Ultrastructural studies of severed medial giant and other CNS axons in crayfish

Summary The distal stumps of severed medial giant axons (MGAs) and of non-giant axons (NGAs) in the CNS of the crayfish Procambarus clarkii show long-term (5–9 months) survival associated with disorientation of mitochondria and thickening of the glial sheath. However, the morphological responses of the two axonal types differ in that neither the proximal nor the distal stump of severed MGAs ever fills with mitochondria as is observed in some severed NGAs. Furthermore, the adaxonal glial layer never completely encircles portions of MGA axoplasm as occurs in many severed NGAs; in fact, ultrastructural changes in the adaxonal layer around severed MGAs are often difficult to detect. No multiple axonal profiles are ever seen within the glial sheath of the proximal or distal stumps of severed MGAs whereas these structures are easily located within severed NGAs.This work was supported by NIH research grant #NS-14412 and an RCDA#00070 to GDB     

Centriole replication during ciliogenesis in the chick tracheal epithelium

Zusammenfassung Direkte Belichtung oder Lichtreizung der Lateralaugen lösen im zentralen Teil der Taubenepiphyse keine elektrische Aktivität aus. Im Stielabschnitt sind dagegen kleine, durch Licht nicht beeinlußbare Dauerentladungen zu beobachten. Die Lage der Ableitungspunkte wurde durch den histologischen Nachweis der elektrolytisch gesetzten Eisenmarken bestimmt. Diese Ableitungsstellen befinden sich im Epiphysenparenchym. Ein stärkeres versilberbares Nervenbündel verläuft im Stiel der Epiphyse. Adrenerge Nervenfasern, die grün fluoreszieren, begleiten die Kapselgefäße und dringen mit diesen in das follikuläre Epiphysengewebe ein. Das Epiphysenparenchym zeigt nach Falck-Hillarp eine Gelbfluoreszenz mit umschriebenen apikalen oder basalen Anreicherungen der fluoreszierenden Substanz. Mit Reserpin kann nur ein Teil dieses Materials entspeichert werden. Elektronenmikroskopisch lassen sich in der Taubenepiphyse keine typischen pinealen Rezeptorzellen nachweisen. Regelmäßig lamellierte Außenglieder fehlen vollständig. Es finden sich lediglich cytoplasmareiche bulböse Zilien und zirkuläre Lamellenkomplexe, die in Tubuli übergehen. Die letzteren können sich mit einem dichten körnigen Material füllen. An einigen Stellen haben solche Lamellenverbände einen Zusammenhang mit keulenförmig verdickten Zilien. Diese Zone ist aber anders strukturiert als das für retinale und pineale Lichtsinneszellen charakteristische Verbindungsstück. Die Zeichen der sekretorischen Aktivität der Taubenepiphyse sind überzeugender als die für einen lichtempfindlichen Sinnesapparat sprechenden Strukturmerkmale. Ein bestimmter Typ der Pinealocyten ist reich an 800–1200 Å großen granulierten Vesikeln, die sowohl am apikalen als auch am basalen, auf die Blutgefäße ausgerichteten Zellpol angehäuft sein können. An den manchmal gelappten basalen Endfüßen sind Zeichen der Ausschleusung des Vesikelinhalts bzw. eines Materialschwundes aus dem granulierten Vesikelkern zu erkennen. Die sekretorische Aktivität der Taubenepiphyse wird mit Hinweis auf die in ihr reichlich vorkommenden 5-Hydroxyindole (Quay) und das Problem der Melatonin-Bildung diskutiert. Erörtert wird auch die Möglichkeit einer Stimulierung dieser Sekretion über den Sympathikus.

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Ultrastructural and functional studies of the pineal organ in the pigeon (Columba livia)

Summary Direct illumination or stimulation with light of the lateral eyes induced no electrical activity in the central part of the pineal organ in the pigeon. In the pineal stalk, however, a train of impulses was observed that failed to respond to changes of illumination. The position of the tip of the recording electrode (iron deposit), was histologically clearly determined to be in the pineal parenchyma. A distinct nerve bundle could be traced down the pineal stalk. Adrenergic nerve fibers showing a green fluorescence accompanied pineal capsular vessels into pineal follicular tissue. Using the method of Falck-Hillarp, the pineal parenchyma showed high concentrations of yellow fluorescent material in the apical or basal parts of the pinealocytes. Only a part of the fluorescent material could be dispersed with reserpine. In the electron microscope, no typical photoreceptor cells were identified in the pineal organ. Only large bulbous cilia, and circular lamellar or tubular complexes, were observed; they sometimes contained a dense granular material. Occasionally, the lamellar complexes had a direct relation to cilia of club-shaped appearance. Signs of secretory activity were more definitive in the pigeon pineal organ than sensory structures. One type of pinealocytes contained many 800–1,200 Å diameter granular vesicles concentrated in the apical or basal (apposed to blood vessels) cell parts. At the occasionally lobulated basal end-feet, a disappearance of the content of the granular vesicles indicated a release mechanism. This secretory activity was discussed in relation to the 5-hydroxyindole content of the pigeon pineal organ (Quay) and the general problem of melatonin synthesis; also, the possibility was considered that the secretory activity may be stimulated by sympathetic nerve fibers.

Ob die von Mihail und Molnár (1965) bei der Taube beschriebenen Epiphysektomie-effekte (Wirkung auf den Inselapparat und Zuckerstoffwechsel) auch in diesen Funktionskreis gehören, muß erneut geprüft werden.     

Fine structural analysis of palpal receptors in the tick Amblyomma americanum (L.)

Summary Palps of the tick Amblyomma americanum (L.) (Acarina: Ixodidae; nymphal stage) were studied by scanning and transmission electron microscopy. The terminal palp segment (IV) bears the so-called palpal organ, a cluster of 10 short, blunt-tipped sensilla. All sensilla (except for the center sensillum) receive a dual innervation: 2 mechanoreceptive dendrites which terminate in the socket membrane plus several chemoreceptive dendrites (4–12) which enter the lumen. The thick-walled cuticular shaft possesses 2–3 small pore openings (100 Å) below the tip, thus establishing communication between dendrites and environment. Two structurally different types of palpal sensilla exist: The A-type has a characteristic doublelumen and always contains 4 dendrites, the B-type features a single lumen and a specially layered cuticular shaft with 6–12 dendrites. The fine structure of the tick palpal receptors corresponds closely to that of known contact chemoreceptors in insects.This research was supported in part by a contract with the Office of Naval Research (R. C. Axtell, principal investigator), and by NIH Training grant ES 00069. Paper no. 3700 of the Journal Series of the North Carolina State University Agricultural Experiment Station, Raleigh.     

Ultrastructure de l'épithélium tégumentaire de Glossiphonia complanata (L.) (Hirudinée): cellules épithéliales et organes sensoriels de Bayer

Résumé Le tégument de Glossiphonia complanata comprend, outre les cellules épithéliales banales, de nombreux organes particuliers ou organes de Bayer, regroupés essentiellement sur la face dorsale de l'animal. Ils sont formés par une cellule apicale saillante enchassée dans une cellule musculaire en anneau.L'ensemble de ces formations est étudié du point de vue ultrastructural. Des cellules épithéliales partent des fibres nerveuses afférentes, sans doute vecteur des perceptions de stimuli mécaniques de pression au niveau du tégument; la réponse se faisant sans doute par la contraction de la cellule basale de chaque organe de Bayer, innervée par des fibres nerveuses efférentes, entrainant la saillie de la cellule apicale. Le hérissement de ces nombreuses papilles du tégument dorsal pourrait être un signal perçu par le partenaire sexuel, chez cette Hirudinée à fécondation hypodermique.

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Ultrastructure of the integumentary epithelium of Glossiphonia complanata (L.) (hirudinea): Epithelial cells and sensory organs of bayer

Summary The integument of Glossiphonia complanata, built up by epithelial cells, contains numerous particular organs (Bayer organs), mainly on the dorsal side of the animal. They consist of a protuberant apical cell, which is surrounded by a ring-shaped muscle cell. All the integumentary formations are studied from an ultrastructural point of view. From the epithelial cells issue afferent nerve fibres, considered as vectors of the perception of mechanical stimuli of pressure at the level of the integument; the response no doubt operating by the contraction of the basal muscle cell of each Bayer organ, innervated by efferent nerve fibres, bringing forth the protrusion of the apical cell. The erection of these numerous papillae of the dorsal integument might be a signal perceived by the sexual partner, fecundation occurring in this group of leeches under the integument.

     

Les cellules sensorielles des papilles de la trompe chez Glycera convoluta Keferstein (Annélide Polychète)

Résumé Chaque papille de la trompe chez Glycera convoluta contient un groupe central de deux à quatre cellules sensorielles primaires. Chaque cellule possède un noyau allongé situé à mi-hauteur de la papille. Le processus péripherique ou dendritique porte cinq à six cils qui traversent la cuticule et sont entourés chacun par un cercle de microvillosités; les cils se prolongent dans le cytoplasme par de fines racines ciliaires qui se réunissent pour former une seule énorme racine montrant une striation périodique. Le processus central ou axonal se prolonge dans le plexus nerveux sous-épithélial. Le rôle physiologique de ces cellules est discuté.

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Sensory cells of the papillae in the proboscis of Glycera convoluta keferstein (Annelida, Polychaeta)

Summary Each papilla in the proboscis of Glycera convoluta contains a central group of two to four primary sensory cells. Each one shows an elongated nucleus situated at half-height of the papilla. The peripheric or dendritic process bears five to six cilia traversing the cuticle; each of them is circled by a ring of microvilli. The cilia are prolonged in the cytoplasm by fine ciliary rootlets which gather themselves in a very big single root showing a periodic striation. The central or axonal process runs in the subepithelial nervous plexus. The physiological role of the cells is discussed.

     

Sensory innervation of the tentacles of the polychaete,Sabella pavonina

Summary Following observation of conical groups of stiff, but motile cilia on the tentacles of the branchial crown of Sabella pavonina, these were examined with the electron microscope. The bundles consist of about 40 unenclosed standard cilia supported by one or two primary sense cells with centrally directed axons of 0.1–0.2 diameter. Axons in the distal portions of the branchial crown occur in small bundles surrounded by a basement membrane. More centrally, glial elements appear and the nerves are surrounded by a collagenous sheath. The branchial nerve trunk shows similarities in organisation to other previously investigated annelid central nervous tissue in that the whole nerve is surrounded by a fibrous sheath central to which there is a layer of glial cells with processes penetrating a central neuropile. The 0.1–0.2 axons commonly occur in glial-enveloped groups of < 40 whilst other axons of larger and mixed diameter are found together.Each tentacle has two branchial nerves on the oral side, and each nerve gives rise to two small 75-axon branches running to each pinnule. The branchial nerves fuse to form the branchial nerve trunk running to the supra-oesophageal ganglia.Sections of the branchial nerves of the branchial crown at progressively more central levels show that the branchial nerve trunk contains enough axons of 0.1–0.2 diameter to account for all the sensory cells on the tentacles. This is taken as evidence for the sensory cells having axons terminating within the central nervous system and that there is no peripheral confluence or fusion of these afferent axons.     

Weitere elektronenmikroskopische Untersuchungen am Pinealorgan vonPhoxinus laevis (Teleostei,Cyprinidae)

Zusammenfassung Auch im Pinealorgan adulter Elritzen (Phoxinus laevis) sind die Außenglieder der Sinneszellen nicht degeneriert. Der Vergleich wurde zwischen adulten Exemplaren (8,6 cm) und 19,5 Monate jüngeren Tieren (3 cm) durchgeführt. Im Verhältnis zu den Jungtieren sind die Neuropilformationen und die synaptischen Strukturen adulter Elritzen stärker ausgeprägt. Bei den letzteren enthalten die Endfüße der pinealen Rezeptoren zahlreiche synaptische Bänder und 300–400 Å große Bläschen. Die Neuropilzonen sind außerdem reich an verdichteten Membranstellen (tight junctions). Im Pinealorgan des adulten Phoxinus finden sich eigenartige Einschlußkörper, die aus konzentrischen Lamellen bestehen; ähnliche Membran wirbel hat Takahashi (1969) bei zweijährigen Goldfischen beschrieben. Häufig lassen sich in den Epiphysen adulter Elritzen Axone beobachten, die mit 500–1000 Å großen granulierten Vesikeln gefüllt sind und im Verband bahnartig zusammengefaßter markloser Nervenfasern verlaufen. Die Herkunft und Bedeutung dieser granulierten Fasern werden diskutiert, ebenso wie die funktionelle Rolle des pinealen Sinnesorgans.

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Further electron microscopic studies of the pineal organ inPhoxinus laevis (Teleostei, Cyprinidae)

Summary The outer segments of pineal receptor cells of adult (8.6 cm) European minnows (Phoxinus laevis), like the lamellated outer segments of 19.5 months younger (3 cm) specimens, did not show signs of degeneration (see also Oksche and Kirschstein, 1967). Neuropile formations and synaptic structures were more prominent in adult than in youngPhoxinus. In the adult group, the end-feet of pineal receptors contained numerous synaptic ribbons and vesicles (300–400 Å in diameter). The neuropile zones were rich in tight junctions. A peculiar structure of the pineal organ of agingPhoxinus was the inclusion body formed by concentric arrangement of lamellae. Membrane whorls similar to these inclusion bodies were described in the pineal organ of two years old goldfish (Takahashi, 1969). In the pineal organ of adultPhoxinus considerable numbers of axons containing dense-core vesicles (500–1,000 Å in diameter) were observed within tract-like bundles of unmyelinated nerve fibers. The origin and significance of these granulated fibers and the functional role of the pineal sense organ have been discussed.

Mit Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Neurofilaments and microtubules in sensory neurons after peripheral nerve section

Summary Sensory neurons were examined in spinal ganglia of the rat 1 to 55 days after section of the plexus brachialis nerves. Only light neurons of the type A were investigated. Maximal reaction to axotomy was found 7 to 14 days after the operation. The majority of the axotomized perikarya developed central chromatolysis. In such neurons, Nissl bodies virtually disappeared from the central area of the neuron and formed a more or less continuous zone at the cell circumference. The cytocentrum became filled with large numbers of mitochondria, dense bodies and other organelles. Neurofilaments and microtubules were disarranged and ran at random among the accumulated particles. Microtubules were often more prominent in chromatolytic areas than neurofilaments. Both these organelles were rare in the peripheral areas filled with massed Nissl substance.Part of the neurons that did not show typical chromatolysis contained increased numbers of neurofilaments among Nissl bodies dispersed throughout the cytoplasm. Neurofilaments were roughly arrayed in bundles up to several microns wide; they were linked by cross-bridges and separated by distances of about 500 Å. Microtubules were rarely found in the filamentous areas. However, they were numerous in the axon hillock and in the initial segment where they formed fascicles similar to those described in normal neurons of other types.During the period from 14 to 55 days after axotomy, many perikarya recovering from chromatolysis contained enlarged bundles of neurofilaments with occasional microtubules among the restored Nissl bodies.Mean diameters of sensory neurons, measured 7 to 55 days after axotomy, in no instance exceeded those of contralateral control neurons. It thus appears that sensory perikarya do not increase in size either during the chromatolytic process or during the period of recovery.This project was supported by a grant from the Muscular Dystrophy Association of America, Inc. The main part of this study was done while the author was a Research Fellow in Anatomy at the Harvard Medical School, Boston. The author wishes to thank prof. S. L. Palay for his valuable advice and help received during her stay at the Department of Anatomy at the Harvard Medical School, under NIH training grant NBO5591.     

Uptake of Endoneurial Lipoprotein into Schwann Cells and Sensory Neurons Is Mediated by Low Density Lipoprotein Receptors and Stimulated After Axonal Injury

During Wallerian degeneration of rat sciatic nerve, the expression of apolipoprotein E increases and apolipoprotein E-containing endoneurial lipoproteins accumulate in the distal nerve segment. In established primary cultures dissociated from dorsal root ganglia, Schwann cells and sensory neurons internalized rhodamine-labeled lipoproteins isolated from crushed rat sciatic nerve as well as low density lipoprotein (LDL) from human serum. The uptake of endoneurial lipoproteins could be inhibited by an excess of LDL or at low temperature (4 degrees C). After transection of nerve fibers in dorsal root ganglia explant cultures, the uptake of lipoproteins was markedly stimulated in Schwann cells that were in close proximity to degenerating neurites. A specific monoclonal antibody directed to the bovine LDL receptor (clone C7) was shown to cross-react with LDL receptor preparations of rat endoneurial cells. LDL receptor immunoreactivity was expressed by cell bodies and processes of cultured Schwann cells, sensory neurons, and fibroblasts from dorsal root ganglia. Incubation of Schwann cells and neurons with the LDL receptor antibody strongly inhibited the uptake of endoneurial lipoproteins. Our results provide direct evidence for the important role of the LDL receptor-mediated pathway to internalize endoneurial lipoproteins into Schwann cells and peripheral neurons required for reuse of cholesterol and other lipids in myelin and plasma membrane biogenesis during nerve repair.