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.

---

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.     

Electromotility of outer hair cells from the cochlea of the echolocating bat,Carollia perspicillata

Isolated outer hair cells (OHCs) and explants ot the organ of Corti were obtained from the cochlea of the echolocating bat, Carollia perspicillata, whose hearing range extends up to about 100 kHz. The OHCs were about 10–30 m long and produced resting potentials between-30 to -69 mV. During stimulation with a sinusoidal extracellular voltage field (voltage gradient of 2 mV/m) cyclic length changes were observed in isolated OHCs. The displacements were most prominent at the level of the cell nucleus and the cuticular plate. In the organ of Corti explants, the extracellular electric field induced a radial movement of the cuticular plate which was observed using video subtraction and photodiode techniques. Maximum displacements of about 0.3–0.8 m were elicited by stimulus frequencies below 100 Hz. The displacement amplitude decreased towards the noise level of about 10–30 nm for stimulus frequencies between 100–500 Hz, both in apical and basal explants. This compares well with data from the guinea pig, where OHC motility induced by extracellular electrical stimulation exhibits a low pass characteristic with a corner frequency below 1 kHz. The data indicate that fast OHC movements presumably are quite small at ultrasonic frequencies and it remains to be solved how they participate in amplifying and sharpening cochlear responses in vivo.Abbreviations BM basilar membrane - FFT fast Fourier Transfer - IHC inner hair cell - OHC outer hair cell     

Preparation of uniform haemoglobin free human erythrocyte ghosts in isotonic solution

A method is described for the preparation of haemoglobin free human erythrocyte ghosts in isotonic solutions using dielectric breakdown technique. In this single haemolytic procedure, almost complete removal of haemoglobin (? 0.1%) was achieved by subjecting the erythrocytes suspended in phosphate buffered, isotonic KCl solution at 0°C to three consecutive electrical field pulses of 16 kV/cm in the presence of 10 mM EDTA; EDTA was used to prevent electrical haemolysis. Haemolysis is induced by subsequent dilution with isotonic and isoionic solution to lower the EDTA concentration. Haemolysis is complete after 5 min; the cells are centrifuged, washed and resuspended in a solution of the same composition and osmolarity containing 4 mM MgCl₂, but no EDTA. The resealing process, carried out at 37°C, was complete in about 1 h. Measurements of the size distribution of the ghost cells in the hydrodynamically focusing Coulter Counter at varying field strengths in the orifice revealed that the ghost population is nearly uniform. The mean (modal) volume of the ghost cells was 110–120 $\text{μ}\text{m}{}^3$ when suspended in phosphate buffered NaCl solution. The apparent breakdown voltage was about 1.3 V.     

Electric field-mediated cell fusion

The use of electrical fields to guide, hold and fuse cells is described. The electrical fusion process consists of two steps: the cells are collected to form pearl-chains between Pt electrodes by the action of dielectrophoresis, then a brief DC pulse is applied, such that the breakdown voltage of the membranes is briefly exceeded and cell-to-cell juncture of the membranes occurs around the pore formed by the pulse. Giant fused cells (diameter up to 100 m) can be formed by the electrically mediated fusion of mouse 3T3 fibroblast cells, provided that pronase is added just before field application.     

Mechanical stabilization of guard cell protoplasts of Vicia faba

Guard cell protoplasts of Vicia faba were immobilized in cross-linked Ca-alginate. No visible morphological changes were detected under the light microscope over a period of 14 days. The entrapped cells reacted normally to changes of the external osmolarity by shrinking and swelling. Addition of the calcium complexing agent, citrate, led to dissolution of the matrix. After reequilibration with Ca ions the released cells regained their ability to swell and shrink in response to external stress. The released protoplasts could be stained with the vital dye, neutral which was accumulated in the vacuoles. It should also be noted that the protoplasts can be transported when immobilized.     

Distribution of a Fatty Acid cyclase enzyme system in plants

Extracts from tissues of 24 plant species were tested for the enzyme that catalyzes the conversion of 13-l-hydroperoxy-cis-9,15-trans-11-octadecatrienoic acid to the cyclic fatty acid 12-oxo-cis-10,15-phytodienoic acid. The enzyme was detected in 15 of the 24 tissues examined, and was demonstrated in seedlings, leaves, and fruits.     

VESICULAR STORAGE AND RELEASE OF ACETYLCHOLINE IN TORPEDO ELECTROPLAQUE SYNAPSES

Abstract— The disposition of newly synthesized ACh subsequent to depletion of vesicular endogenous ACh by stimulation was studied in the electromotor nerve terminals of Torpedo marmorata using [³H]acetate as a precursor of ACh. Little vesicular [³H]ACh could be isolated from tissue immediately after stimulation at 1 Hz. After 3 h post-stimulation recovery the newly synthesized [³H]ACh is found predominantly in a subpopulation of vesicles distinct from the vesicles containing most of the endogenous poorly labelled ACh. Restimulation of the tissue causes release of highly labelled ACh with a specific radioactivity (SRA) comparable to that of the newly synthesized [³H]ACh in the highly labelled subpopulation of vesicles and significantly greater than the SRA of ACh in the main vesicular pool or the total tissue.     

Penetration and entrapment of large particles in erythrocytes by electrical breakdown techniques

Human erythrocytes suspended in isotonic solutions were subjected to haemolysis by application of an electric field pulse to the cell suspension. The field strengths used were 12 and 16kV/cm, respectively; the pulse duration 40 microseconds. The lysed cells showed resealing properties. The permeability change of the membrane generated by the field pulse and by the subsequent osmotic processes were large enough to facilitate the penetration and entrapment of ferritin and Latex particles (diameter: 0.091 and 0.176 micron, respectively) as revealed by electron microscopy. Correct identification of the Latex particles in the electron-micrographs indicated that LOYTER et al. [J. Cell Biol. 66, 292 (1975)], who recently demonstrated the entrapment of Latex spheres in erythrocytes prepared by osmotic haemolysis mistook electron-dense bodies probably consisting of denaturated protein for Latex particles. Under conditions of osmotic haemolysis, carried out according to BODEMANN and PASSOW, particles could only occasionally be detected within the membrane itself and never within the cell interior, suggesting that the electrical haemolysis method is much more effective in the generation of large holes in the membrane.