Intracellular localisation of phytochrome in oat coleoptiles by electron microscopy

We have analysed the intracellular localisation of phytochrome in oat coleoptile cells by electron microscopy and confirm and extend light-microscopical findings of previous authors. We used indirect immuno-labeling with polyclonal antibodies against 60-KDa phytochrome from etiolated oat seedlings, and a gold-coupled second antibody, on ultrathin sections of LR-white-embedded material. In dark-grown seedlings, phytochrome-labeling is distributed diffusely throughout the cytoplasm. Organelles and membranes are not labeled. After photoconversion of the red-absorbing form of phytochrome to the far-red absorbing form (Pfr) (5-min red light; 660 nm), the label is sequestered uniquely in electron-dense areas within the cytoplasm. These areas are irregularly shaped, are often located in the vicinity of the vacuole, are not surrounded by a membrane, exclude cellular organelles and ribosomes and are not found in dark-grown material; an immediate 5-min farred light pulse after the red light does not cause these structures to disappear. After a dark period of 3–4 h following red-light irradiation, these electron-dense structures disappear together with any specific labeling. We suggest a Pfr-induced aggregation of an unknown, phytochrome-binding protein or proteins.Abbreviations Pr and Pfr phytochrome in its red and far-red absorbing form, respectively     

Cytological observations on the ventromedial hypothalamic nucleus

Two sorts of neurons are recognized in Golgi impregnations of the rat ventromedial hypothalamic nucleus (HVM). The two cell types, category I and II neurons, are differentiated on the basis of their somatic, dendritic, and axonal characteristics. Category I neurons form most of the neuronal population and are located throughout HVM. The small number of category II neurons that have been studied occur in lateral HVM. Two varieties of neuronal profile, "common" and "uncommon cells", are seen in thin sections of HVM. The "uncommon cells", in comparison with the "common ones", appear to have a larger soma, a more electron-dense cytoplasmic matrix, an abundance of Nissl bodies, and a population of dense-cored vesicles (100--130 nm in diameter). Some of the somata and proximal dendrites of "common", but not "uncommon" cells, are wrapped in multiple layers of astrocytic processes. Although the correlation is tentative, it is argued that category I neurons correspond to "common cells" and category II, to "uncommon cells". One possible implication of this correspondence is discussed regarding neuronal alteration in response to change in the endocrinological environment of the brain.     

Über intraepitheliale nervöse Endformationen in mehrschichtigem Plattenepithel

Zusammenfassung Im Anschluß an frühere lichtmikroskopisch erhobene Befunde (Silberimprägnation; 1924, 1928) weist der Verfasser durch elektronenmikroskopische Untersuchungen nach, daß durch die Interzellularspalten des Epithels (Schnauze der weißen Ratte) Nervenfasern hindurchziehen. Diese Nervenfäserchen, ihre Varikositäten und ihre Endknöpfchen liegen in den Spalten zwischen den Epithelzellen. Die Ultrastruktur der nackten, d.h. von Schwannschen Zellen nicht umhüllten terminalen Nervenfasern und ihrer Endigungen im Epithel wird beschrieben.

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On intraepithelial nerve endings in a stratified squamous epithelium

Summary In addition to former light microscopical observations the author demonstrates electronmicroscopically that nerve fibers pass through the intercellular spaces of the epithelium. The nerve fibers, their varicosities and terminal bulbs lie within the spaces between the epithelial cells. The ultrastructure of the terminal nerve fibers in the epithelium is described.

     

Die Feinstruktur des ektopischen Schmelzorgans bei beschleunigter Eruption des Rattenschneidezahnes

Zusammenfassung Durch regelmäßig wiederholtes mechanisches Kürzen des rechten unteren Rattenschneidezahnes wurde eine beschleunigte Eruption hervorgerufen. Nach 15–17 Tagen trat eine Störung der Schmelzentwicklung in Form einer Produktion von kreideweißem statt normal pigmentiertem Schmelz und ein rotbräunliches Häutchen im Bereich des gingivalen Epithelverschlusses in Erscheinung. Dieses Häutchen wurde nach intravitaler Fixierung mit Glutaraldehyd abpräpariert und licht- und elektronenmikroskopisch untersucht.Das Häutchen besteht aus mehreren übereinander gelagerten Zellschichten: aus Ameloblasten, einem typischen Papillarorgan, einer Bindegewebsschicht und aus flachen Deckzellen. Die Ameloblasten im Bereich des Epithelverschlusses befinden sich im Stadium der frühen, die Ameloblasten des aus dem Epithelverschluß hervorwachsenden Häutchens dagegen im Stadium der späten Maturation. Die Ultrastruktur des gesamten Papillarorgans läßt auf eine intensive metabolische Funktion und auf Ionentransport schließen. Pigmentgranula, die bei normaler Eruption im Maturationsstadium sowohl im Papillarorgan als auch in den Ameloblasten vorkommen, wurden nur im Bereiche des Epithelverschlusses in vaskulären Elementen und in perivaskulären Bindegewebszellen gefunden. Die Oberfläche des Häutchens wird von einer schmalen Schicht flacher, organellenarmer Zellen gebildet.Das bei beschleunigter Eruption aus dem Epithelansatz hervorwachsende Häutchen ist ein ektopisches Schmelzorgan, dessen rotbräunliche Färbung von der reichen Gefäßversorgung herrührt.

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The fine structure of the ectopic enamel organ in accelerated eruption of the rat incisor

Summary An accelerated eruption of the right mandibular rat incisor was induced by its repeated, mechanical shortening. Fifteen to seventeen days after beginning of the experiment a disturbance of enamel development occured, manifested by production of white, opaque enamel instead of normally pigmented enamel and by appearance of a redbrownish membrane overlying the enamel. This membrane was intravitally fixed with glutaraldehyde and after dissection processed for histologic and electronmicroscopic investigation.The membrane consists of ameloblasts, a typical papillary organ, a connective tissue layer and of flat surface cells. The ameloblasts near the epithelial attachment are in early maturation state, the ameloblasts of the outgrowing membrane show signs of reduction, typical for late maturation. The papillary organ of the entire membrane showes ultrastructural equivalents of high metabolic activity as well as of fluid transports.Pigment granules, which in normal eruption occur in the maturation state in the papillary cells and in ameloblasts, are absent in these cells of the membrane. They were only seen in perivascular elements and in connective tissue cells near the epithelial attachment. On the oral surface the membrane is covered by a thin layer of flat, degenerated cells poor in organelles. These findings support the concept, that the membrane growing out of the epithelial attachment in accelerated eruption of the rat incisor, is an ectopic enamel organ. The red-brownish colour is not due to pigment accumulation but to its rich vascularization.

     

Die Ultrastruktur der Kapillaren der Kleinhirnrinde und das perikapilläre Gewebe

Zusammenfassung Die Kapillaren im Kleinhirn der Katze haben einen Durchmesser von 3,5–12 . Im Stratum granulosum finden sich vorwiegend engere, im Stratum moleculare und in der Purkinjezellschicht meist weitere Kapillaren. Die Endothelzellen bilden schmale Lamellen, die sich teilweise überlappen und durch tight junctions miteinander verbunden sind. Vom umgebenden Kleinhirngewebe sind sie durch eine Basalmembran abgegrenzt, die sich häufig in zwei Schichten spaltet, zwischen denen Perizyten mit ihren Fortsätzen liegen. Diese sind vornehmlich im Stratum granulosum am Aufbau der Kapillarwand beteiligt.Um die Kapillaren bilden Astrozyten mit ihren Fortsätzen, zum großen Teil aber auch mit ihren Perikaryen, einen unvollständigen Mantel. An den von Astrozyten freien Anteilen der Kapillaroberfläche grenzen Oligodendrozyten, Körnerzellen und Golgizellen mit ihren Perikaryen direkt an die Basalmembran der Kapillaren. Purkinjezellen liegen dagegen nicht unmittelbar der Kapillare an, sondern sind immer durch eine Schicht von Korbzellaxonen und Gliafortsätzen von der Basalmembran getrennt.Kapillaren mit einem Durchmesser von mehr als 10 besitzen einen perikapillären Raum. Dieser ist sowohl gegen die Glia als auch gegen das Endothel der Kapillare durch eine Basalmembran abgegrenzt. Im perivaskulären Raum findet man Perizyten, Fibroblasten und zirkulär verlaufende kollagene Fasern.

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Ultrastructure of capillaries in the cerebellar cortex and the pericapillary compartment

Summary The capillaries in the cerebellar cortex of the cat have a diameter varying from 3.5 to 12 . In the granular layer the capillaries have a smaller diameter than those in the molecular and the Purkinje-cell layer. The endothelium forms slender lamellae which partially overlap. These lamellae are connected with each other by tight junctions. The capillaries are separated from the pericapillary compartment by a basement membrane which often splits into two layers; in between these layers processes of pericytes are located. The pericytes make up a part of the capillary wall mainly in the granular layer.Around the capillaries the astrocytes form an incomplete glial sheath with their processes and also with their pericaryon. Those parts of the capillary basement membrane which are not covered by astrocytes or their processes, are in contact with oligodendrocytes, granule cells or Golgi cells. The Purkinje cells have no intimate contact to the capillary, they are always separated from the basement membrane by a thin layer of basket cell axons and processes of astrocytes.The capillaries with a diameter greater than 10 often have a perivascular space. This space is separated from the endothelium as well as from the nervous tissue by a basement membrane. In the pericapillary space pericytes, fibroblasts and circularly arranged collagenous fibers are located.

Mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft.     

Cellules riches en glycogène dans l'épendyme du ventricule latéral du cerveau de rat

Summary The ependyma of the lateral ventricle of the rat brain was investigated at different ages from 20 days to adulthood.A particular cell type occurs in the external wall of the ventricle, where the proliferative subependymal layer is present. These cells found at all ages studied, are characterized by a high content of glycogen, and a structure different of typical ependymal cells.A large number of nerve endings is situated in close vicinity of these cells, either free in the ventricle lumen, or sometimes ensheathed in the cells. No synapse was found between these endings an the glycogen-rich cells.These glycogen-rich cells undergo several modifications with age: their glycogen content is reduced in the adult, and they acquire a few cilia and gliofilaments. It is suggested that they represent a transitory differentiation of the ependyma, functionally linked with the proliferative subependymal layer.

     

Sensilles énigmatiques de l'antenne de certains Coléoptères

Résumé Nos recherches au microscope électronique portent sur une structure nerveuse en lamelles découverte pour la première fois dans l'antenne duSpeophyes, qu'on étudie comparativement chez les Coléoptères hypogés et épigés. Nous constatons qu'elle existe à l'extrémité de l'antenne chez toutes les espèces cavernicoles de l'échantillon examiné ainsi que chez plusieurs espèces épigées. Nous n'avons pas rencontré cet organe à l'extrémité de l'antenne de trois espèces épigées; nous ne pouvons cependant pas affirmer qu'il n'existe pas, les articles inférieurs de l'antenne n'ayant pas été examinés. Son plus grand développement chez les espèces cavernicoles laisse supposer une fonction plus spécifiquement adaptée à ce milieu. La structure fine de ces lamelles présentant une étroite similitude avec celle des photorécepteurs, nous avons émis l'hypothèse d'une sensibilité à des rayonnements présents dans la grotte, par exemple au rayonnement infrarouge. Il faut souligner que la nature ciliaire de ces structures est en désaccord avec l'appartenance de l'Insecte à la lignée rhabdomérique.

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Comparative study on the lamellated nervous structures in the antenna of certain coleoptera

Summary The lamellated nervous system discovered in the antennae of the beetleSpeophyes, has been comparatively investigated in various hypogeous and epigeous species. It was found in almost all of the species studied in the tip of the antennae. It was, however, not detected in a few of the epigeous forms. Its extensive development in the cave develling species suggests a specific function in this particular environment. The fine structure of the lamellae is closely comparable to that of photosensory cells. Therefore it is assumed that they serve a similar function, i. e. the perception of infrared radiation in the caves. Its ciliary nature does not correspond to the rhabdomeric structure of insect eyes.

Technicienne associée.     

Monoamines in rat thyroid parafollicular cells and the effect of vitamin D2-induced degranulation

Summary Thyroid parafollicular cells of normocalcemic and vitamin D₂-treated rats were investigated by electron microscopy and with the histochemical fluorescence technique of Hillarp and Falck.Administration of high doses of vitamin D₂ caused hypercalcemia and an extensive degranulation of the parafollicular cells.The formation and storage of monoamines in granulated and degranulated parafollicular cells was investigated by fluorescence microscopy after injection of monoamine precursors (DOPA, 5-HTP), alone or in combination with Ro 4-4602, nialamide or reserpine.No fluorescence was observed in parafollicular cells of untreated rats. l-DOPA and l-5-HTP (but not the corresponding D-amino acids) were taken up by a process closely linked to the decarboxylation of the amino acids to the corresponding amines (dopamine and 5-hydroxytryptamine). Treatment with vitamin D₂ did not seem to affect the formation of amines in the parafollicular cells or the formation and storage of amines in other cell systems investigated. The amine itself (dopamine) was not taken up by the parafollicular cells.In normocalcemic rats, the amine formed was retained in the cytoplasm of the parafollicular cells by a partially reserpine-resistant mechanism. The storage of amines is concluded to occur in association with the calcitonin-containing granules.In parafollicular cells of vitamin D₂-treated rats, a certain amount of amine was bound in the cytoplasm in the absence of typical granules. As a considerable amount of calcitonin is known to remain in the thyroid of vitamin D₂-treated rats, the present observations may indicate an association between the amine and the polypeptide hormone calcitonin, whether the latter is confined to typical granules or not.The present study was supported by grants B72-12X-3352-02 and B72-14X-2207-06B from the Swedish Medical Research Council and by grants from Magnus Bergwall's Foundation, Gustav and Majen Lundgren's Foundation, Wilhelm and Martina Lundgren's Foundation and from the Faculty of Medicine, University of Göteborg, Sweden. For skilful technical assistance we are indebted to Mrs. Kirsten Collin and Mr. Pär-Anders Larsson.