Interactions between intrinsic regulation and neural modulation of acetylcholinesterase in fast and slow skeletal muscles

1. Initiation of subsynaptic sarcolemmal specialization and expression of different molecular forms of AChE were studied in fast extensor digitorum longus (EDL) and slow soleus (SOL) muscle of the rat under different experimental conditions in order to understand better the interplay of neural influences with intrinsic regulatory mechanisms of muscle cells. 2. Former junctional sarcolemma still accumulated AChE and continued to differentiate morphologically for at least 3 weeks after early postnatal denervation of EDL and SOL muscles. In noninnervated regenerating muscles, postsynaptic-like sarcolemmal specializations with AChE appeared (a) in the former junctional region, possibly induced by a substance in the former junctional basal lamina, and (b) in circumscribed areas along the whole length of myotubes. Therefore, the muscle cells seem to be able to produce a postsynaptic organization guiding substance, located in the basal lamina. The nerve may enhance the production or accumulation of this substance at the site of the future motor end plate. 3. Significant differences in the patterns of AChE molecular forms in EDL and SOL muscles arise between day 4 and day 10 after birth. The developmental process of downregulation of the asymmetric AChE forms, eliminating them extrajunctionally in the EDL, is less efficient in the SOL. The presence of these AChE forms in the extrajunctional regions of the SOL correlates with the ability to accumulate AChE in myotendinous junctions. The typical distribution of the asymmetric AChE forms in the EDL and SOL is maintained for at least 3 weeks after muscle denervation. 4. Different patterns of AChE molecular forms were observed in noninnervated EDL and SOL muscles regenerating in situ. In innervated regenerates, patterns of AChE molecular forms typical for mature muscles were instituted during the first week after reinnervation. 5. These results are consistent with the hypothesis that intrinsic differences between slow and fast muscle fibers, concerning the response of their AChE regulating mechanism to neural influences, may contribute to different AChE expression in fast and slow muscles, in addition to the influence of different stimulation patterns.     

Increased Acetylcholine Synthesis and Release in Brains of Cats with GM1 Gangliosidosis

Cholinergic processes were measured in motor cortex, hippocampus, and striatum of cats in the terminal stages of GM1 gangliosidosis and compared to those of control cats. The greatest difference observed was elevation in the rate of K+-stimulated release of acetylcholine (ACh) from brain slices prepared from affected cats. The K+-stimulated release of endogenous ACh was increased by 31-43% and of newly synthesized ACh by 19-80% in brain slices from different brain regions. All regions that were examined were affected but the greatest effects occurred in cortex. The rate of synthesis of ACh was elevated in cortical and hippocampal slices. Choline acetyltransferase activity in brain regions of cats with GM1 gangliosidosis was not significantly different from that in controls, whereas high-affinity choline transport in cortical synaptosomes was elevated. Muscarinic receptor binding sites were reduced in the cortex, hippocampus, and striatum of GM1 mutant cats, whereas the apparent affinity was not altered. These results indicate that there are major alterations of cholinergic function in the brains of cats with GM1 gangliosidosis.     

Temporalis and masseter muscle function during incision in macaques and humans

Most previously published electromyographic (EMG) studies have indicated that the temporalis muscles in humans become almost electrically quiet during incisai biting. These data have led various workers to conclude that these muscles may contribute little to the incisai bite force. The feeding behavior and comparative anatomy of the incisors and temporalis muscles of certain catarrhine primates, however, suggest that the temporalis muscle is an important and powerful contributor to the bite force during incision. One purpose of this study is to analyze the EMG activity of the masseter and temporalis muscles in both humans and macaques with the intention of focusing on the conflict between published EMG data on humans and inferences of muscle function based on the comparative anatomy and behavior of catarrhine primates. The EMG data collected from humans in the present study indicate that, in five of seven subjects, the masseter,anterior temporalis, and posterior temporalis muscles are very active during apple incision (i.e., relative to EMG activity levels during apple and almond mastication). In the other two human subjects the EMG levels of these muscles are lower during incision than during mastication, but in no instance are these muscles ever close to becoming electrically quiet. The EMG data on macaques indicate that, in all six subjects, the masseter, anterior temporalis, and posterior temporalis muscles are very active during incision. These data are in general agreement with inferences on muscle function that have been drawn from the comparative anatomy and behavior of primates, but they do not agree with previous experimental data. The reason for this disagreement is probably due to differences in the experimental procedure. In previous studies subjects simply bit isometrically on their incisors and the resulting EMG pattern was compared to the pattern associated with powerful clenching in centric occlusion. In the present study the subjects incised into actual food objects, and the resulting EMG pattern was compared to the pattern associated with mastication of various foods. It is not surprising that these two procedures result in markedly different EMG patterns, which in turn result in markedly different interpretations of jaw-muscle function. In an attempt to explain the evolution of the postorbital septum in anthropoids, it has been suggested that the anterior temporalis is more active than the masseter during incision (Cachel, 1979). The human and macaque EMG data do not support this hypothesis; during incision, the two muscles show no consistent differences in humans and the masseter appears to be in fact more active than the anterior temporalis in macaques.     

Implementation of periodic acid-thiosemicarbazide-silver proteinate staining for ultrastructural assessment of muscle glycogen utilization during exercise

Summary Distribution of glycogen particles in semithin and ultrathin sections of biopsy samples from human muscles subjected to either short- or long-term running were investigated using PAS and Periodic Acid-ThioSemiCarbazide-Silver Proteinate (PA-TSC-SP) staining methods. Glycogen particles were predominantly found immediately under the sarcolemma or aligned along the myofibrillar Iband. After long-term exhaustive exercise type-1 fibers with a few or no glycogen particles in the core of the fibers were frequently observed. The subsarcolemmal glycogen stores of these depleted type-1 fibers were about three times as large as after exhaustive short-time exercise. Another indication of utilization of subsarcolemmal glycogen stores during anaerobic exercise was that many particles displayed a pale, rudimentary shape. This observation suggests fragmental metabolization of glycogen. Thus, depending on type of exercise and type of fiber differential and sequential glycogen utilization patterns can be observed.     

Utilisation de lois de la mécanique pour l'Estimationde la vitesse de locomotion des Vertébrés tétrapodes du passé

Two computation methods are explained; theirobject is the estimation of the velocity of animals which are known by their footprints, and in the case of the first method, by their skeletons as well.The first method is based on the compound pendulum theory, because during the slow walking gait, the motion of the leg is similar to the oscillation of a pendulum, for the computation of the velocity (v), are considered: moment of inertia (I), radius of gyration (P) and period (T), time in seconds to cover one stride (E) in the case the maximum angle of divarication (δ) of the leg with the vertical is ≤20°. A comparison with the formula of Alexander (1976) is discussed.The second method concerns saltator animals. It is based on the fondamental laws of dynamics. With the length of the jump (E) it is possible to estimate the velocity of the trackmaker (v) and the height of the jump (K). For the vertebrates the angle of the trajectory with the horizontal plan (α) is between 20 and 45°. Thus, the result of this method is an interval of estimation in which the velocity is included.These methods do not give precise results. But these approximations supply valid informations on the velocity of extinct or living animals and can lead to the estimation of their maximum speed if the parameters E, T, vary. The problem of the errors is also discussed. It is shown that the errors in estimating the parameters have a no significant influence on the results.     

An ultrastructural investigation of muscle attachment in the opercular filament of a polychaete annelid

The ultrastructure of peduncle muscle attachment to the cuticular flange in the opercular filament of the serpulid Pomatoceros lamarckii Quatrefages is described. The cuticular flange is composed of layers of orthogonally arranged fibres. Specialized epithelial cells (tendon cells) and a collagenous matrix intervene between the peduncle muscles and the cuticular flange. The tendon cells are characterized by hemidesmosomes at both apical and basal ends, connected by thick bundles of tonofilaments. Apically long specialized microvilli from the tendon cells penetrate the cavities in the orthogonally arranged layers of fibres of the cuticular flange. The basal surfaces of the tendon cells and the terminal ends of the peduncle muscles anchor independently of one another in the collagenous matrix. The peduncle muscles appear to be smooth muscles which contain thin filaments, 5 nm in diameter, and thick filaments, 40-100 nm in diameter, with a faint axial periodicity 12-14 nm. The method of peduncle muscle attachment in the opercular filament is compared with those of other invertebrates.     

Expression of creatine kinase isoenzyme during oogenesis and embryogenesis in the mouse

Creatine kinase activity was discovered in the growing mouse oocyte and in the preimplantation embryo. Changes in the enzyme activity during the growth and maturation of the egg and during the development of the embryo up to the blastocyst stage were determined. Close similarity of the protein to the brain-type isoenzyme of creatine kinase was established immunochemically. The kinetic parameters of the brain-type isoenzyme (M. R. Iyengar, C. E. Fluellen, and C. W. L. Iyengar, 1982, J. Muscle Cell Motil. 3, 231–246) and the pattern of development-associated changes in activity suggest a possible role for creatine kinase in maintaining the reported high ATP/ADP ratio (L. Ginsberg and N. Hillman, 1975, J. Reprod. Fertil. 43, 83–90), which is essential for the biosynthetic activities of the embryo.     

Zur Funktion der Mikrotubuli beim Wachstum der Myofibrillen von Insektenflugmuskeln

Zusammenfassung In der indirekten Flugmuskulatur von Phormia terrae-novae kann die Entwicklung der Myofibrillen deutlich in eine Anlage- und eine Wachstums-Phase unterteilt werden. Zu Beginn der Wachstumsphase wurde Puppen eine Lösung von Colchicin in die rechte Metathoraxhälfte injiziert. Als Folge dieser Behandlung lösten sich zunächst die Mikrotubuli in der Flugmuskulatur auf. In späteren Entwicklungsstadien bildeten sich atypische Verzweigungen der Myofibrillen, die zu einer partiellen Desorientierung der kontraktilen Strukturen führten. Ein Mechanismus, der solche Störungen des Orientierungmusters in der Normalentwicklung möglicherweise verhindert, wird diskutiert.

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On the function of microtubules during the growth period of myofibrils in insect flight muscles

Summary In the blowfly Phormia terrae-novae the development of myofibrils of indirect flight muscles can be divided into periods of predisposition (anlage) and of growth. At the beginning of the growth period microtubules are disrupted by injection of colchicine. This disruption is followed by the formation of atypical ramifications of myofibrils at Z-discs leading to numerous disoriented myofibrils in late developmental stages. A possible mechanism preventing these alterations during normal development is discussed.

Herrn Prof. Dr. H. Brettschneider bin ich für den in großzügiger Weise überlassenen Arbeitsplatz am Elektronenmikroskop zu Dank verpflichtet. Fräulein H. Bock danke ich für ausgezeichnete Hilfe.     

Unusual responses of proboscis muscles ofBusycon canaliculatum to some calcium antagonist agents

Summary K- and ACh-induced responses of the radular sac, odontophore retractor, and radular retractor muscles ofBusycon canaliculatum were found to be strongly dependent upon [Ca]₀. Diltiazem had strong positive inotropic and chronotropic actions on fast twitch activity in the odontophore retractor and radular protractor muscles. K-induced tonic force in these muscles was partly inhibited by diltiazem but only at very high concentrations. ACh responses in all muscles were eliminated by diltiazem. Nifedipine enhanced fast twitches and tonic force in response to high K, and induced persistent spontaneous fast twitch discharges. Nifedipine inhibited ACh-induced tonic force, but induced rhythmic bursts of fast twitches persisting long after nifedipine washout. Verapamil strongly inhibited K- and ACh-induced tonic force in all three muscles at high concentration, but stimulated fast twitch responses and converted ACh contractures into fast twitch activity. Sucrose gap studies showed that nifedipine and diltiazem reduced K- and ACh-induced tension and depolarization. Paradoxically, verapamil reduced K- and ACh-induced tension but significantly enhanced their induced depolarizations. Diltiazem, nifedipine and verapamil did not act like slow Ca channel antagonists in these muscles. This may reflect differences in channel structure between molluscs and mammals, or differences in the cellular calcium release pathways operated by such channels in molluscan and mammalian muscle. These Ca-ant-agonists appeared to act as agonists of fast twitch activity in these muscles and antagonists of the ACh-induced calcium release pathway for tonic force development.