Sequences of halobacterial tRNAs and the paucity of U in the first position of their anticodons

The sequence of three tRNAs from Halobacterium cutirubrum have been determined. The sequences of tRNAValGAC and tRNAValCAC differ by only one nucleotide which is in the 5' terminal anticodon position. These tRNAs as well as that of tRNAAlaCGC are compared to other known halobacterial tRNAs. An observed paucity (or absence) of U in the first anticodon position is unique to archaebacterial tRNAs and may be indicative of unusual decoding properties of these organisms.     

Effects of batrachotoxin on membrane potential and conductance of squid giant axons

The effects of batrachotoxin (BTX) on the membrane potential and conductances of squid giant axons have been studied by means of intracellular microelectrode recording, internal perfusion, and voltage clamp techniques. BTX (550–1100 nM) caused a marked and irreversible depolarization of the nerve membrane, the membrane potential being eventually reversed in polarity by as much as 15 mv. The depolarization progressed more rapidly with internal application than with external application of BTX to the axon. External application of tetrodotoxin (1000 nM) completely restored the BTX depolarization. Removal or drastic reduction of external sodium caused a hyperpolarization of the BTX-poisoned membrane. However, no change in the resting membrane potential occurred when BTX was applied in the absence of sodium ions in both external and internal phases. These observations demonstrate that BTX specifically increases the resting sodium permeability of the squid axon membrane. Despite such an increase in resting sodium permeability, the BTX-poisoned membrane was still capable of undergoing a large sodium permeability increase of normal magnitude upon depolarizing stimulation provided that the membrane potential was brought back to the original or higher level. The possibility that a single sodium channel is operative for both the resting sodium, permeability and the sodium permeability increase upon stimulation is discussed.     

The effect of 5-bromodeoxyuridine (BrdU) on cardiac muscle differentiation

Cultured cardiac muscle cells undergo cell division and form beating progeny. Incorporation of BrdU into the nuclei of daughter cells does not suppress their ability to beat and form cross-striated myofibrils. Fluorescence microscopy of clones derived from single beating cells fed with BrdU-treated medium for over 2 weeks reveal cytoplasmic fibrils stainable with fluorescein-labeled antimyosin. The effect of BrdU on the emergence of cardiac muscle phenotype was also investigated by utilizing cardiac myogenic precursor cells from precardiac mesoderm in early embryos (stage 4–stage 9). These studies show that the cardiac myogenic cells fall into the following categories with respect to their ability to express the differentiated phenotype in the presence of BrdU: (1) precardiac mesodermal cells that are inhibited; (2) precardiac mesodermal cells that are not inhibited; and (3) beating cardiac muscle cells that are not inhibited. The entry of precardiac cells from the first category to the second and to the third appears to be unsynchronized.     

Elektronenmikroskopische Untersuchungen am Subcommissuralorgan des Meerschweinchens

Zusammenfassung Im Subcommissuralorgan des Meerschweinchens wird das mehrreihig hochprismatische Ependym von einem wechselnd breiten, gefäßführenden Hypendym unterlagert, das neben Astrocyten, Oligodendrocyten und Ependymfortsätzen Zellen mit den feinstrukturellen Merkmalen der Ependymzellen enthält.Die subcommissuralen Zellen in Ependym und Hypendym bilden verschiedene Arten von Sekret: Von den Cisternen des endoplasmatischen Reticulum schnüren sich helle Sekretsäckchen ab, die das Cytoplasma durchsetzen. Im apikalen Zellbereich konfluieren sie mitunter zu großen, unregelmäßig begrenzten Sekretarealen. Helle Sekretsäckchen liefern auch den Rohstoff für dichte Sekretgranula, die in manchen Zellen vom Golgi-Apparat gebildet werden; die in verschiedenen Varianten vorkommenden Granula sind apikal angehäuft. Vereinzelt anzutreffende Zellen sind mit Sekretvakuolen so dicht angefüllt, daß das Cytoplasma zu schmalen, dichten Stegen reduziert ist; der Zellkern ist pyknotisch. Die Sekretvakuolen enthalten sehr wenig flockiges Material. Im Hypendym konfluieren die Sekretvakuolen zu großen intracellulären Hohlräumen, in die gelegentlich Mikrovilli und Cilien hineinragen. Schließlich ist eine Art von apokriner Sekretion zu beobachten: Manche Ependymzellen besitzen nahezu homogene Protrusionen, die weit in den Ventrikel reichen; isoliert im Ventrikel werden organellenfreie Cytoplasmabereiche gefunden.Die Kapillaren besitzen ein unterschiedlich breites Endothel. Die Basalmembran ist an vielen Stellen aufgeweitet und umschließt kleine, helle Bezirke. Häufig ist ein echter perivasculärer Raum vorhanden; er ist mit ungeordnet liegenden Filamenten oder Kollagenfibrillen angefüllt und enthält gelegentlich Adventitiazellen. In schmalen perivasculären Spalträumen beobachtet man öfters ein Streifenmuster (Periode ca. 50 m); es handelt sich dabei um ausgedehnte, nicht fibrillär gegliederte Kollagenbereiche.Der entlang dem Recessus pinealis dünn ausgezogene supracommissurale Teil des Organs ist nur von Randbündeln der Commissura posterior unterlagert, die ein dünner Gliafilz von der Hirnoberfläche trennt.

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Electron microscope studies on the subcommissural organ of the guinea pig

Summary In the subcommissural organ of the guinea pig the ependyma is built up of several rows of prismatic cells. The hypendyma of varying width contains capillaries plus astrocytes, oligodendrocytes and ependymal cell processes as well as elements showing the structural characteristics of ependymal cells.The subcommissural cells in the ependyma and in the hypendyma form various types of secretory products: Light secretory sacs originating from the cisternae of the endoplasmic reticulum pile up in the cytoplasm. Sometimes they confluate to irregularly lined areas in the apical zone. In several cells the light secretory sacs deliver material for dense secretory granules which are produced by the Golgi apparatus; dense granules of varying shape are accumulated apically. Some cells are tightly filled with secretory vacuoles. The cytoplasm between the vacuoles is condensed and reduced to narrow rims; the nucleus is pyknotic. The secretory vacuoles contain very little fluffy material. In the hypendyma the secretory vacuoles confluate forming giant vacuoles, occasionally containing microvilli and cilia. Finally a kind of apocrine secretion is observed: Some ependymal cells have protrusions which possess a nearly homogeneous cytoplasm and extend far into the ventricular lumen. Isolated cytoplasmic areas lacking organelles are to be found within the ventricle.The endothelium of the capillaries varies in width. At some places the basement membrane is widened and encloses small areas of lower density. Often a true perivascular space is found, filled with disordered filaments or collagen fibrils; occasionally it contains adventitial cells. Sometimes a substance exhibiting a periodic pattern (period ca. 50 m) occurs in narrow perivascular spaces; this material consists of extended areas of non-fibrillar collagen.The thin supracommissural part of the organ extends along the recessus pinealis. The adjoining commissura posterior is flattened to only a few axon bundles which are separated from the cerebral surface by a thin felt of glial processes.

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Herrn Dozenten Dr. phil. Ernst Kinder zur Vollendung seines 60. Lebensjahres gewidmet.

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Die Arbeit wurde mit dankenswerter Unterstützung durch die Deutsche Forschungsgemeinschaft ausgeführt. — Frau H. Asam danken wir für ausgezeichnete Mitarbeit bei der Präparation; ihr und Frl. C. Degen, Frl. I. Dürr und Frau B. Rottmann für die Ausführung der photographischen Arbeiten. Herrn Dr. med. A. Meinel gebührt unser Dank für wertvolle Diskussionen und Mithilfe bei der Fixierung der Objekte.     

Interchain hydrogen bonds via bound water molecules in the collagen triple helix

If the collagen triple helix is so built as to have one set of NH ? O hydrogen bonds of the type N₃H₃(A) ? O₂(B), then it is possible to have a linkage between N₁H₁(B) and O₁(A) through the intermediary of a water molecule with an oxygen O leading to the formation of the hydrogen bonds N₁(B) ? O and O (A). In the same configuration, another water molecule with an oxygen O can link two earbonyl oxygens of chains A and B forming the hydrogen bonds O O₁(A) and O O₀ (B). The two water oxygens also become receptors at the same time for CH ? O hydrogen bonds. Thus, the neighboring chains in the triple helix are held together by secondary valence bond linkages occurring regularly sit intervals of about 3 Å along the length of the protofibril. The additional water molecules occur on the periphery of the proto-fibril and will contribute their full share towards stabilizing the structure in the solid state. In solution, they will be disturbed by the medium unless they are protected by long side groups. It appears that this type of two-bonded structure, in which one NH ? O bond is to a water molecule, can explain several observations on the stability and hydrogen exchange properties of collagen itself and related synthetic polypeptides. The nature of the water bonds and their strength are found to be better in the one-bonded structure proposed from Madras than in the one having the coordinates of Rich and Crick.