Histochemical and immunohistochemical properties of skeletal muscle fibres fromRana andXenopus

Summary Modern histochemical and immunohistochemical techniques have been used to type skeletal muscle fibres from threeRana species andXenopus laevis.Differing myosin properties and metabolic capacities (representing various contractile properties) define a minimum of four fibre types inRana and five inXenopus. TheRana andXenopus types are sufficiently similar so that a single nomencclature can be applied to them. This nomenclature uses an initial letter indicating the probable contractile performance (F=fast-twitch, S=slow-twitch and T=tonic), and a number indicating rank order of presumed shortening velocity.The largest, fastest fibres-F1-have low oxidative and, at best, moderate glycolytic capacities. Commonly adjacent to them are smaller, F2 fibres with variable but at least moderate metabolic capacities. F3 fibres are rarer and have on average the highest oxidative capacity, and at least moderate glycolytic capacity. They usually occur in the reddest parts of the muscle and, inRana, only in the vicinity of tonic fibres.Metabolically weak, classical amphibian tonic fibres (T5) occur in bothXenopus andRana, but onlyXenopus also has an S4 fibre type. This has moderate metabolic capacity and myosin properties suggesting it is probably capable of slow shortening as well as tonic hold. Immunohistochemically, S4 fibres are most similar to avian slow-twitch fibres.     

Arrangement of connective tissue fibres in the musculus lateralis of the spiny dogfish,Squalus acanthias L. (chondrichthyes)

Summary In the m. lateralis of Selachii a well-developed perimysium is found, while a similar structure is lacking in Teleostei. The Selachian perimysial fibres are arranged in distinct planes. In such planes two predominant fibre directions are found; the angle between each of them and the body-axis is 60–70°. Additionally the myosepts contain fibre bundles which are arranged in oblique loops. These myoseptal fibre-loops are present in Squalus, well as in the Teleosts we examined. In order to investigate the functional significance of these fibre arrangements we constructed a model, consisting of a steel strip representing the axial skeleton and a bundle of stretched rubber tubes representing the lateral musculature. Both components of this model have to be firmly interconnected. This is easily achieved by winding oblique loops of cotton thread around them, thus tying them to each other. When we now release the stretched rubber tubes, they contract, resulting in a proper bending of the model. An optimal value for the angle between the body-axis and the direction of the fibres is found.On the basis of the morphological similarity between the perimysial and the myoseptal loop fibres on the one hand and the cotton thread loops on the other, it is suggested that a functional similarity exists as well. So it is suggested that the functional significance of the perimysial fibre system, and of the myoseptal fibre loops, is the achievement of a firm contact between the lateral muscle mass and the axial skeleton.     

Quantitative analysis of histochemical and immunohistochemical reactions in skeletal muscle fibres ofRana andXenopus

Summary Intensities of histochemical and immunohistochemical reactions in muscle fibres ofRana andXenopus have been estimated microphotometrically, and the data from serial sections statistically analysed. Quantitative validities of reactions and measurements have also been assessed against independent published evidence. It is concluded that NADH—tetrazolium reductase overestimates tonic-fibre aerobic capacities and the actomyosin ATPase reaction overestimates their contraction speeds. However, it appears that succinate dehydrogenase, despite being a near-equilibrium enzyme of particulate distribution, indicates the relative aerobic capacities of fibres with acceptable accuracy when lightly reacted. Capacities for aerobic and anaerobic metabolism are positively correlated over all types of fibre (r typically 0.6 for 200 fibres), perhaps as an adaptation to environmental hypoxia.Multivariate clusters (indicating fibre types) have been sought, using Ward's method with optimizing procedures (iterative relocation and multivariate-normal modelling). Cluster analysis confirms the subjective identifications of two slow/tonic types inXenopus (labelled T5 and S4) but of only one (T5) inRana. Division of the fast family twitch fibres into three types (F1–F3) in both genera, with metabolic capacity related inversely to apparent shortening velocity, is highly supportable by objective criteria. However, statistically significant subdivisions also present themselves.Rana F2 andXenopus F1 clusters can be bisected according to metabolic capacity; andXenopus F2 fibres fall into three subtypes reflecting different isomyosin contents. In the different types of twitch fibre, ratios of myofibrillar ATP consumption rate to aerobic capacity increase up to 30-fold with contraction speed, but anaerobic/aerobic ratios do so only 5-fold.     

Lemdana latifi n. sp. (Filarioidea: Onchocercidae) from the red jungle fowl (Gallus gallus spadiceus)

Lemdana latifi n. sp. was found in connective tissues around the trachea and crop and in the body-cavity of seven of 14 Malayan red jungle fowl Gallus gallus spadiceus. The new species is described and illustrated. Morphologically it is most closely related to Lemdana pavonica and Lemdana francolini. Lemdana latifi is distinguished from the eight valid species of Lemdana by the mean spicular ratio of 1.7:1; the right spicule with a right margin 18–29% (15–31 m; mean 24 m) longer than the left margin; the distal half of the left spicule twisted and S-shaped; and the absence of unpaired papillae at tip of male tail. The new species has smaller adults, a shorter left spicule and a shorter glandular oesophagus than those of L. pavonica and a wider male, shorter spicules and a longer muscular oesophagus than those of L. francolini. The male of L. latifi is 7–9 (8.1)mm long, the left spicule 164–215 (184)m long and the right spicule 98–117 (108)m long. The female is 17–23 (21)mm in length. Sheathed microfilariae from blood smears are 78–100 m long and those from the uterus are 89–103 m long. This is the sixth valid species of Lemdana in the Phasianidae.     

Muscle fibre types and metabolism in post-larval and adult stages of notothenioid fish

Summary A histochemical study was carried out on muscle fibre types in the myotomes of post-larval and adult stages of seven species of notothenioid fish. There was little interspecific variation in the distribution of muscle fibre types in post-larvae. Slow fibres (diameter range 15–60 m) which stained darkly for succinic dehydrogenase activity (SDHase) formed a superficial layer 1–2 fibres thick around the entire lateral surface of the trunk. In all species a narrow band of very small diameter fibres (diameter range 5–62 m), with only weak staining activity, occurred between the skin and slow fibre layer. These have the characteristics of tonic fibres found in other teleosts. The remainder of the myotome was composed of fast muscle fibres (diameter range 9–75 m), which stain weakly for SDHase, -glycerophosphate dehydrogenase, glycogen and lipid. Slow muscle fibres were only a minor component of the trunk muscles of adult stages of the pelagic species Champsocephalus gunnari and Pseudochaenichthys georgianus, consistent with a reliance on pectoral fin swimming during sustained activity. Of the other species examined only Psilodraco breviceps and Notothenia gibberifrons had more than a few percent of slow muscle in the trunk (20%–30% in posterior myotomes), suggesting a greater involvement of sub-carangiform swimming at cruising speeds. The ultrastructure of slow fibres from the pectoral fin adductor and myotomal muscles of a haemoglobinless (P. georgianus) and red-blooded species (P. breviceps), both active swimmers, were compared. Fibres contained loosely packed, and regularly shaped myofibrils numerous mitochondria, glycogen granules and occasional lipid droplets. Mitochondria occupied >50% of fibre volume in the haemoglobinless species P. georgianus, each myofibril was surrounded by one or more mitochondria with densely packed cristae. No significant differences, however, were found in mean diameter between fibres from red-blooded and haemoglobinless species. The activities of key enzymes of energy metabolism were determined in the slow (pectoral) and fast (myotomal) muscles of N. gibberifrons. In contrast to other demersal Antarctic fish examined, much higher glycolytic activities were found in fast muscle fibres, probably reflecting greater endurance during burst swimming.     

A successive histochemical staining for succinate dehydrogenase and “reversed”-ATPase in a single section for the skeletal muscle fibre typing

Summary A procedure is described which simplifies the classification of skeletal muscle fibres in that it allows a simultaneous evaluation of both the oxidative capacity and the intensity of reversed ATPase of the fibres, and thus enables to distinguish three fibre types — SO, FOG and FG — in one tissue section. After preincubation at pH 4.1–4.2 the cryostat section is incubated for succinate dehydrogenase (SDH) and subsequently for reversed-ATPase. This is followed by the fixation with neutral buffered formaldehyde. The results of typing of chicken, minipig and rabbit fibres in a single muscle section stained with this technique are identical to those obtained with the usual method based on a comparison of serial sections of which one is stained for SDH activity the other for reversed-ATPase activity.     

Zur funktionellen Bedeutung der osmiophilen Granula in Herzorganen niederer Vertebraten

Zusammenfassung Elektronenmikroskopische Untersuchungen am Herzen niederer Vertebraten (Frosch, Forelle, Goldfisch) und am Lymphherzen des Frosches zeigen, daß osmiophile intrazelluläre Granula (800–1300 Å) als Ausdruck sekretorischer Funktionen der Herzmuskelzellen bei verschiedenen poikilothermen Vertebraten vorkommen. — Innerhalb der Granulaansammlungen in Muskelzellen des Froschventrikels werden neben Dictyosomen, Vesikeln und Multivesikulärkörpern besondere Multivesikulärkörper mit Dichtekern gefunden. Übereinstimmende vesikuläre Substruktur dieses Dichtekerns und der spezifischen Granula sowie der Übergangsformen ergeben eine morphogenetische Reihe: Golgiapparat Multivesikulärkörper Multivesikulärkörper mit Dichtekern spezifisches Granulum. — Zwischen den Muskelzellen des Rana-Herzens wird wie bei Cyclostomen ein Zelltyp mit osmiophilen Granula gefunden, die in Größe und Struktur auch den Granula in den terminalen adrenergen Nervenfasern des Froschherzens entsprechen.Vieltägige Isolation des Froschherzens unter physiologischen Bedingungen sowie Inkubierung oder längere Behandlung isolierter Forschherzen mit Reserpin führt zum Abbau der spezifischen Granula. Oxypertin und Acetylcholin haben nur geringen Einfluß auf deren Größe, Struktur und Zahl. L-Dopa und Adrenalin wirken einer isolationsbedingten Reduktion der Granula entgegen. — Fluoreszenzmikroskopisch kann am Rana-Herzen intramuskulär eine auf Catecholamine hindeutende punktartige Gelbgrün-Fluoreszenz nachgewiesen werden. Nach Rückgang des Granulagehaltes durch Reserpin zeigt das isolierte Froschherz eine Sensibilisierung für Acetylcholin und Adrenalin.Es ist wahrscheinlich, daß die spezifischen Granula in den Herzorganen der untersuchten Arten (wie die entsprechenden Strukturen im Cyclostomenherzen) Catecholamine speichern, die für eine gleichbleibende Erregbarkeit der Muskelfasern von Bedeutung sein könnten.

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Function of the osmiophilic granules in the heart-organs of lower vertebratesA comparative electron microscopic and pharmacological study on the isolated heart of frog and other poikilothermic vertebrates

Summary Electron microscopic studies of the hearts of lower vertebrates (frog, trout, goldfish) and of the lymph-heart (frog) show that osmiophilic intracellular granules probably are related to secretory functions of cardiac muscle cells in poikilothermic vertebrates. — Beside dictyosomes, vesicles, and usual multivesicular bodies, a special type of multivesicular body with a dense core is found among the granules in the muscle cells of the frog's ventricle. The vesicular substructure of this dense core, and the specific granules as well as the intermediate forms suggest the following morphogenetic process: Golgisystem multivesicular body multivesicular body with a dense core specific granule. Between the muscle cells of the heart of Rana, as well as with Cyclostomata, a cell type with osmiophilic granules can be found corresponding in size and structure to the adrenergic terminals of the frog's heart.Many days' isolation of the frog's heart under physiological conditions, as well as incubation, or longer treatment of isolated hearts of frogs with reserpine leads to the dissolution of the specific granules. Oxypertin and acetylcholine have only little influence on the size, structure, and number of granules. L-Dopa and adrenalin appose a reduction of granules caused by isolation. Fluorescence microscopic results show a point-like yellow-green fluorescence in the heart of Rana, pointing to an accumulation of catecholamines intramuscularly.— After reduction of the content of granules by reserpine, the isolated frog's heart shows a higher sensitivity to acetylcholine and adrenalin.It is, therefore, probable that the specific granules store catecholamines in the heart organs of the investigated species in the same way as corresponding structures do in the hearts of Cyclostomata. These catecholamines are possibly important for a continual irritability of the cardiac muscle fibres in poikilothermic vertebrates.

Frau A. Beyerle-v. Wehren sei für ihre wertvolle technische Unterstützung gedankt.     

Myosatellite cells associated with different muscle fibre types in the Atlantic hagfish (Myxine glutinosa,L.)

Summary The incidence of myosatellite cells associated with white and red muscle fibres of the parietal muscle and red fibres of the craniovelar muscle was estimated by quantitative electron microscopy in the Atlantic hagfish (Myxine glutinosa, L.). Myosatellite cell nuclei constitute 3, 11 and 23 % of the total number of nuclei inside the basal lamina of the three types of muscle fibres, respectively. However, the total number of nuclei is highest in white fibres, most of the nuclei belonging to striated muscle cells. Myosatellite cell profiles in transverse sections constitute 23, 41 and 61 % of the number of muscle fibre profiles of the three types, respectively. The intervals between adjacent myosatellite cells are 135 m in white fibres, 55 m in red parietal fibres, and only 25 m in craniovelar fibres. Since craniovelar fibres are also comparatively thin, myosatellite cells constitute a significant fraction of the volume inside the basal lamina in these fibres. The myosatellite cells are 30–50 m long and up to 5 m thick. Some myosatellite cells possess few organelles, whereas others appear to contain many free ribosomes, granular endoplasmic reticulum, prominent Golgi apparatus and lysosome-like bodies.This investigation was supported by the Norwegian Research Council for Science and the Humanities (NAVF grant No. C20.30–37). The authors are indebted to Jorunn Line Vaaland and Berit Branil for technical assistance, and to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supplying the hagfish     

Zwei Muskelrezeptoren an der Mandibel von Leuctra (Insecta,Plecoptera) — Beispiele für nicht-ciliäre Sinnesorgane mit Tubularkörper-ähnlichen Strukturen

Zusammenfassung Beide Muskelrezeptoren an der Mandibel von Leuctra ziehen vom vorderen Tentorium-Arm zur Mandibel-Basis. Der ventrale Rezeptor besteht aus zwei dünnen Muskelfasern (6 bis 7 m Durchmesser) und mindestens 10 multiterminalen Sinneszellen, deren Dendriten sich im Innern der Fasern verzweigen und an den Z-Scheiben enden. Der dorsale Rezeptor besitzt drei ähnlich dünne Muskelfasern, aber nur eine einzelne multiterminale Sinneszelle. Ihre Dendriten enden im Ansatzgebiet des Muskels, zwischen Muskelfasern und Epidermiszellen.Beide Rezeptoren haben im wesentlichen denselben Feinbau wie bei Coleopteren, jedoch treten — besonders beim dorsalen Rezeptor — in den Dendriten-Enden Strukturen auf, die den Tubularkörpern bei Mechanosensillen ähneln.

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Two muscle receptor organs at the mandible of Leuctra (Insecta, Plecoptera) — examples for non-ciliary sense organs with tubular-body-like structures

Summary The muscle receptor organs of the mandible of Leuctra extend between the anterior tentorial arm and the mandible base. The ventral receptor is composed of two thin muscle fibres (6–7 m in diameter) and at least ten multiterminal sensory cells, the dendrites of which branch in the interior of the fibres and end near the z-bands. The dorsal receptor organ consists of three muscle fibres of similar diameter and only one multiterminal sensory cell. The dendritic ends lie at the distal end of the muscle, where muscle fibres and epidermal cells make contact.Both receptor organs essentially show the same ultrastructural characteristics as in Coleoptera. However, the dorsal receptor organ in particular possesses organelles in its dendritic ends, which look like the tubular bodies in ciliary mechanoreceptors.

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Mit Unterstützung durch die Deutsche Forschungsgemeinschaft     

Exhaustive physical exercise and acid hydrolase activity in mouse skeletal muscle

Summary Adult, untrained NMRI mice were exhausted on a motor-driven treadmill by an intermittent-type running programme. Serial cryostate sections for the staining of NADH-tetrazolium reductase, -glucuronidase, -N-acetylglucosaminidase, and -glycerophosphatase activities and for making hematoxylin-eosin staining were cut from m. quadriceps femoris 1, 2, 3, 5, 7, and 15 days after physical exhaustion. A strong increase in the activities of -glucuronidase and -N-acetylglucosaminidase, was observed 7 days after exhaustion and the activity changes, which were similar for the both glycosidases, were more prominent in the highly oxidative red compared to less oxidative white fibres. Activity granules were more numerous in the perinuclear than the interfibrillar area of red fibres. Spots were arranged like longitudinal chains between myofibrils. Activity in connective tissue was usually observed only in animals exhausted 3–7 days earlier. Simultaneous activity in fibres exceeded that in connective tissue -Glycerophosphatase activity was not, by the method used, seen in histologically healthy or normal-looking fibres. in samples taken 2–5 days after exhaustion some degenerating and necrotic fibres were observed. Inflammatory reaction was also observed being at its strongest five days after loading when mononuclear cells were seen inside necrotic fibres. The number of regenerating muscle cells was most abundant 7 days after exhaustion. It is suggested that temporary hypoxia, which accompanies exhaustive physical exercise in skeletal muscle, upsets the energy metabolism and homeostasis of fibres and causes the observed histological and histochemical alterations, which posses features typical of both lethal and sublethal acute cell injury.     

The histochemical demonstration of myoglobin and succinic dehydrogenase activity in the tibialis anterior muscle of the rabbit

Summary The staining reactions for myoglobin and succinic dehydrogenase activity in the tibialis anterior of the rabbit demonstrate four types of muscle fibre. These may be distinguished by their intensity of staining for myoglobin and the distribution of the mitochondria shown by the dehydrogenase reaction.The large fibres (70–80 m diameter) which contain many mitochondria evenly scattered throughout the fibre contain much myoglobin. Smaller fibres (45–60 m diameter) which show an identical staining reaction for the dehydrogenase reaction contain less myoglobin. This suggests that myoglobin may be present to aid the diffusion of oxygen into muscle fibres.     

Genetic analysis of brown planthopper resistance in twenty varieties of rice,Oryza saliva L.

Summary The inheritance of resistance to brown planthopper, Nilaparvata lugens (Stol.), of 20 rice cultivars was studied. Single dominant genes that are allelic to Bph 3 condition the resistance in cultivars Ptb 19, Gangala (Acc. 7733), Gangala (Acc. 15207), Horana Mawee, Kuruhondarwala, Mudu Kiriyal and Muthumanikam. Single recessive genes that are allelic to bph 4 govern the resistance in cultivars Gambada Samba, Heenhoranamawee, Hotel Samba, Kahata Samba, Kalukuruwee, Lekam Samba, Senawee, Sulai, Thirissa and Vellai Illankali. The resistance in Ptb 33, Sudu Hondarwala, and Sinna Sivappu is governed by one dominant and one recessive gene which segregate independently of each other. The dominant resistance genes in these cultivars appear allelic to either Bph 1 or Bph 3. Similarly, the recessive genes in these cultivars seem allelic to either bph 2 or bph 4. Further investigations are needed to conclusively determine the allelic relationships of resistance genes in Ptb 33, Sudu Hondarwala and Sinna Sivappu.     

A second order mechanical model of muscle spindle primary endings

Summary With reference to experimental data and the failure of earlier proposed first order linear models of mammalian muscle spindles, a second order mechanical model of de-efferented primary endings is studied. The model takes into account the presence of two different types of intrafusal muscle fibres in a complete spindle organ. It further allows the incorporation of different gain of the mechano-electric conversion into a depolarization of the sensory terminals innervating the two types of fibres.It is shown that a closer approximation to the behaviour of the biological prototype is obtained if the transducer gain of the branch corresponding to the nuclear bag intrafusal fibres is chosen significantly higher than that corresponding to the nuclear chain branch.The marked nonlinear behaviour of muscle spindle primary endings as recently reported by Matthews and Stein (1968, 1969) is interpreted as a saturation effect of the high gain mechano-electric transducer of the nuclear bag branch. The saturation is considered to reflect a condition of complete depolarization of these sensory terminals. If a higher transducer gain actually is present, a complete depolarization of these terminals would occur at a lower degree of mechanical deformation than for the nuclear chain terminals. The mechano-electric transducer system of the nuclear chain fibres might thus behave approximately linearly within a larger range of input amplitudes. The greatly reduced gain of the primary endings at large emplitudes of imposed muscle vibrations as observed experimentally (Matthews and Stein, 1968, 1969) may thus be accounted for by the transducer gain of the nuclear chain fibres alone.List of Symbols Used C Viscous stiffness, or electrical capacitance - f Frequency of a signal - K Static gain of a system - k Elastic stiffness - R Electrical resistance - s The Laplace operator - H(s) General transfer function of a system - X (s) Laplace transform of the difference between instantanous and resting length of the complete intrafusal muscle fibre, according to the suggested model shown in Fig. 2 - (s) Laplace transform of the length of the elastic component of the proposed Maxwell branch of the sensory region of the model - (s) Laplace transform of the difference between instantanous and resting length of the lumped model of the polar (non-sensory) region of the intrafusal fibres - (s) Laplace transform of the length of the purely elastic branch of the model - The transducer gain of the output from the -branch relative to that of the -branch - v(s) Laplace transform of the total output signal (s) + (s) - Time constant defined by or =RC for the mechanical and the electrical system respectively - Angular frequency equal to 2f - Rate constant describing the relation between the lead and the lag time constant of a first order lead-lag filter network     

Histochemical properties and innervation pattern of fast and slow-tonic fibre types of the anterior latissimus dorsi muscle of the chick

Summary The anterior latissimus dorsi muscle of the chick is largely composed of slow-tonic fibres but contains a few fast fibres defined by their acid-labile, alkali-stable myofibrillar ATPase activity. These fibres are referred to as fibres. Differing from the slow-tonic ( and ) fibres which are multiply innervated withen grape endings, the (fast) fibres are innervated by typicalen plaque endings. The innervation of thirty-two fibres were studied closely and it was concluded that about a half of the fibres were focally innervated whereas a half were innervated in two distinct loci byen plaque endings. In only one case, a fast fibre with three widely spaceden plaque endings was observed. The mean intersynaptic length was significantly larger on fibres than on and fibres. No variation in the histochemical properties of myofibrillar ATPase was observed either along the entire length of singly and dually innervated fibres, or along and fibres. It is concluded that the three extrafusal fibre types of the anterior latissimus dorsi muscle maintain uniform histochemical characteristics along their length in spite of a possible innervation by several motoneurons.     

Untersuchungen an isolierten Muskelspindeln der Ratte nach Cholinesterasedarstellung und Sudanschwarz-Färbung

Zusammenfassung An isolierten Muskelspindeln aus Lumbrical-Muskeln der Ratte, deren motorische Endigungen mit einer Cholinesterase-Technik dargestellt, deren Nerven- und Muskelfasern mit Sudanschwarz gefärbt worden waren, wurden lichtmikroskopisch folgende Befunde erhoben: Es gibt 2 Typen intrafusaler Muskelfasern: a) helle, den nuclear bag fibres entsprechende, b) dunkle, den nuclear chain fibres entsprechende.An beiden Fasertypen kommen distinkte motorische Endplatten vor, die gleichfalls in zwei Typen differenzierbar sind: a) Größere, aus locker gefügten, runden Untereinheiten bestehende; diese befinden sich nur an den hellen Fasern. Ihre Anzahl an jedem Polabschnitt kann bis zu drei betragen. b) Kleinere, kompakter gebaute motorische Endplatten; diese liegen nur an den dunklen intrafusalen Fasern, je Faserpol maximal eine.Außer den mehr in Polmitte liegenden distinkten Endplatten sieht man in den paraäquatorialen Polabschnitten regelmäßig eine zweite Art intrafusaler motorischer Nervenendigungen: die multiterminalen motorischen Endigungen. Diese an jedem Spindelpol sichtbaren Endigungen sind vorwiegend an den hellen intrafusalen Fasern lokalisiert.

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Studies on isolated muscle spindles of the rat after cholinesterase and Sudan black stainingA contribution to the morphology and distribution of motor nerve endings in the muscle spindles of mammals

Summary After staining of the motor nerve endings with a cholinesterase technique and of nerve and muscle fibers with Sudanschwarz B, single muscle spindles were isolated from lumbrical muscles of rats and examined by means of light microscopy.Two types of intrafusal muscle fibres were found: a) pale ones corresponding to nuclear bag fibres and b) dark ones corresponding to nuclear chain fibres.On both types of muscle fibres distinct motor end plates were observed; two types of endplates can be distinguished: a) Larger ones consisting of circular subunits in loose arrangement; they occur on pale muscle fibres only. Their number per muscle fibre pole may amount up to three, b) Smaller ones with more compact arrangement of their subunits occurring on dark fibres only. Their number per muscle fibre pole never exceeds one.Besides these distinct endplates with more mid-polar positions, a second kind of motor nerve ending is to be found in the juxtaequatorial region of every spindle pole: the multiterminal motor ending. These endings are located predominantly on pale intrafusal muscle fibres.An often very thin -fibre leads to every juxtaequatorial region, looses its myelin sheath and divides into several thin branches; these non-myelinated axon-branches may be observed over rather long distances, finally forming the multiterminal endings.Every distinct motor endplate receives its own — usually somewhat thicker—-fibre which looses its myelin sheath immediately before its termination at the endplate. The terminal parts of these fibres show no branching.The dark intrafusal muscle fibres receiving one motor end plate only, correspond to those intrafusal fibres of the rat which, according to other authors, show properties of twitch fibres. The pale fibres receiving multiterminal endings and multiple motor endplates, correspond to fibres which show some criteria of slow fibres.

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Herrn Prof. Dr. med. E. Horstmann zum 60. Geburtstag gewidmet     

Development and parasitism in the genus Hemileia (uredinales)

Seventeen species ofHemileia were studied with emphasis on these characters: extent of colonization, specialization of parasitic mycelium, haustorial development, pathologic histology, soral morphology and sporulation capacity. An attempt was made to trace the varying degree of development and parasitism in this rust genus comprising of just over 40 reported species. It was noted that theGopalkrishnan's Subepidermal type more advanced than his Superstomal — B type.Fourth part of my thesis submitted to the University of Poona, India, accepted in 1968 for Ph.D. Not published.     

Genotype effects on plant regeneration in callus and suspension cultures of Avena

Regenerative potential of the calli of nineteen genotypes of Avena sativa, Avena nuda, Avena byzantina and one interspecific hybrid were compared over three successive cultures. Highly significant genotype and genotype × subculture interactions were observed. Among the highest plant regenerable genotypes were Corbit (first subculture); GAF/Park and 88Ab3073 (second subculture); and GAF/Park and 87Ab5932 (third subculture). These genotypes regenerated on an average 10 to 17 plants each from a 200 mg callus mass after a 30 to 45 proliferation period. GAF/Park, a progeny of an interspecific cross, regenerated plants at a significantly higher level (11.85 plants/rep), followed by the similarly performing A. sativa (6.23 plants) and A. nuda (5.06 plants) genotypes, which were significantly higher than the A. byzantina genotypes (2.07 plants). Four genotypes were tested for their adaptability to suspension culture and plant regeneration potential by separating their cells and cell clusters into two sizes: larger and smaller than 3 mm. Larger clusters yielded plants for three genotypes GAF/Park, 88Ab3073, and Tibor. The smaller clusters only regenerated plants for GAF/Park and 88Ab3073. From one gram of callus used to initiate suspensions of GAF/Park and 88Ab3073, 119.9 and 18.8 plants, respectively, were regenerated. The plants regenerated for various genotypes from agar-solidified or suspension culture experiments had normal growth and seed set. This study confirms high and sustained regenerative capabilities of GAF/Park, a restricted genotype due to the weedy Avena fatua genetic background and identifies alternative genotypes, especially 88Ab3073 for future tissue culture and transformation studies.     

Cholinesterases and biogenic monoamines in ganglion semilunare (Gasseri)

Summary Cholinesterase distribution in trigeminal ganglion was studied in rabbits by the thiocholine method. The presence of acetylcholinesterase conditions either a) a diffusive cytoplasmatic reaction of varying intensity — from very weak to very strong — or b) an unevenly intensively distributed reaction pronounced on the periphery of ganglion cells. — The histochemical localization of biogenic monoamines was studied with the aid of the fluorescent method according to Falck and Hillarp. Green-yellow varicose terminals were found in the neighbourhood of blood vessels, between myelinized fibres, and, finally, in close vicinity of ganglion cells with which they probably form synaptic contacts.Preliminary paper read at the XVIIth Days of Physiology in Brno (Luká, Buriánek, ech 1966).     

Studies on adenosine triphosphate transphosphorylases. XVIII. Synthesis and preparation of peptides and peptide fragments of rabbit muscle ATP-AMP transphosphorylase (adenylate kinase) and their nucleotide-binding properties

Two peptide fragments, derived from the head and tail of rabbit muscle myokinase, were found to possess remarkable and specific ligand-binding properties (Hamadaet al., 1979).By initiating systematic syntheses and measurements of equilibrium substrate-binding properties of these two sets of peptides, or portions thereof, which encompass the binding sites for (a) the magnesium complexes of the nucleotide substrates (MgATP^2– and MgADP^–) and (b) the uncomplexed nucleotide substrates (ADP^3– and AMP^2–) of rabbit muscle myokinase, some of the requirements for binding of the substrates to ATP-AMP transphosphorylase are being deduced and chemically outlined. One requirement for tight nucleotide binding appears to be a minimum peptide length of 15–25 residues. In addition, Lys-172 and/or Lys-194 may be involved in the binding of AMP.The syntheses are described as a set of peptides corresponding to residues 31–45, 20–45, 5–45, and 1–45, and a set of peptides corresponding to residues 178–192, 178–194, and 172–194 of rabbit muscle adenylate kinase. The ligand-binding properties of the first set of synthetic peptides to the fluorescent ligands: MgATP/ATP and MgADP/ADP are quantitatively presented in terms of their intrinsic dissociation constants (K_d) and values ofN (maximal number of moles bound per mole of peptide); and compared with the peptide fragment MT-I (1–44) obtained from rabbit muscle myokinase (Kubyet al., 1984) and with the native enzyme (Hamadaet al., 1979). In addition, the values ofN andK_d are given for the second set of synthetic peptides to the fluorescent ligands AMP and ADP as well as for the peptide fragments MT-XII(172–194) and CB-VI(126–194) (Kuby et al., 1984) and, in turn, compared with the native enzyme.A few miscellaneous dissociation constants which had been derived kinetically are also given for comparison (e.g., theK _i for AMP and the value of obtained for the native enzyme) (Hamada and Kuby, 1978), and theK'_d measured for Cr³⁺ and the synthetic peptide I_1–45 (Fryet al., 1985b).Paper XVII of this series is Kubyet al. (1983).     

The histochemical specificity of High Iron Diamine—Alcian Blue

Summary The specificity of the High Iron Diamine—Alcian Blue pH2.5 (HID—AB 2.5) procedure was examined in tissue sites containing sialogycoproteins alone or differing proportions of sialo- and sulphosialoglycoproteins. Studies with HID in differing final concentrations of hydrochloric acid or sodium chloride confirmed that staining is dependent upon both the pH and the ionic strength of the dye bath and demonstrated a marked heterogeneity in the pKa of the anionic groups of sialosulphoglycoproteins. Use of the sequence High Iron Diamine—Alcian Blue pH 1.0 demonstrated that complete or almost complete staining ofO-sulphate esters occurred when HID was prepared in water (final pH 1.3). However, under these conditions HID—AB 2.5 was shown to be non-specific because only black HID staining was observed in sites containing large quantities of sialic acids. This non-specificity was due either to the masking of Alcian Blue staining by HID and/or the black HID staining of anionic groups other than sulphate. These results may account for some of the conflicting data obtained by different groups of investigators who have studied transitional mucosa in human colonic diseases. Caution should be used in drawing conclusions from the use of HID—AB 2.5 without confirmatory evidence from other more specific procedures.