Fiber composition of the distal accessory flexor muscle in several decapod crustaceans

The fiber composition of the distal accessory flexor muscle (DAFM) and the branching pattern of its excitor axon were compared in several species of crabs, in the lobster and the crayfish. The muscle is composed exclusively of long sarcomere (> 6 μm) fibers and therefore of the slow type. In all the crab species, except one, there is a distal to proximal gradient of fibers with increasing sarcomere lengths; this gradient is reverse in lobsters and crayfish. A proximal to distal gradient of increasing fiber diameters occurs in the DAFM of all crab species but not in the lobster and crayfish, in which all the fibers are approximately equal in diameter. The single excitatory axon traverses the width of the DAFM and gives off primary branches on either side in the lobster and crayfish but on only one side in crabs. The hypothesis that the axonal branching pattern may govern the regional distribution of fibers with differing sarcomere lengths in proposed.     

Parallel reflex and central control of promotor and receptor motoneurons in crayfish.

We describe the reflex and central control of an identified motoneuron (rm 1) to a crayfish muscle receptor, the thoracocoxal muscle receptor organ (TCMRO), and compare it with the in-parallel, 'extrafusal' promotor motorneurons. Rm 1 is spontaneously active in an isolated preparation. This activity is modulated in phase with centrally driven promoter nerve activity, suggesting coactivation of promotor and receptor-motor motoneurons. Rm 1 is autogenetically modulated, in a phase-dependent manner, by stretching the TCMRO: during promotor bursts rm 1 is excited by dynamic stretch, but during remotor bursts it is inhibited by the same stimulus. This effect is mediated by a single, identified TCMRO afferent, the dynamically sensitive T-fibre. At or near maximally stretched lengths of the TCMRO a tonic inhibition of rm 1 is revealed. This effect is mediated by another identified TCMRO afferent, the statistically sensitive S-fibre. The thoracocoxal chordotonal organ is a non-muscular receptor spanning the same joint but signalling the opposite direction of movement. Dynamic movement stimulation of this receptor also excites rm 1, a reflex that could counteract TCMRO slackening. These results demonstrate a complex central and reflex control of the TCMRO, which could regulate reflex gain throughout the step cycle during walking in the intact animal.     

Decrease in light diffraction intensity of contracting muscle fibres

Single fibres from the semitendinosus muscle of frog were illuminated normally with a He–Ne laser. The intensity transient and fine structure pattern of light diffracted from the fibre undergoing isometric twitches were measured. During fibre shortening, the intensity decreased rapidly and the fine structure pattern preserved its shape and moved swiftly away from the undiffracted laser beam. The fine structure patterns of the contracting and resting fibre were nearly identical. The ratio of intensities of the contracting and resting fibre of the same sarcomere length was determined as a function of the time elapsed after fibre stimulation. The time-resolved intensity ratio increased with sarcomere length and became unity when sarcomere length was between 3.5 m and 3.7 m. A diffraction theory based on the sarcomere unit was developed. It contained a parameter describing the strength of filament interaction. The comparison between the theory and data shows that the initial intensity drop during contraction is primarily due to filament interactions. At a later stage of contraction, sarcomere disorder becomes the major component causing the intensity to decrease. Diffraction models which use the Debye-Waller formalism to explain the intensity decrease are discussed. The sarcomere-unit diffraction model is applied to previously reported intensity measurements from active fibres.     

Distribution of calcium during contraction and relaxation of crayfish skeletal muscle fibre.

A model of activation of muscle contraction has been applied to the crayfish isolated skeletal muscle fibre. The model is based on calcium diffusion and binding to specific regulatory sites in a sarcomere. Calcium ions activate interactions of contractile proteins and thus the generation of force. The model quantifies the relation between calcium released from intracellular stores and force elicited. Experimental tension records from isolated crayfish skeletal muscle fibres under voltage clamp conditions are analyzed. Model parameters were determined either via approximation of the onset of tension by the model solution or from the model based relations between the tension maximum, and depolarizing pulse length and amplitude. This allowed to determine time changes of free and bound calcium distribution in the sarcomere and the calcium release from terminal cisternae. The steady state calcium concentration at terminal cisternae showed S-shaped voltage dependence with saturation below approx. 10 mumol/l at positive membrane potentials.     

Leverage and muscle type in crab chelae (Crustacea: Brachyura)

The chelae of Cancer pagurus and Macropipus depurator were examined with respect to mechanical advantage. The closer muscles were investigated with respect to sarcomere length in the constituent fibres and to the force developed by the whole muscle during isometric contraction. Cancer chelae have a relatively high mechanical advantage, 0.329 ± 001. Cancer closer muscles contain a high proportion of fibres with long sarcomeres, mean lengths mostly falling between 12 and 15 μm, and develop a maximum stress of about 496 kN.m⁻² during contraction. These figures are typical for "slow" crustacean muscle. The chelae of M. depurator are dimorphic. In one, the strong chela, the mechanical advantage is 0.248 ± 0.066 while in the other, the fast chela, the mechanical advantage is 0.177 ± 0.006. M. depurator closer muscles contain fibres with mean sarcomere lengths mostly falling between 6 and 10 μm. The muscle develops a maximum stress of about 145 kN.m⁻² during contraction. These figures are typical of "intermediate" crustacean muscles. "Fast" muscle fibres with short sarcomeres (about 30 um) were found in the chelae of both Cancer and M. depurator but were much commoner in the latter. Thus in Cancer a high mechanical advantage is correlated with slow muscle while in M. depurator lower mechanical advantages are broadly correlated with faster muscle. Consistent correlation between mechanical advantage and muscle type in the dimorphic chelae of M. depurator , however, is lacking. No consistent regionation of fibres with similar properties was found in the muscles.     

Strain-induced reorientation of an intramuscular connective tissue network: implications for passive muscle elasticity

The most abundant intramuscular connective tissue component, the perimysium, of bovine M. sternomandibularis muscle was shown to be a crossed-ply arrangement of crimped collagen fibres which reorientate and decrimp on changing muscle fibre sarcomere length. Reorientation of perimysial strands was observed by light microscopy and identification of these strands as collagen fibres was confirmed by high-angle X-ray diffraction. Mean collagen fibre direction with respect to the muscle fibres ranged from approximately 80 degrees at sarcomere length = 1.1 micron to approximately 20 degrees at 3.9 microns. This behaviour was well described by a model of a crimped planar network surrounding a muscle fibre bundle of constant volume but varying length. Modelling of the mechanical properties of the perimysium at different sarcomere lengths produced a load-sarcomere length curve which was in good agreement with the passive elastic properties of the muscle, especially at long sarcomere lengths. It is concluded that the role of the perimysial collagen network is to prevent over-stretching of the muscle fibre bundles.     

Stomatal length and frequency as a measure of ploidy level in black wattle, Acacia mearnsii (de Wild)

The length and frequency of stomata on leaf surfaces were examined as rapid techniques for future identification of ploidy level of Acacia mearnsii (de Wild). Diploid (2 n  = 2 x  = 26) and tetraploid (2 n  = 4 x  = 52) plants were germinated from chipped seed at 25°C and grown under nursery conditions. After one month, measurements showed that the mean stomatal length was 27.17 ± 0.474 µm for diploids and 40.24 ± 0.521 µm for tetraploids and these differed significantly from each other ( P  < 0.001). The frequency of stomata per leaf surface was shown to decrease significantly ( P  < 0.001) as the ploidy level increased, with a mean of 22.11 ± 0.495 for diploids and 10.26 ± 0.495 for tetraploids. It was concluded that stomatal length and stomatal frequency are rapid indirect methods to identify ploidy level in black wattle.  © 2003 The Linnean Society of London, Botanical Journal of the Linnean Society , 2003, 141 , 177–181.     

The diameter and mean sarcomere length of individual muscle fibres.

Studies of the coefficients of variation and the repeatibility of the measurements indicate that a sample of 25 fibres is sufficient to provide an accurate estimate of the mean fibre diameter and sarcomere length of a muscle. There is a significant negative correlation (−0.55) between the diameter and mean sarcomere length of an individual muscle fibre. Because they affect sarcomere length postmortem mechanical influences must be strictly standardised if fibre diameter is to be a reliable parameter of muscle growth.     

The effect of tetraethylammonium and the decrease in the extracellular chloride concentration on membrane depolarization and contraction of skeletal muscle fiber induced by a hyperpotassium solution

1.--The tetraethylammonium (TEA) effects on K+ contracture and membrane depolarization are compared in both crab and frog skeletal muscle fibres. 2.--The mechanical tension of the contracture is reduced by the TEA in frog skeletal muscle fibre; it is increased in crab skeletal fibre. 3.--When no mechanical phenomenon is observed in frog skeletal muscle, the amplitude and the velocity of membrane depolarization induced by an increase of outward K+ concentration is reduced by the TEA. These effects are in opposition in crab muscle fibre. 4.--In crab muscle fibre, the results obtained tend to show that the C1-ions are not distributed on each side of the membrane according to Donnan equilibrium.     

The exine ultrastructure of pollen grains in Gnetum (Gnetaceae) from China and its bearing on the relationship with the ANITA Group

Pollen grains of six species of Gnetum , G. parvifolium , G. hainanense , G. luofuense , G. pendulum , G. cleistostachyum and G. montanum , collected from China were examined using light, scanning and transmission electron microscopy. Pollen grains of Gnetum are subspheroidal or irregular-apolar, inaperturate, 11.21–22.44 µm in long axis and 9.34–20.47 µm in short axis. The exine surface is covered with spinules, 0.50(0.30–0.71) µm long spaced on average 1.12(0.81–1.46) µm apart. The exine is about 0.55 µm thick and comprises ectexine and endexine. The ectexine includes a thin tectum and an infratectal granular layer. The tectum protrudes outwards, forming the spinules. The endexine is composed of discontinuous lamellae, with lacunae between lamellae. The pollen grains of Gnetum are compared with those of Ephedra and Welwitschia , and also those of the ANITA Group of angiosperms, including Amborellaceae, Nymphaeales, Illiciales, Trimeniaceae and Austrobaileyaceae. The exine ultrastructures of Gnetum , Ephedra and Welwitschia are quite similar, consisting of tectum, granular layer and lamellated endexine. The exine ultrastructure of Gnetum is also similar to that of Nymphaea colorata (Nymphaeaceae) in the transitional region between the proximal and distal poles, but differs from that of Amborellaceae, Illicium religiosum (Illiciaceae), Schisandra (Schisandraceae), Trimeniaceae and Austrobaileyaceae. This comparison of exine ultrastructure provides new evidence for consideration of the relationship between Gnetum and the ANITA Group.  © 2004 The Linnean Society of London, Botanical Journal of the Linnean Society , 2004, 146 , 415–425.     

Residual force enhancement and force depression in human single muscle fibres

Residual force depression (rFD) and residual force enhancement (rFE) are intrinsic contractile properties of muscle. rFD is characterized as a decrease in steady-state isometric force following active shortening compared with a purely isometric contraction at the same muscle length and level of activation. By contrast, isometric force is increased following active lengthening compared to a reference isometric contraction at the same muscle length and level of activation; this is termed rFE. To date, there have been no investigations of rFD and rFE in human muscle fibres, therefore the purpose of this study was to determine whether rFD and rFE occur at the single muscle fibre level in humans. rFD and rFE were investigated in maximally activated single muscle fibres biopsied from the vastus lateralis of healthy adults. To induce rFD, fibres were activated and shortened from an average sarcomere length (SL) of 3.2–2.6 μm. Reference isometric contractions were performed at an average SL of 2.6 μm. To induce rFE, fibres were actively lengthened from an average SL of 2.6–3.2 μm and a reference isometric contraction was performed at an average SL of 3.2 μm. Isometric steady-state force was lower following active shortening (p < 0.05), and higher following active lengthening (p < 0.05), as compared to the reference isometric contractions. We demonstrated rFD and rFE in human single fibres which is consistent with previous animal models. The non-responder phenomenon often reported in rFE studies involving voluntary contractions at the whole human level was not observed at the single fibre level.     

Abnormalities in structure and function of limb skeletal muscle fibres of dystrophic mdx mice.

In this study we have shown that the skeletal muscle fibres from adult (older than 26 weeks) mdx mice have gross structural deformities. We have characterized the onset and age dependence of this feature in mdx mice. The three dimensional structure of these deformities has been visualized in isolated fibres and the orientation of these deformities was determined within the muscle by confocal laser scanning microscopy. We have also shown that the occurrence of morphologically abnormal fibres is greater in muscles with longer fibres (extensor digitorum longus (EDL) and soleus, 6-7.3 mm long), than in muscles with shorter fibres (flexor digitorum brevis (FDB), 0.3-0.4 mm long). A population of post-degenerative fibres, with both central and peripheral nuclei coexistent along the length of the fibre, has also been identified in the muscles studied. We showed that a mild protocol of lengthening (eccentric) contractions (the muscle was stretched by 12% during a tetanic contraction) caused a major reduction in the maximal tetanic force subsequently produced by mdx EDL muscle. In contrast, maximal tetanic force production in normal soleus, normal EDL and mdx soleus muscles was not altered by this protocol. We suggest that the deformed fast glycolytic fibres which are found in adult mdx EDL but not in adult mdx soleus muscles are the population of fibres damaged by the lengthening protocol.     

Growth characteristics of skeletal muscle tissue in Oreochromis niloticus larvae fed on a lysine supplemented diet

The effect of lysine amino acid supplementation on the growth characteristics and morphological pattern of skeletal muscle tissue in Nile tilapia Oreochromis niloticus larvae was evaluated. There were four treatments (T) with increasing levels of lysine supplement (T1 = 0·0%; T2 = 1·1%; T3 = 1·7%; T4 = 4·0%) and one treatment with a commercial diet (T5). In all treatments, morphological and histochemical muscle tissue analyses were similar. Two distinct layers were identified: a superficial red layer, more developed in the lateral line region, formed by fibres with intense to moderate NADH‐TR reaction and strong acid‐stable mATPase activity, and a deep white one, most of the muscle mass, formed by fibres with weak NADH‐TR reaction and strong alkali‐stable mATPase activity. There was an intermediate layer between these two layers with fibres exhibiting either weak acid‐stable or acid‐labile mATPase activity. Body mass increase was significantly higher in T5 than in the lysine treatments (T1–T4). There was no difference in number and diameters of muscle fibres between lysine treatments. In T5, muscle fibre diameter and number were higher. The frequency of red fibres with diameters ≤8 μm was higher in the lysine treatments, and with diameters between 16 and 24 μm, was higher in T5. Most white fibre diameters in T5 were significantly larger than 24 μm and in T1–T4 were between 8 and 16 μm. Cell proliferation was higher in the lysine treatments and muscle growth in T5 was mainly by fibre hypertrophy.     

Studies on the nature of the Z-discs in skeletal muscle fibres of sharks, Etmopterus spinax L. and Galeus melastomus Rafinesque-Schmaltz

The ultrastructural details of Z-discs from red, intermediate, and white axial muscle fibres from the sharks Etmopterus spinax and Galeus melastomus are described. Red fibre Z-discs contain the most amorphous matrix material, and are thicker (100–115 nm) than intermediate (85–88 nm) and white Z-discs (75–80 nm). Four sets of oblique bars extend tangentially from each thin filament. In each set two bars are present, those of white fibres are close together (approximately 5 nm), while those of red fibres are separated by a distance of 15 nm. A model of shark Z-disc structure is proposed. The denaturation (heat transition) temperatures of the muscle proteins were studied by differential scanning calorimetry (DSC). The heat transitions of collagen, actin, and myosin were identified; the actin heat transition temperature increased in the sequence red, intermediate, and white. The total protein pattern of red and white muscle were studied by SDS electrophoresis. A protein with a molecular weight of about 55000 may represent a Z-disc protein.     

Behaviour of the crossbridges in stretched or compressed muscle fibres

The maximum chord of the myosin heads is comparable to the closest surface-to-surface spacing between the myofilaments in a muscle at the slack length. Therefore, when the sarcomere length increases or when the fibre is compressed, the surface-to-surface myofilament spacing becomes lower than the head long axis. We conclude that, in stretched or compressed fibres, some crossbridges cannot attach, owing to steric hindrance. When the amount of compression is limited, this hindrance may be overcome by a tilting of the heads in the plane perpendicular to the filament axes; in this case, there is no consequence as concerns the crossbridge properties. In highly compressed fibres, the crossbridges become progressively hindered and all the crossbridges are hindered for an axis-to-axis spacing representing about 60% of the spacing observed under zero external osmotic pressure. In this case, both the isometric tension and the ATPase activity of the fibre are zero. In fibres stretched up to 3.77 microns (sarcomere length corresponding to the disappearance of the overlap between the thick and the thin filaments), the ratio of hindered crossbridges over the functional crossbridges may be estimated at about 55%. In stretched fibres, a noticeable proportion of crossbridges are sterically hindered and the crossbridges performance (e.g. constants of attachment and detachment) depends on filament spacing, i.e. on sarcomere length. Therefore, we think it is probably impossible to consider the crossbridges as independent force converters, since this idea requires that the crossbridge properties are independent of sarcomere length. In this connection, all the experiments performed on osmotically compressed fibres are of major importance for the understanding of the true mechanisms of muscle contraction.     

Expression of titin isoforms in red and white muscle fibres of carp (Cyprinus carpio L.) exposed to different sarcomere strains during swimming

Titin (also known as connectin) is a striated-muscle-specific protein that spans the distance between the Z- and M-lines of the sarcomere. The elastic segment of the titin molecule in the I-band is thought to be responsible for developing passive tension and for maintaining the central position of thick filaments in contracting sarcomeres. Different muscle types express isoforms of titin that differ in their molecular mass. To help to elucidate the relation between the occurrence of titin isoforms and the functional properties of different fibre types, we investigated the presence of different titin isoforms in red and white fibres of the axial muscles of carp. Gel electrophoresis of single fibres revealed that the molecular mass of titin was larger in red than in white fibres. Fibres from anterior and posterior axial muscles were also compared. For both white and red fibres the molecular mass of titin in posterior muscle fibres was larger than in anterior muscle fibres. Thus, the same fibre type can express different titin isoforms depending on its location along the body axis. The contribution of titin to passive tension and stiffness of red anterior and posterior fibres was also determined. Single fibres were skinned and the sarcomere length dependencies of passive tension and passive stiffness were determined. Measurements were made before and after extracting thin and thick filaments using relaxing solutions with 0.6 mol · l⁻¹ KCl and 1 mol · l⁻¹ KI. Tension and stiffness measured before extraction were assumed to result from both titin and intermediate filaments, and tension after extraction from only intermediate filaments. Compared to mammalian skeletal muscle, intermediate filaments developed high levels of tension and stiffness in both posterior and anterior fibres. The passive tension-sarcomere length curve of titin increased more steeply in red anterior fibres than in red posterior fibres and the curve reached a plateau at a shorter sarcomere length. Thus, the smaller titin isoform of anterior fibres results in more passive tension and stiffness for a given sarcomere strain. During continuous swimming, red fibres are exposed to larger changes in sarcomere strain than white fibres, and posterior fibres to larger changes in strain than anterior fibres. We propose that sarcomere strain is one of the functional parameters that modulates the expression of different titin isoforms in axial muscle fibres of carp. Accepted: 7 May 1997     

Seasonal differences in muscle fibre recruitment of pilchard larvae in the north‐western Mediterranean

The recruitment of slow and fast myotomal muscle fibres with respect to growth in body length in European pilchard larvae Sardina pilchardus [(3·5–13·5 mm standard length ( L _S)] was significantly higher in November 1998 than February 1999. This resulted in a significant seasonal difference in the relationship between fibre number and L _S, particularly for the fast muscle. Mean sea surface temperature was c . 6° C higher in November than February, whereas the mean abundance of potential prey items (copepod nauplii) was comparable between cruises. Laboratory and field data obtained from other clupeid species have indicated the importance of early thermal experience on muscle fibre recruitment patterns. Differences in average sea temperature therefore provide a plausible mechanism for the observed seasonal differences in muscle growth characteristics.     

Growth and muscle cellularity of diploid and triploid Atlantic cod (Gadus morhua Linnaeus, 1758) larvae

The aim of this study was to compare somatic growth and muscle fibre development in diploid and triploid siblings of Atlantic cod (Gadus morhua Linnaeus, 1758) during the larval stage. Newly hatched larvae were transferred into 200‐L tanks, three tanks per ploidy group (70 larvae L^?1, continuous light, gradually increasing seawater temperature 7–11°C and flow rates 50–117 L h^?1). Larvae were fed rotifers from 2 to 22 days post hatch (dph), Artemia 19–31 dph and weaned onto a microparticulate diet from 26 dph until the end of the experiment. Measurements of growth (dry weight, standard length) and muscle cellullarity were taken at intervals between 1 and 44 dph. Ploidy groups showed a similar performance throughout the trial, although a marked stagnation in growth was observed for triploids during the weaning from Artemia onto dry feed. Overall, diploid and triploid cod larvae showed a similar development in muscle fibre growth pattern during the experimental period. For both groups, the total number of fast muscle fibres showed a 10‐fold increase (from 384 to 3462), whereas the diameter of fast fibre increased from 8.9 to 13.3 μm (mean number from all treatments). Thus, a temporary but significant effect of triploidy on fast muscle fibre growth pattern was observed in 19 dph larvae in terms of fibre size and number, with triploids showing larger mean fast fibre diameter (11.62 ± 0.63 vs. 10.05 ± 0.34) and a lower number of fibres with a diameter <5 μm than their diploid siblings. Thus, this was found to be related to larvae size and to the differences in total fast fibre cross sectional areas rather than to ploidy status. Overall, our results suggest possible deficiencies in nutrients’ digestion and absorption of triploid cod larvae particularly during the transitional period from live food to inert diets.     

Twitch and tetanic tension during culture of mature Xenopus laevis single muscle fibres

Investigation of the mechanisms of muscle adaptation requires independent control of the regulating factors. The aim of the present study was to develop a serum-free medium to culture mature single muscle fibres of Xenopus laevis. As an example, we used the culture system to study adaptation of twitch and tetanic force characteristics, number of sarcomeres in series and fibre cross-section. Fibres dissected from m. iliofibularis (n = 10) were kept in culture at a fibre mean sarcomere length of 2.3 microm in a culture medium without serum. Twitch and tetanic tension were determined daily. Before and after culture the number of sarcomeres was determined by laser diffraction and fibre cross-sectional area (CSA) was determined by microscopy. For five fibres twitch tension increased during culture and tetanic tension was stable for periods varying from 8 to 14 days ('stable fibres'), after which fibres were removed from culture for analysis. Fibre CSA and the number of sarcomeres in series remained constant during culture. Five other fibres showed a substantial reduction in twitch and tetanic tension within the first five days of culture ('unstable fibres'). After 7-9 days of culture, three of these fibres died. For two of the unstable fibres, after the substantial force reduction, twitch and tetanic tension increased again. Finally at day 14 and 18 of culture, respectively, the tensions attained values higher than their original values. For stable fibres, twitch contraction time, twitch half-relaxation time and tetanus 10%-relaxation time increased during culture. For unstable fibres these parameters fluctuated. For all fibres the stimulus threshold fluctuated during the first two days, and then remained constant, even for the fibres that were cultured for at least two weeks. It is concluded that the present culture system for mature muscle fibres allows long-term studies within a well-defined medium. Unfortunately, initial tetanic and twitch force are poor predictors of the long-term behaviour of the fibres.     

On the absence of circular muscle elements in the body wall of Dysponetus pygmaeus (Chrysopetalidae, 'Polychaeta', Annelida)

The annelid body wall generally comprises an outer layer of circular muscle fibres and an inner layer of longitudinal muscle fibres as well as parapodial and chaetal muscles. An investigation of Dysponetuspygmaeus (Chrysopetalidae) with confocal laser scanning microscopy showed that circular muscles are entirely absent. Further studies indicate that this feature is characteristic for all Chrysopetalidae. A scrutiny of the literature showed a similar situation in many other polychaetes. This lack of circular muscle fibres may either be due to convergence or represent a plesiomorphic character. Since circular muscles are very likely important for burrowing forms but not necessary for animals which proceed by movements of their parapodial appendages or cilia, this problem is also related to the question of whether the ancestral polychaete was epi‐ or endobenthic.