin. In the first to the fourth subcultures, some cells were immunopositive for the thymus hormone/factor thymulin. The proliferation of thymic epithelial cells was stimulated by horse serum and to a lesser extend by fetal calf serum. The adenylate cyclase activators isoproterenol and forskolin, and the glucocorticoid cortisol inhibited the proliferation.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1432-0878
    Keywords: Somatostatin ; Cholecystokinin ; Binding sites ; Receptors ; Neurons ; Telencephalon ; Rat (Han: WIST)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract The identification of high-affinity binding sites for neuropeptides on individual target cells is a prerequisite when studying the sites of action and the manner in which peptides act as neuromediators. In situ and in vitro, this can be achieved using newly synthesized, biologically active conjugates of somatostatin or cholecystokinin (sulphated octapeptide) with colloidal gold. Labelled neurons show a peptide-specific, non-overlapping distribution in rat telencephalic structures; i.e, whereas the somatostatin-gold conjugate labels binding sites on neurons and glial cells, cholecystokinin-binding sites are restricted to neurons. Binding of either gold-labelled ligand can be competitively suppressed by excess amounts of the native peptide or its analogues. Neuronal somatostatin-binding sites are visualized on neurons in lamina III and, in particular, in lamina V/VI of the primary somatosensory cortex and in the magnocellular nucleus of the telencephalic cholinergic system. Cholecystokinin-binding sites are localized in the main olfactory bulb, on neurons in the cortical “hindlimb” and “forelimb” region, in the hippocampus, and in the cingulate and visual cortex.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 123 (1972), S. 47-54 
    ISSN: 1432-0878
    Keywords: Neurosecretion ; Neurohypophysis ; Exocytosis ; Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung 1. Elektronenmikroskopische Untersuchungen an der Neurohypophyse von Ratte und Forelle ergeben, daß sich eine Exocytose von Elementargranula an den Endigungen der neurosekretorischen Fasern nur selten abspielt. Es wird daher angenommen, daß die Abgabe von Hormonen in der Neurohypophyse in der Regel nach dem Muster des „membrane-release“ abläuft. 2. Die Exocytose wird nicht durch eine unmittelbare tangentiale Fusion der Membran des Elementargranulums mit dem Plasmalemm der Nervenendigung (Axolemm) eingeleitet. Vor allem bei Anwendung eines Goniometertisches wird erkennbar, daß vor der Exocytose zwischen Axolemm und Membran des Granulums eine Verbindung in Gestalt eines Stieles entsteht. Die Länge dieses Verbindungsstückes entspricht etwa 2 Axolemmdicken. An der Basis des Stiels im Axolemm tritt das Stoma auf, durch das der Inhalt des Granulums bzw. dieses selbst das Axonende verläßt. 3. Die Herkunft kleiner membrannaher Vesikel (Durchmesser 500 Å) in den Endigungen neurosekretorischer Nervenfasern in der Neurohypophyse konnte nicht geklärt werden. Anzeichen einer kompensatorischen Endocytose im Sinne von Nagasawa, Douglas und Schulz (1970) wurden nicht beobachtet.
    Notes: Summary 1. Electron microscopical investigations of the neurohypophysis in rat and trout reveal that exocytosis of neurosecretory elementary granules from the nerve endings occurs only rarely. The authors are of the opinion that hormone release in the neural lobe follows mainly the “membrane-release” pattern. 2. Exocytosis is not performed by tangential fusion of the elementary granule membrane and the plasmalemma of the nerve ending (axolemma). Administering the goniometer technique one can observe the appearance of a stalk-like structure connecting the two membranes. The basis of the stalk in the axolemma corresponds to the site of the stoma through which the core of the vesicle leaves the nerve ending. 3. The mechanism of the origin of small clear vesicles (diameter 500 Å approx.) near the axolemma of the neurosecretory terminal has not been elucidated. The authors did not observe equivalents of a compensatory endocytosis in the vicinity of granules released by exocytosis.
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  • 8
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 142 (1973), S. 241-262 
    ISSN: 1432-0878
    Keywords: Apical organ (Statocyst) ; Pleurobrachia pileus (Ctenophore) ; Cell types ; Cytochemistry, Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Description / Table of Contents: Zusammenfassung Die Statocyste der Ctenophore Pleurobrachia pileus wurde lichtmikroskopisch, histochemisch und elektronenmikroskopisch untersucht. Folgende Ergebnisse, die zum Teil die Feststellungen anderer Autoren ergänzen, wurden erzielt: Die Statocyste besteht aus einem bilateral symmetrischen Zellpolster, das von einer sog. Cupula überwölbt wird. Die Cupula baut sich aus mit „balancer“-Cilien durchflochtenen Zellen auf und enthält schwefelsäureesterhaltige Mukosubstanzen. Unterhalb der Cupula befindet sich die Masse der intrazellulär liegenden Statolithen. In dem unspezifischen Statocystenepithel lassen sich ebenfalls schwefelsäureesterhaltige Mukosubstanzen nachweisen. In ihm kommen ferner mutmaßlich vibrationssensible Cilien mit stark modifizierter, kolbenförmiger Wurzel vor. Die Reaktionsprodukte der alkalischen Phosphatase sind im unspezifischen und spezifischen Statocystenepithel gleichmäßig granulär verteilt. Die vier Zellgruppen am Rand des spezifischen Statocystenepithels, die die zweiwurzeligen „balancer“-Cilien tragen, geben positive Reaktionen auf ATPase, saure Phosphatase und Acetylcholinesterase. Die übrige oberflächliche Zellschicht des spezifischen Statocystenepithels enthält saure Substanzen und trägt feine Cilien mit gespaltener Wurzel. In mittlerer Höhe des spezifischen Statocystenepithels liegen die schon lichtmikroskopisch sichtbaren vier Gruppen der sog. Lamellenkörper, die sich wahrscheinlich aus mehrfach reduplizierten Centriolen entwickeln und keinen Zusammenhang mit der Zellmembran haben. Die Lamellenkörper werden von Zellfortsätzen umgeben, die zahlreiche Sekretgranula und Mikrotobuli enthalten. Die Basis des spezifischen Statocystenepithels ist reich an B-Esterasen und ist durch zwei Zelltypen charakterisiert: dichte, kubische Zellen, die vermutlich Stammzellen darstellen, und langgestreckte, helle Zellen, deren Fortsätze besonders viele Mikrotubuli enthalten. Die Frage nach dem funktionellen Zusammenspiel morphologisch verschiedener Rezeptoren und das Vorkommen eindeutig nervöser Strukturen werden diskutiert. Ein Schema faßt die Befunde zusammen und ergänzt die bekannten schematischen Darstellungen der Statocyste (Hyman, 1940; Barnes, 1963) unter Berücksichtigung der Beobachtungen von Horridge (1963–1969).
    Notes: Summary The statocyst of the ctenophore, Pleurobrachia pileus, was studied by light and electron microscopy and by histochemical techniques. The following results, which partly extend the findings of other authors were obtained: the statocyst consists of a bilateral symmetric cushion-like epithelium, which is arched over by a so-called cupula. The cupula is built up by cells which are interwoven by “balancer”-cilia and contains sulphurylated mucoid substances. Underneath the cupula there is the complex of the intracellular statoliths. In the unspecific epithelium of the statocyst there are sulphurylated mucoid substances as well. In addition it contains presumably vibration sensitive cilia with a strongly modified onionshaped root. In the specific and unspecific epithelium of the statocyst the reaction products of the alkaline phosphatase are distributed evenly and in granular form. In the four groups of cells at the borders of the specific epithelium, bearing the double-rooted balancer-cilia, the reaction products of ATPase, acid phosphatase and acetylcholinesterase were detected histochemically. In the remaining superficial cell layer of the specific epithelium acidic substances occur. In addition it shows fine cilia with split rootlets. As already seen by light microscopy four groups of lamellated bodies are found at a medium level of the specific epithelium. They presumably develop from multiple reduplicated centriols and have no connection with the cell membrane. The lamellated bodies are surrounded by cell processes containing numerous secretory granules and microtubules. The basal layer of the specific epithelium is rich in B-esterases and is characterized by two types of cells: dense, cuboidal cells which presumably are stem cells and elongated clear cells, the processes of which are particularly rich in microtubules. It is discussed wether morphologically distinct receptors are cooperating in the sense of a functional entity and wether there exist typical nervous structures or not. A scheme summarizing the reported results takes into account the recent findings from other laboratories.
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  • 9
    ISSN: 1432-0878
    Keywords: Neurosecretory hypothalamic area (rat) ; Vasopressin, Oxytocin ; Pregnancy ; Thirst ; Immunocytochemistry, Electron microscopy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary In untreated, pregnant and thirsting rats the neurosecretory hypothalamic areas were investigated by means of the immunoperoxidase technique in order to demonstrate vasopressin- and oxytocin containing elements at the light- and electron microscopic level. In addition, chromalum-hematoxylinphloxin (CHP) staining and conventional double staining of ultrathin sections were used. The areas investigated included the anterior and posterior supraoptic nuclei, the paraventricular nuclei, the numerous accessory cell clusters in the region between the tractus opticus and the third ventricle as well as the median eminence. In all nuclei and in the accessory cell clusters, the number of vasopressin-reactive neurons exceeds that of oxytocin-reactive neurons. Compared with the anterior supraoptic nucleus, the posterior supraoptic nucleus and the accessory cell clusters react more heavily to prolonged thirst. In the median eminence the neurosecretory axons display close contacts with the portal vessels not only in its lateral portion but in thirsting animals also around the mid-line. There the internal layer is broadened and vasopressin-positive tanycytic processes reach the external zone. Parasagittally, fine vasopressin-positive material can be traced from the internal layer to small deposits at the portal vessels. In long term thirsting animals the typical feature of swollen axons exhibits a characteristic distribution in the median eminence and renders a distinct positive reaction to anti-vasopressin. The release of peptide hormones from the perikarya and from the axons within the nuclei as well as the mode of release within the median eminence are discussed. The significance of the positive immunostaining of the ependymal tanycytes and of some perikarya of the suprachiasmatic nucleus must be reconsidered by further studies. Supported by the Deutsche Forschungsgemeinschaft (Grant Nr. Kr 569/1) and Stiftung Volkswagenwerk
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Cell & tissue research 217 (1981), S. 531-552 
    ISSN: 1432-0878
    Keywords: Somatostatin fiber projections ; Brain stem ; Spinal cord ; Rat
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary By use of the PAP-immunohistochemical staining technique with serial sections, somatostatin-immunoreactive fiber projections into the brain stem and the spinal cord are described. These projections originate in the periventricular somatostatin-immunoreactive perikarya of the hypothalamus and form three main pathways: (1) along the stria medullaris thalami and the fasciculus retroflexus into the interpeduncular nucleus; (2) along the medial forebrain bundle into the mammillary body; and (3) via the periventricular gray and the bundle of Schütz into the midbrain tegmentum. Densely arranged immunoreactive fibers and/or basket-like fiber terminals are observed within the following afferent systems: somatic afferent systems (nucleus spinalis nervi trigemini, substantia gelatinosa dorsalis of the entire spinal cord), and visceral afferent systems (nucleus solitarius, regio intermediolateralis and substantia gelatinosa of the sacral spinal cord). These projections form terminals around the perikarya of the second afferent neuron. Perikarya of the third afferent neuron are influenced by somatostatin-immunoreactive projections into the auditory system (nucleus dorsalis lemnisci lateralis, nucleus corporis trapezoidei). Furthermore, a somatostatin-immunoreactive fiber projection is found in the ventral part of the medial accessory olivary nucleus, in nuclei of the limbic system (nucleus habenularis medialis, nuclei supramamillaris and mamillaris lateralis) and in the formatio reticularis (nucleus Darkschewitsch, nuclei tegmenti lateralis and centralis, nucleus parabrachialis lateralis, as well as individual perikarya of the reticular formation). Targets of these projections are interneurons within interlocking neuronal chains.
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