Cryo-EM structures of cardiac thin filaments reveal the 3D architecture of troponin

Troponin is an essential component of striated muscle and it regulates the sliding of actomyosin system in a calcium-dependent manner. Despite its importance, the structure of troponin has been elusive due to its high structural heterogeneity. In this study, we analyzed the 3D structures of murine cardiac thin filaments using a cryo-electron microscope equipped with a Volta phase plate (VPP). Contrast enhancement by a VPP enabled us to reconstruct the entire repeat of the thin filament. We determined the orientation of troponin relative to F-actin and tropomyosin, and characterized the interactions between troponin and tropomyosin. This study provides a structural basis for understanding the molecular mechanism of actomyosin system.     

Accumulation of a tumor suppressor p53 protein in rat muscle during a space flight

We previously reported that the space environment consisting of microgravity and space radiation induced an increased level of p53 protein, a tumor suppressor gene product, in rat skin. Here, we report the increase of p53 protein in the muscles of rats that traveled into space. Rats were divided into three groups. The first group remained on earth (VC), and did not show any change in p53 protein level. The second group made a 14-day flight into space on the Second Spacelab Life Science (SLS-2) Mission (F). The third group was experimentally subjected to the same kinds of stress as those in the second group without making a space flight (SC). F and SC rats were sacrificed on day zero (F-0, SC-0) and day nine (F-9, SC-9) after return from space. F-0 rats showed a 1.5-fold increase in p53 protein level compared with that of SC-0 rats, whereas, F-9 rats showed a 1.35-fold increase in p53 protein compared with that of SC-9 rats. These results suggest that the accumulation of cellular p53 protein induced by space environments occurs not only in rat skin cells, but also in rat muscle cells.     

A comparison of electrical activity in the triceps surae at maximum isometric contraction with the knee and ankle at various angles

The purpose of this study was to test the endurance of the soleus muscle, and to examine the joint position at which it is most active, while simultaneously suppressing the activity of the gastrocnemius. Ten young males performed maximum isometric contraction of the triceps surae for 100 s, and the endurance and plantar flexion torque of this muscle were measured at various angles of the knee and ankle joints. The electromyogram was measured simultaneously and subsequently converted into integrated electromyogram (IEMG) values. With the knee flexed at 130 degrees, the rate of change in IEMG values for the soleus (0.454% x s(-1)) with the ankle in a neutral position was significantly higher than that for the medial and lateral gastrocnemius. Both with the ankle dorsiflexed at 10 degrees and in the neutral position, the rate of change in IEMG for the soleus was significantly higher with the knee flexed at 90 degrees and 130 degrees than with the knee fully extended. With the knee flexed at 90 degrees and 130 degrees, the IEMG activity of the soleus during the initial (5-10 s) and final 5 s tended to be higher than those for the medial and lateral gastrocnemius, regardless of the ankle joint position. We conclude that the position in which the soleus acts most selectively during a sustained maximum isometric contraction of the triceps surae is with the ankle in a neutral position and the knee flexed at 130 degrees.     

Enzyme activity of rat tibialis anterior muscle differs between treatment with triamcinolone and prednisolone and nutritional deprivation

The maximal activity of a selection of enzymes involved in muscle carbohydrate handling, citric acid cycle and fatty acyl beta-oxidation were studied after treatment with the fluorinated corticosteroid triamcinolone and compared to a similar treatment of the non-fluorinated corticosteroid prednisolone in an equipotent anti-inflammatory dose. Furthermore, because triamcinolone causes loss of body mass and muscle wasting, the effects of triamcinolone were investigated relative to a control group, with the same loss of body mass, due to nutritional deprivation. The study was performed in male Wistar rats in the following treatment groups: TR, triamcinolone treatment (0.25 mg x kg(-1) x day(-1) for 2 weeks), which resulted in a reduction of body mass (24%); ND, nutritional deprivation (30% of normal daily food intake for 2 weeks) resulting in a similar (24%) decrease of body mass as TR; PR, prednisolone treatment (0.31 mg x kg(-1) x day(-1) for 2 weeks), with a 10% increase in body mass; FF, free-fed control group, with a 12% increase in body mass in 2 weeks. Compared to FF, TR induced an increase in phosphofructokinase (PFK) activity (P < 0.01), glycogen synthase [GS(i + d)] activity (P < 0.05) and glycogen content (P < 0.01) in the tibialis anterior muscle. The PR and ND caused no alterations in PFK or citrate synthase (CS) activity compared to FF. Compared to PR, TR induced an increase in PFK (P < 0.01), CS (P < 0.05) and GS(i + d) activity (P < 0.01). Both TR and PR caused an increased muscle glycogen content, being more pronounced in TR (P < 0.05). Compared to ND, TR induced an increased CS (P < 0.05) and GS(i + d) activity (P < 0.01) and glycogen content (P < 0.01). The ND resulted in a decreased glycogen content compared to FF (P < 0.05). None of the treatments affected the activity of glycogen phosphorylase, beta-hydroxyacyl coenzyme A dehydrogenase and lactate dehydrogenase. It was concluded that corticosteroids led to an increased muscle glycogen content; however, the changes in the enzymes of carbohydrate metabolism were corticosteroid type specific and did not relate to undernutrition, which accompanied the triamcinolone treatment.     

Effect of exhausting stretch-shortening cycle exercise on the time course of mechanical behaviour in the drop jump: possible role of muscle damage

The purpose of the present study was to investigate the effect of stretch-shortening-cycle-induced muscle damage on the time course of mechanical behaviour in the drop jump. Ten healthy male subjects performed submaximal stretch-shortening cycle (SSC) exercise on a special sledge apparatus. Exhaustion occurred on average within 3 min. A drop jump (DJ) test from a 50-cm height was performed before and immediately after the sledge exercise as well as 2 h, 2 days and 4 days later. The fatigue exercise showed relatively high blood lactate concentration [12.5 (SD 2.6) mmol x l(-1)] and an increase of serum creatine kinase (CK) activity delayed by 2 days [540 (SD 407) U x l(-1)]. The initial decline in the jump performance (before - immediately after) was related negatively to the early recovery in performance (immediately after 2 h) (P < 0.05). The early recovery of the knee joint moment at the end of stretch showed a negative correlation to the delayed decrease in DJ performance (2 h 2 days) (P < 0.01). Thus, the DJ performance showed an initial decline followed by an early recovery and a secondary decline. Both the initial decline and early recovery in the knee joint moment at the end of stretch were related to the corresponding initial (after 2 h) (P < 0.05) and secondary increases (2 h - 2 days) (P < 0.01) in CK. It is suggested that the early recovery as well as the initial decline in the knee joint function could depend on the degree of muscle damage. Delayed decrease in initial stiffness (2 h - 2 days) was negatively related to the corresponding changes in the knee joint angle at touch down in DJ (P < 0.001). These interactions would imply that the decrease in the stiffness regulation and the modulation of the prelanding motor control might be attributable to secondary muscle damage during 2 days after the SSC exercise. Therefore, it may be suggested that the changes in the DJ performance after the exhausting SSC exercise accompany the progress of muscle damage observed by the corresponding increase in serum CK concentration and the corresponding deterioration of stiffness regulation and motor control in DJ.     

Physiological factors in stretcher carriage performance

In this study we examined the anthropometric and physiological factors that may account for the ability to carry a casualty on a stretcher. Eleven young soldiers were pretested to obtain their anthropometry, body composition, physical fitness, and muscle cross-sectional areas. They then performed a two-person manual carry of a stretcher containing an 82-kg manikin while walking on a treadmill at a speed of 4.8 km/h. Subjects walked until volitional fatigue, as indicated by slippage of the stretcher from their hands. Average (SD) carriage time was 2.7(1.4) min with a range of 1.4-6.4 min. A stepwise multiple linear regression revealed that forearm bone-plus-muscle cross-sectional area, thigh muscle cross-sectional area, and push-up performance accounted for most of the variance in hand carriage time (r2 = 0.99, P < 0.001). These data suggest that muscle cross-sectional area and upper-body muscular endurance are important physiological factors in the ability to carry a loaded stretcher by hand.     

Determination of myoglobin concentration and oxidative capacity in cryostat sections of human and rat skeletal muscle fibres and rat cardiomyocytes

Myoglobin plays various roles in oxygen supply to muscle mitochondria. It is difficult, and in some cases impossible, to study the relationship between the myoglobin concentration and the oxidative capacity of individual muscle cells because myoglobin has to be fixed in situ whereas determination of oxidative capacity, for example, succinate dehydrogenase activity, requires unfixed cryostat sections. We have investigated whether a vapour-fixation technique allows the use of serial sections to study the relationship between myoglobin and succinate dehydrogenase activity. The technique is used to study a rat soleus muscle, two human skeletal muscle biopsies and biopsies of two patients with chronic heart failure, and in a control and hypertrophied rat heart. Staining intensities were quantified by microdensitometry. The absorbance values were calibrated using sections cut from gelatine blocks containing known amounts of myoglobin. The results show that it is possible to use serial sections for the determination of the myoglobin concentration and succinate dehydrogenase activity, and indicate that myoglobin can lead to a substantial reduction (18–60%) of the extracellular oxygen tension required to prevent an anoxic core in muscle cells.     

Evidence for myofibril remodeling as opposed to myofibril damage in human muscles with DOMS: an ultrastructural and immunoelectron microscopic study

The myofibrillar and cytoskeletal alterations observed in delayed onset muscle soreness (DOMS) caused by eccentric exercise are generally considered to represent damage. By contrast our recent immunohistochemical studies suggested that the alterations reflect myofibrillar remodeling (Yu and Thornell 2002; Yu et al. 2003). In the present study the same human muscle biopsies were further analyzed with transmission electron microscopy and immunoelectron microscopy. We show that the ultrastructural hallmarks of DOMS, Z-disc streaming, Z-disc smearing, and Z-disc disruption were present in the biopsies and were significantly more frequent in biopsies taken 2–3 days and 7–8 days after exercise than in those from controls and 1 h after exercise. Four main types of changes were observed: amorphous widened Z-discs, amorphous sarcomeres, double Z-discs, and supernumerary sarcomeres. We confirm by immunoelectron microscopy that the main Z-disc protein alpha-actinin is not present in Z-disc alterations or in the links of electron-dense material between Z-discs in longitudinal register. These alterations were related to an increase of F-actin and desmin, where F-actin was present within the strands of amorphous material. Desmin, on the other hand, was seen in less dense regions of the alterations. Our results strongly support that the myofibrillar and cytoskeletal alterations, considered to be the hallmarks of DOMS, reflect an adaptive remodeling of the myofibrils.     

The activity of abdominal stretch receptors during non-giant swimming in the crayfish<Emphasis Type="Italic"> Cherax destructor</Emphasis> and their role in hydrodynamic efficiency

Recordings were made from the nerve innervating the stretch receptors of the abdominal muscle receptor organs and slow extensor muscles of tethered crayfish, Cherax destructor, during so-called non-giant swimming. The stretch receptors were active during the flexor phase of swimming but the duration and pattern of activity varied from cycle to cycle. Their pattern of firing was modified by the activity of the large accessory neurons which make direct inhibitory synapses upon them. Neither the stretch receptors nor the accessory neurons were active during the extensor phase of the cycle. The timing and extent of tailfan movements during the period of stretch receptor activity were measured from video records before and after the stretch receptor nerves were cut in the second to fifth segments. The promotion of the tailfan during flexion was significantly delayed and the minimum angle to which the uropods were remoted at the end of flexion significantly larger in denervated animals. We propose that afferent information from the stretch receptors coordinates the timing and extent of tailfan movements according to variations in the positioning and movement of the abdominal segments such that the hydrodynamic efficiency of the tailfan is enhanced on a cycle by cycle basis during non-giant swimming.Abbreviations A# abdominal segment number - Acc accessory neuron - LUU large unidentified unit - MRO muscle receptor organ - NGS non-giant swimming - SEMN slow extensor motor neuron - SR stretch receptor neuron