Mechanical properties of frog muscle fibres at rest and during twitch contraction.

Data reported in the literature suggest that crossbridges in rapid equilibrium between attached and detached states (weakly binding bridges), demonstrated in relaxed skinned fibres at low ionic strength, could be present also in intact fibres under physiological conditions. In addition, it was suggested that the well known leading of stiffness over force during the tension development in stimulated muscle fibres could be due to an increased number of weakly binding bridges induced by the stimulation. The experiments reviewed in this paper were made to investigate these possibilities. Fast ramp length changes were applied to single frog muscle fibres at rest and during the early phases of activation. The corresponding force changes were analysed, searching for the components expected from the presence of weakly binding bridges. The results showed no mechanical indication for the presence of weakly binding bridges in both skinned and intact fibres, either at rest or during activation. It was also found that a portion of the fibre stiffness increase induced by stimulation leads the formation of crossbridges.     

"Red" fibres of pigeon pectoralis major muscle are "type II red".

On the basis of the histochemical activity of succinic dehydrogenase, only two fibre-types are distinguished in pigeon pectoralis major muscle. These are narrow "Red" and broad "White". The histochemical activity of myofibrillar ATPase was studied in these two distinct fibre-types. Both fibre-types showed high activity for the ATPase. "Red" fibres of pigeon pectoralis were not alkali-labile, at incubation pH 9.4, as were the "Type I" fibres of both avian and mammalian muscles. Again unlike "Type I" fibres, the "Red" fibres of pigeon pectoralis lacked the characteristic activation of acid-preincubated ATPase reaction. Pigeon pectoralis "Red" fibres are known to possess some characteristics of fast-twitch fibres (e.g. high fat, considerable phosphorylase, fibrillenstruktur myofibrillar arrangement, focal "en plaque" pattern of nerve endings). It is emphasized, therefore, that the pigeon pectoralis "Red" fibres are not equivalent to "Type I or slow-twitch", muscle fibres, but they are possibly "fast-twitch fatigue resistent or Type II Red" muscle fibres.     

Dynamics of lesions to the myocardium in experimental hyper- cholesterolemia

Dynamics of changes in the myocardium of rabbits subjected to food hypercholesterolemia lasting from 6 to 16 weeks was investigated. An intensification of coronary atheromatosis was proportional to the duration of the high-cholesterol diet. There were observed focal and growing in time damages of cardiomyocytes. They were sharply outlined in atherosclerotically changed coronary arteries. The following morphological and histochemical changes have been observed: increased acidophilia and fuchsinophilia in the single and grouped muscle fibres, foci of infiltrations by mononuclear cells, contraction bands, and outlines of myocardial fibres, separation of myofibrils, granular disintegration of cardiomyocytes, healed infarcts, depletion and excessive accumulation of glycogen in muscle fibres, presence of neutral fats, cholesterol and its esters in atheromatous plaques of coronary arteries, presence of neutral fats in some cardiomyocytes: focal acid phosphates, loss of activity of Mg- and Ca-dependent ATPases and SDH: oedema of mitochondria with disorganization of cristae, disorganisation of fibres and lysis of myofilaments, margination of chromatin in cardiomyocyte nuclei, increased number of lysosomes, intensified symptoms of egzocytosis, widening of channels of sarcoplasmatic reticulum, oedema of endothelium of capillary vessels and increased number of the collagenous fibres in the interstitial space. The results of histological, histochemical and microscopic-electron investigations correlated with each other. It may be considered that the observed damages of cardiomyocytes precede myocardial infarction and result from progressive and increasing ischemia, hypoxia and accumulation of fat substances and cholesterol and its esters in intima of coronary arterioles as well as in cardiomyocytes.     

Length dependent state of activation — Length change dependent kinetics of cross bridges in skinned insect flight muscle

Stretch induced activation and release induced deactivation of single glycerol-extracted insect flight muscle fibres were investigated. The results are interpreted to indicate that the muscle length controls the number of acting cross bridges, whereas their attachment-detachment kinetics in mainly determined by the state of strain of the cross bridges. It is concluded that the net detachment rate of the cross bridges is enhanced if the muscle is released thereby “unloading” the cross bridges. This behaviour of the unloaded cross bridge is a basic postulation of most of the molecular muscle contraction models.

1. The delayed tension rise induced by stretches of different amplitudes could be restored to the level before the stretch by a release to the initial length.
2. The delayed tension decrease induced by a release of moderate (up to δL=1.5% L _i)amplitude is quantitatively restored within the delayed increase induced by the restretch to the initial length.
3. Stiffness, which decreased during the delayed tension drop after release, is restored during a delayed stiffness increase effected by a restretch to the initial length.
4. The rate and the extent of the stiffness drop after release increased with increasing amplitude of the release and with increasing temperature.
5. After the deactivation, i.e., after tension and stiffness achieved a new steady level after the release, the attached cross bridges are already in the same state of strain as they were before the release. This finding is interpreted to indicate that within the deactivation phase all cross bridges attached prior the release are replaced by cross bridges attached after the release.
6. The rate of tension and stiffness decay after release does not depend on the absolute muscle length but on the amplitude of the release which induced the deactivation.

     

Cross bridge slippage in skinned frog muscle fibres

Mechanically skinned single fibres of the semitendinosus muscles of Rana esculenta were investigated at ca. 4 degrees C. The fibres were activated by a Ca2+ jump technique, which allowed the development of a steady isometric tension within several seconds of entering a calcium rich solution at 4 degrees C. Sequences of length changes of different duration and amplitude were applied to the fibre. It could be demonstrated that the fibre behaved as a Hookean spring in the case of small amplitude length changes (up to 0.5% L0, ramp duration 0.5 ms) and that a sequence of length changes induced reversible changes in fibre state. In contrast, large stretches (greater than 1% L0) induced a muscle "give" if the stretch were not immediately preceded by a release. The data was interpreted on the basis of a strain induced detachment of cross bridges in combination with a rapid reattachment of presumably the same cross bridges in a discharged position. The rates of strain induced detachment and reattachment depended on the stretch amplitude. At amplitudes exceeding 2% L0 the rates were estimated to be at least several thousands per second.     

The "all-or-none" law in skeletal muscle and nerve fibres

In 1905 the Cambridge physiologist Keith Lucas extended the "all-or-none" principle (introduced by H. P. Bowditch for the cardiac tissue) to skeletal muscle and nerve fibres. Nevertheless, in a short time it was clear that nerve fibres obey this law, but also that frequency of discharge is another relevant factor in the nervous conduction.     

Twitchin of mollusc smooth muscles can induce “catch”-like properties in human skeletal muscle: support for the assumption that the “catch” state involves twitchin linkages between myofilaments

Molluscan catch muscles can maintain tension with low or even no energy utilization, and therefore, they represent ideal models for studying energy-saving holding states. For many decades it was assumed that catch is due to a simple slowing of the force-generating myosin head cross-bridge cycles. However, recently evidences increased suggesting that catch is rather caused by passive structures linking the myofilaments in a phosphorylation-dependent manner. One possible linkage structure is the titin-like thick filament protein twitchin, which could form bridges to the thin filaments. Twitchin is known to regulate the catch state depending on its phosphorylation state. Here, we found that twitchin induces a catch-like stiffness in skinned human skeletal muscle fibres, when these fibres are exposed to this protein. Subsequent phosphorylation of twitchin reduces the stiffness. These findings support the assumption that catch of molluscan smooth muscle involves twitchin linkages between thick and thin filaments.     

Morphometry, ultrastructure, myosin isoforms, and metabolic capacities of the "mini muscles" favoured by selection for high activity in house mice

Prolonged selective breeding of mice (Mus musculus) for high levels of voluntary wheel running has favoured an unusual phenotype ("mini muscles"), apparently caused by a single Mendelian recessive allele, in which most hind-limb muscles are markedly reduced in mass, but have increased mass-specific activities of mitochondrial enzymes. We examined whether these changes reflect changes in fibre size, number or ultrastructure in normal and "mini-muscle" mice within the two (of four) selectively bred lines (lab designations L3 and L6) that exhibit the phenotype at generations 26 and 27. In both lines, the gastrocnemius and plantaris muscles are smaller in mass (by >50% and 20%, respectively) in affected individuals. The mass-specific activities of mitochondrial enzymes in the gastrocnemius and plantaris muscles were increased in the mini phenotype in both lines, with stronger effects in the gastrocnemius muscle. In the gastrocnemius, the % myosin heavy chain (MHC) IIb was reduced by 50% in L3 and by 30% in L6, whereas the % MHC IIa and I were higher, particularly in L3. Fibre number in the plantaris muscle did not significantly differ between mini and normal muscles, although muscle mass was a significant positive correlate of fibre number. Small fibres were more abundant in mini than normal muscles in L3. Mitochondrial volume density was significantly higher in mini than normal muscle fibres in L3, but not in L6. Microscopy revealed a surprising attribute of the mini muscles: an abundance of small, minimally differentiated, myofibril-containing cells positioned in a disorderly fashion, particularly in the surface layer. We hypothesise that these unusual cells may be satellite cells or type IIb fibres that did not complete their differentiation. Together, these observations suggest that mice with the mini phenotype have reduced numbers of type IIb fibres in many of their hind-limb muscles, leading to a decrease in mass and an increase in mass-specific aerobic capacity in muscles that typically have a high proportion of type IIb fibres. Moreover, the several statistically significant interactions between muscle phenotype and line indicate that the effect of the underlying allele is altered by genetic background.     

Relationship between myoglobin and succinate dehydrogenase in mouse soleus and plantaris muscle fibres

Summary This report describes a quantitative histochemical study of myoglobin in skeletal muscle fibres. The muscle fibres were classified as fast or slow on the basis of their quantitative myofibrillar ATPase histochemistry. A large range of myoglobin absorbance values was found among fast skeletal muscle fibres. This range was relatively small among slow fibres. The concentrations of myoglobin and the activities of succinate dehydrogenase in individual muscle fibres in serial sections are weakly correlated in both the mouse soleus and plantaris muscle. The myoglobin concentration is higher in fast and slow oxidative soleus muscle fibres and the succinate dehydrogenase activity in these fibres is lower than in oxidative plantaris muscle fibres in the same range of cross-sectional area.     

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

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.     

Isometric contractions of the myocardium: mathematic modeling

A mathematical model describing a single contraction of a cardiac muscle strip under isometric conditions is proposed. The adequacy of the model was checked in experiments on cardiac strips from patients with chronic coronary insufficiency. It was shown that the contraction-relaxation cycle is rather completely characterized by the parameters characterizing the association-dissociation kinetics of actomyosin bridges.     

Autoregulation of the coronary vascular resistance after beta-adrenergic receptor blockade

It was previously observed that at the beginning of a 10 sec multiple arterial haemorrhage, vascular elasticity induces an increase of coronary vascular resistance, which in the late part of the haemorrhage is counteracted by the relaxation of the vascular smooth muscle fibres. In the present study the effect of multiple arterial haemorrhage on the coronary vascular resistance was studied after beta-adrenergic receptor blockade. The results suggest that also in these circumstances an elastic mechanism precedes a myogenic mechanism in the regulation of coronary vascular resistance.     

Histochemical and immunohistochemical investigation of muscle fibres in the sturgeon (Chondrostei; Acipenser)

The arrangement and innervation of lateral muscle and adductor mandibulae (AM) complex were examined histochemically and immunohistochemically in the sturgeon Acipenser transmontanus. Lateral muscle was arranged in myotomes in which the superficial slow-red muscle layer and the deep fast-white muscle layer were separated by an intermediate layer of pink muscle. The AM complex was composed of a small slow-red muscle layer, but the bulk of the muscle was composed of fast-white fibres. Pink fibres appeared both in an intermediate layer and scattered throughout the fast-white muscle. The innervation appeared to resemble that of elasmobranchs and some teleosts. Slow-red fibres of lateral muscle were multiply innervated, whereas fast-white muscle fibres were focally innervated at the myoseptal end. Pink fibres of lateral muscle and fast-white fibres of AM were focally innervated in the mid-region of the fibres. There were no significant differences between 5 month old and adult sturgeons except for a much better developed intermediate (pink) layer surrounding the lateral line in the adult.     

Overrelaxation of muscle models in media with high concentrations of calcium ions and exposed to MgATP in "contraction-inducing" concentrations]

Cross-striation pattern and sarcomere length in isolated myofibrils (both glycerinated and fresh) as well as isometric tension of glycerinated fibers of rabbit m. psoas are unaffected by an evaluation in ionic strength of CaCl2 up to 0.2 in the absence of ATP. An addition of MgATP (1 to 3mM) to the Ca2+ media induces the changes which have been shown to be characteristic of overrelaxation [1, 2]: A band shortening occurs followed by a complete plastification of the fibres. A tentative mechanism of the process is discussed in terms of spontaneous rearrangement of calcium myosinate packing in thick filaments that follows disrupting of rigor crossbridges with thin filaments under the action of ATP. Released calcium myosinate heads fail to form "active" bridges with actin; thick filaments undergo a conformational change resulted in their shattening due to increase in the equilibrium region of LMM tail overlap. The effects do not depend on ionic strength only: on replacing CaCl2 by KCl at equal ionic strength 0.2, an addition of ATP induces normal contraction instead of overrelaxation. A possibility is discussed that in a living muscle overrelaxation could provide a siding to prevent damage in case of emergency.     

Effect of in vivo cold-exposure on intracellular glycogen reserves in the "Starling type" avian pectoralis

The pectoralis muscle of the grackle ($\text{Quiscalus}$$\text{quiscula}$), as an example of the “Starling Type” of avian pectoralis whose fibre composition is characterized as consisting of “red” and “intermediate” fibres, was studied in order to determine the respective roles of the fibre types in shivering thermogenesis. Exposure of partially-defeathered grackles to cold (?25°C; 30 min) elicited an overt shivering-response in which a rapid depletion of the glycogen reserves in the intermediate fibres, but not in the red, was demonstrated histochemically. It is concluded that the intermediate fibres are chiefly, if not solely, responsible for effecting shivering thermogenesis in the grackle, as are the white fibres in the pigeon under indentical conditions of cold stress.     

The fine structure of muscle attachments in an acarid mite, Caloglyphus mycophagus (Megnin) (Acarina)

The fine structure of the myo-cuticular junction in an acarid mite, Caloglyphus mycophagus, is described. The muscle fibres are attached to the cuticle via flattened, much invaginated, epidermal cells. Unlike the situation described for other arthropods, the stress across these epidermal cells does not appear to be transmitted by microtubules but rather by desmosome-like structures which form intraepidermal cell bridges where invaginations from the outer and inner surfaces of the epidermal cells lie close together. The muscles are attached to the inner surface of this complex desmosome and the outer surface is linked to the cuticle by extracellular fibrils.     

Succinic dehydrogenase activity of the respiratory muscles of a fresh-water teleost, Bagarius bagarius (Ham.) (Sisoridae, Pisces).

Functional morphology including the origin, insertion, and innervation of the respiratory muscles in relation to buccal pressure pump and opercular suction pumps in a fresh-water bottom dwelling siluroid fish, Bagarius bagarius have been studied. Histochemical studies were made on the succinic dehydrogenase activity of adductor mandibulae, retractor tentaculi, levator operculi, dilatator operculi, adductor operculi, intermandibularis, interhyoideus, hyohyoideus superior and constrictor branchialis. The intensity of reaction reveals the presence of three types of muscle fibres in some of the respiratory muscles. The muscle containing red muscle fibres are mostly innervated by the branches of the VIIth cranial nerve. The retractor tentaculi consists of superficial white muscle fibres and the interior part is dominated by red muscle fibres. The muscles (adductor operculi, levator operculi, dilatator operculi, interhyoideus, hyohyoideus superior) concerned with the opercular suction pumps are of mixed type and consist of white and red muscle fibres, whereas adductor mandibulae and intermandibularis are made up entirely of white muscle fibres. The adductor muscle bundles of the constrictor branchialis, which are responsible for movement of gill filaments, are dominated by the red muscle fibres. The abductor part, however, is made up entirely of white muscle fibres.     

Simple physical model of coronary vasodilation

The coronary diastolic pressure-flow relationship was studied in two groups of dogs; in one group coronary circulation was characterized by normal tone of vascular smooth muscle, while in the other group, complete relaxation of smooth muscle fibres was produced by intravenous infusion of dipyridamole. The coronary flow (CF) was measured in both groups for several values of mean aortic blood pressure (ABP), the variations being obtained by means of a 10 s arterial haemorrhage. The measured CF versus ABP data were found to be well represented by best fit curves calculated by power regression methods. These curves were quite different in the presence and absence of dipyridamole infusion. A simple physical model is proposed for analysis of these curves; the model is based on the Poiseuille equation, modified to take into proper account the variations of the vessel radii under different ABP values during ventricular diastole. These variations are expressed by means of Laplace and Hooke's laws by equating wall tension due to APB, to the sum of tensions due to elastic and smooth muscle forces. Analysis of CF versus APB curves, performed on the basis of this model, shows that to account for the large change of coronary bed conductance observed under dipyridamole infusion one must assume not only that the smooth muscle tone is absent but also that some relevant variations occur in the whole coronary bed, thus pointing to possible recruitment of new blood vessel paths.     

Perfusion of the psoas muscle of the rabbit. Metabolism of a homogeneous muscle composed of "fast glycolytic" fibres

A method was developed for the hemoglobin-free perfusion of the rabbit psoas muscle in situ. This muscle consists almost exclusively of fast-twitch (type IIb) glycolytic fibres and was therefore used as a model of a homogeneous muscle of the glycolytic, metabolic type.     

The properties of the extraocular muscles of the frog. III. Morphological, mechanical and pharmacological properties of the isolated retractor bulbi muscle.

Some morphological, physiological, and pharmacological properties of the retractor bulbi muscle of the frog were tested. The enzyme-histochemical investigation shows that the retractor bulbi muscle contains twitch muscle fibres only. Two types of twitch muscle fibres, which are especially different in their diameter and in the content of mitochondria, build the muscle in an irregular arrangement; tonic muscle fibres were not observed. On the average, the isolated retractor bulbi muscle has at room temperature a contraction time of 26 ms, a half-relaxation time of 28 ms, a fusion frequency of 75 stimuli/s, and a twitch-tetanus ratio of 0.28. The fatigability of this muscle is higher than in oculorotatory eye muscles but lower than in skeletal muscles of the frog. An increase of the extracellular K+-concentration elicits in retractor bulbi muscles a quickly transient contracture; the mechanical threshold of the muscle fibres is found in a range between 20 and 25 mM K+ in Ringer solution. Similar short-lasting contractures, which are probably caused by twitch fibres, rich in mitochondria, are also evoked by application of depolarizing drugs like acetylcholine. The properties of the retractor bulbi muscle are compared with those of the sartorius muscle of the frog, which likewise contains twitch muscle fibres only.