Response of insect muscle to denervation—II. Changes in neuromuscular transmission

The effects of motor-nerve section on neuromuscular transmission in the locust extensor tibiae preparation have been investigated by intracellular recording. Impulse transmission failed between the ninth and twenty-fourth post-operative days, in the region of the nerve-muscle synapse. Before impulse transmission failed, abnormally large depolarizing and hyperpolarizing responses to stimulation of the ‘slow’ axons, and prolonged depolarizing responses to the ‘fast’ axon were recorded from many muscle fibres. At the time of impulse transmission failure the discharge of spontaneous miniature post-synaptic potentials underwent a number of complicated changes leading to the appearance of ‘giant’ miniature potentials up to 10 mV in height. The discharge persisted apparently for a few days after the muscle became neurally inexcitable. When the discharge finally stopped the fibres became permanently electrically silent; no evidence was obtained for a resumption of miniature activity during the later post-operative days. The cessation of the miniature discharge is apparently correlated to the decline of muscle-resting potential observed in denervated insect muscle and the possibility that the transmitter, or a substance released along with it, maintains the resting potential of the muscle at its normal level is discussed.     

Airway smooth muscle electrophysiology in a state of flux?

Activation of chloride currents and release of internally sequestered Ca(2+) in airway smooth muscle have long been associated with excitation and contraction. Surprisingly, however, two recent publications (Deshpande DA, Wang WC, McIlmoyle EL, Robinett KS, Schillinger RM, An SS, Sham JS, Liggett SB. Nat Med 16: 1299-1304, 2010; Gallos G, Yim P, Chang S, Zhang Y, Xu D, Cook JM, Gerthoffer WT, Emala CW Sr. Am J Physiol Lung Cell Mol Physiol 302: L248-L256, 2012) have linked both events to relaxation. This begs a closer look at our understanding of airway smooth muscle electrophysiology and its contribution to excitation-contraction coupling. This Editorial Focus highlights those two aforementioned studies and several other equally paradoxical findings and proposes some possible reinterpretations of the data and/or new directions of research in which the answers might be found.     

Response of insect muscle to denervation—I. Resting potential changes

The effects of motor-nerve section on the resting potential of locust leg muscle have been examined using the intracellular microelectrode technique. Measurements obtained from a large number of preparations indicated that during the first two post-operative days the resting potential of denervated muscle falls below normal. The potential then rises rapidly during the next 2–6 days, only to fall once again to a level about 10–30 per cent below normal. The significance of these findings is discussed in relation to previous investigations.     

Changes of macromolecular organizations in nonjunctional sarcolemmas after cross—innervation—a study of fast—and slow—twitch muscle fiberes in rats

The purpose of the present study was to analyse the changes of macromolecular organizations in nonjunctional sarcolemmas of different types of skeletal muscle fibres after cross-innervation.In normal rate the mean density of square arrays(6nm particles organized in orthogonal of square arrays(6nm particles organized in othogonal arrays)was 9.02/μm^2 for the nonjunctional sarcolemmas of fast-twitch extensor digitorum longus(control EDL,CE)muscle fibres and 0.34/μm^2 for the nunjunctional sarcolemmas of slow-twitch soleus(control SOL,CS)muscle fibres.After cross-innervation between the fast-twitch EDL and slow-twitch SOL muscle fibres by slow and fast muscle nerves respectively for three months ,the mean density was 0.45/μm^2 for the nonjunctional sarcolemmas of the operated EDL(OE)and 8.3/μm^2 for the nonjunctional sarcolemmas of the operated SOL(OS).This indicates that the cross-innervation causes a reciprocal transformation of the number and distribution of such macromolecular organizations in the electrically excitable nonjunctional sarcolemmas.     

Role of protein kinase C in oxidant — mediated activation of phospholipase A2 in rabbit pulmonary arterial smooth muscle cells

The present study was undertaken to test the hypothesis that activation of cell membrane associated protein kinase C (PKC) plays a role in stimulating cell membrane associated phospholipase A₂ (PLA₂) activity, and subsequent liberation of arachidonic acid (AA) under exposure of rabbit pulmonary arterial smooth muscle cells to the oxidant hydrogen peroxide (H₂O₂). Exposure of the smooth muscle cells to H₂O₂ dose-dependently stimulates [¹⁴C] AA release, and enhances the cell membrane associated PLA₂ activity. Pretreatment of the cells with protein kinase C (PKC) inhibitors H₇ and sphingosine prevent the cell membrane associated PLA₂ activity, and AA release caused by H₂O₂. Treatment of the smooth muscle cells with H₂O₂ stimulates the cell membrane associated PKC activity. Pretreatment of the cells with an antioxidant vitamin E prevents H₂O₂ caused stimulation of the cell membrane associated PKC activity. The cell membrane associated PLA₂ and PKC activities correlate linearly. These results suggest that H₂O₂ caused stimulation of the smooth muscle cell membrane associated PLA₂ activity, and subsequent liberation of AA can occur through an increase in the activity of the cell membrane associated PKC. (Mol Cell Biochem122: 9–15, 1993)Abbreviations AA Arachidonic Acid - PLA₂ Phospholipase A₂ - PKC Protein Kinase C - PBS Phosphate Buffered Saline - HBPS Hank's Buffered Physiological Saline - HEPES 4-(2-Hydroxyethyl)-1-Piperazine N-2-Ethanesulfonate - FCS Fetal Calf Serum - ATP Adenosine Triphosphate - H₇ 1-(5-isoquinolinesulfonyl)-2-methyl-piperazine - DMEM Dulbecco's Modified Eagles Medium - TCA Trichloroacetic Acid     

A change of heart: oxidative stress in governing muscle function?

Biophysical Reviews - Redox/cysteine modification of proteins that regulate calcium cycling can affect contraction in striated muscles. Understanding the nature of these modifications would present...     

Redundancy or heterogeneity in the electric activity of the biceps brachii muscle? Added value of PCA-processed multi-channel EMG muscle activation estimates in a parallel-fibered muscle

Conventional bipolar EMG provides imprecise muscle activation estimates due to possibly heterogeneous activity within muscles and due to improper alignment of the electrodes with the muscle fibers. Principal component analysis (PCA), applied on multi-channel monopolar EMG yielded substantial improvements in muscle activation estimates in pennate muscles. We investigated the degree of heterogeneity in muscle activity and the contribution of PCA to muscle activation estimates in biceps brachii (BB), which has a relatively simply parallel-fibered architecture. EMG-based muscle activation estimates were assessed by comparison to elbow flexion forces in isometric, two-state isotonic contractions in eleven healthy male subjects. Monopolar EMG was collected over the entire surface of the BB with about 63 electrodes. Estimation quality of different combinations of EMG channels showed that heterogeneous activation was found mainly in medio-lateral direction, whereas adding channels in the longitudinal direction added largely redundant information. Multi-channel bipolar EMG amplitude improved muscle activation estimates by 5–14% as compared to a single bipolar. PCA-processed monopolar EMG amplitude yielded a further improvement of (12–22%). Thus multi-channel EMG, processed with PCA, substantially improves the quality of muscle activation estimates compared conventional bipolar EMG in BB.     

Is maximum stimulation intensity required in the assessment of muscle activation capacity?

Voluntary activation assessment using the interpolation twitch technique (ITT) has almost invariably been done using maximal stimulation intensity, i.e., an intensity beyond which no additional joint moment or external force is produced by increasing further the intensity of stimulation. The aim of the study was to identify the minimum stimulation intensity at which percutaneous ITT yields valid results. Maximal stimulation intensity and the force produced at that intensity were identified for the quadriceps muscle using percutaneous electrodes in eight active men. The stimulation intensities producing 10–90% (in 10% increments) of that force were determined and subsequently applied during isometric contractions at 90% of maximum voluntary contraction (MVC) via twitch doublets. Muscle activation was calculated with the ITT and pain scores were obtained for each stimulation intensity and compared to the respective values at maximum stimulation intensity. Muscle activation at maximal stimulation intensity was 91.6 (2.5)%. The lowest stimulation intensity yielding comparable muscle activation results to maximal stimulation was 50% (88.8 (3.9)%, p < 0.05). Pain score at maximal stimulation intensity was 6.6 (1.5) cm and it was significantly reduced at 60% stimulation intensity (3.7 (1.5) cm, p < 0.05) compared to maximal stimulation intensity. Submaximal stimulation can produce valid ITT results while reducing the discomfort obtained by the subjects, widening the assessment of ITT to situations where discomfort may otherwise impede maximal electrostimulation.     

Re-emergence of glanders in India — Report of Maharashtra state

Glanders, a notifiable highly contagious disease primarily of equids, is a disease of high zoonotic importance. Caused by gram-negative bacillus, Burkholderia mallei, the disease was restricted to certain pockets of India with sporadic cases. Recently, a major outbreak of glanders occurred in India starting from Maharashtra. Following clinical signs and symptoms and laboratory investigations on serum, nasal swab and pus swab samples, it was confirmed as glanders among equines in Pune and Panchgani areas of Maharashtra. One pus sample and three nasal swabs yielded B. mallei isolates while 23 serum samples were found positive for glanders by complement fixation test (CFT). The disease was successfully controlled in the state by following strategies for prevention of spread of the disease to other areas in accordance with Glanders and Farcy Act, 1899. Follow up of the occurrence in Maharashtra revealed negative status based on testing and physical surveillance on more than 3,500 equines thereafter. Investigations indicated that the nidus of infection may be present elsewhere in North India.     

Studies on the degeneration of the flight muscles of alate aphids—II Histology and control of muscle breakdown

Within a few days of settling down on a host plant, alate aphids of most species undergo a change in metabolism which becomes manifest in breakdown of the flight muscles, resumption of embryo development in the ovarioles, and hypertrophy of the fat body. In this paper an account is given of the histology of flight muscle breakdown, and experimental evidence is presented which suggests that the change in metabolism in the aphid is brought about by the secretion of a hormone. Muscle breakdown and fat body hypertrophy could be initiated in aphids in response to abnormal afferent stimuli, as when aphids were tied down on inert surfaces.     

Role of ubiquitin‐proteasome‐dependent proteolytic process in degradation of muscle protein from diabetic rabbits

The activity of ATP, ubiquitin (Ub)dependent proteases partially purified from skeletal muscle (psoas) from alloxan diabetic rabbits was determined at different periods of insulin deficiency. Two days after alloxan injection, no change was observed in the activity of ATP, Ubdependent proteases, but this activity increased 3 and 5 days after diabetes induction, attaining 181% of control values on the 5th day. However, after this early rise, the activity of muscle ATP, Ubdependent proteases decreased, returning to values that did not differ significantly from controls 7 and 10 days after alloxan injection. After 15 days, the activity of these proteases was 57% lower than in muscle from control rabbits. Both the initial increase and the subsequent fall in the activity of the enzymes were prevented by insulin treatment of alloxan diabetic rabbits. The data suggest that Ubproteasomedependent proteolysis have an important role in the control of muscle protein degradation and may be regulated by insulin.     

Effects of training and weight support on muscle activation in Parkinson’s disease

The aim of this study was to investigate the effect of high-intensity locomotor training on knee extensor and flexor muscle activation and adaptability to increased body-weight (BW) support during walking in patients with Parkinson’s disease (PD). Thirteen male patients with idiopathic PD and eight healthy participants were included. The PD patients completed an 8-week training program on a lower-body, positive-pressure treadmill. Knee extensor and flexor muscles activation during steady treadmill walking (3 km/h) were measured before, at the mid-point, and after training. Increasing BW support decreased knee extensor muscle activation (normalization) and increased knee flexor muscle activation (abnormal) in PD patients when compared to healthy participants. Training improved flexor peak muscle activation adaptability to increased (BW) support during walking in PD patients. During walking without BW support shorter knee extensor muscle off-activation time and increased relative peak muscle activation was observed in PD patients and did not improve with 8 weeks of training. In conclusion, patients with PD walked with excessive activation of the knee extensor and flexor muscles when compared to healthy participants. Specialized locomotor training may facilitate adaptive processes related to motor control of walking in PD patients.     

Decay kinetics and quantum yields of fluorescence in photosystem I from Synechococcus elongatus with P700 in the reduced and oxidized state: are the kinetics of excited state decay trap-limited or transfer-limited?

Transfer and trapping of excitation energy in photosystem I (PS I) trimers isolated from Synechococcus elongatus have been studied by an approach combining fluorescence induction experiments with picosecond time-resolved fluorescence measurements, both at room temperature (RT) and at low temperature (5 K). Special attention was paid to the influence of the oxidation state of the primary electron donor P700. A fluorescence induction effect has been observed, showing a approximately 12% increase in fluorescence quantum yield upon P700 oxidation at RT, whereas at temperatures below 160 K oxidation of P700 leads to a decrease in fluorescence quantum yield ( approximately 50% at 5 K). The fluorescence quantum yield for open PS I (with P700 reduced) at 5 K is increased by approximately 20-fold and that for closed PS I (with P700 oxidized) is increased by approximately 10-fold, as compared to RT. Picosecond fluorescence decay kinetics at RT reveal a difference in lifetime of the main decay component: 34 +/- 1 ps for open PS I and 37 +/- 1 ps for closed PS I. At 5 K the fluorescence yield is mainly associated with long-lived components (lifetimes of 401 ps and 1.5 ns in closed PS I and of 377 ps, 1.3 ns, and 4.1 ns in samples containing approximately 50% open and 50% closed PS I). The spectra associated with energy transfer and the steady-state emission spectra suggest that the excitation energy is not completely thermally equilibrated over the core-antenna-RC complex before being trapped. Structure-based modeling indicates that the so-called red antenna pigments (A708 and A720, i.e., those with absorption maxima at 708 nm and 720 nm, respectively) play a decisive role in the observed fluorescence kinetics. The A720 are preferentially located at the periphery of the PS I core-antenna-RC complex; the A708 must essentially connect the A720 to the reaction center. The excited-state decay kinetics turn out to be neither purely trap limited nor purely transfer (to the trap) limited, but seem to be rather balanced.     

High K(m) of oxidative phosphorylation for ADP in skinned muscle fibers: where does it stem from?

Mitochondria insaponin-skinned cardiac fiber bundles were reported to have an order ofmagnitude lower apparent affinity to ADP than isolatedmitochondria. Although ADP was measured outside the bundles, it wasthought that the low affinity was not caused by diffusion gradientsbecause of relatively short diffusion distances. Here we test thehypothesis that considerable ADP diffusion gradients exist and can bediminished by increasing the intrafiber ADP production rate. Weincreased the ADP-producing activity in rat heart skinned fiber bundlesby incubating with 100 IU/ml yeast hexokinase and glucose.Consequently, we observed a significant decrease of the apparentMichaelis constant (K_m) to ADP of therespiration rate of bundles from 216 ± 59 to 50 ± 9 µM.Fitting the results with a mathematical model, we estimated theK_m of mitochondria in the bundles to be 25 µM.We conclude that the affinity to ADP of in situ mitochondria in heartis of the same order of magnitude as that of isolated mitochondria.

     

Viscoelastic properties of passive skeletal muscle in compression—Cyclic behaviour

Skeletal muscle relaxation behaviour in compression has been previously reported, but the anisotropic behaviour at higher loading rates remains poorly understood. In this paper, uniaxial unconfined cyclic compression tests were performed on fresh porcine muscle samples at various fibre orientations to determine muscle viscoelastic behaviour. Mean compression level of 25% was applied and cycles of 2% and 10% amplitude were performed at 0.2–80 Hz. Under cycles of low frequency and amplitude, linear viscoelastic cyclic relaxation was observed. Fibre/cross-fibre results were qualitatively similar, but the cross-fibre direction was stiffer (ratio of 1.2). In higher amplitude tests nonlinear viscoelastic behaviour with a frequency dependent increase in the stress cycles amplitude was found (factor of 4.1 from 0.2 to 80 Hz).The predictive capability of an anisotropic quasi-linear viscoelastic model previously fitted to stress-relaxation data from similar tissue samples was investigated. Good qualitative results were obtained for low amplitude cycles but differences were observed in the stress cycle amplitudes (errors of 7.5% and 31.8%, respectively, in the fibre/cross-fibre directions). At higher amplitudes significant qualitative and quantitative differences were evident. A nonlinear model formulation was therefore developed which provided a good fit and predictions to high amplitude low frequency cyclic tests performed in the fibre/cross-fibre directions. However, this model gave a poorer fit to high frequency cyclic tests and to relaxation tests. Neither model adequately predicts the stiffness increase observed at frequencies above 5 Hz.Together with data previously presented, the experimental data presented here provide a unique dataset for validation of future constitutive models for skeletal muscle in compression.     

Growth,starvation and enzyme activity in white muscle of atlantic cod: at what point do muscle metabolic capacities change?

Muscle protein decreases only during prolonged starvation of Atlantic cod (Gadus morhua, Gadidae), but in the absence of protein renewal, muscle metabolic capacities may decrease before marked loss of muscle protein. This study aimed at elucidating the threshold at which decreases in growth and condition reduce muscle metabolic capacities, as well as identifying the indicators that best explain changes in metabolic capacities. To generate a wide spectrum of individual growth rates, condition factors and proximate compositions, cod showing different initial condition were fed or starved for different periods of time. The relationships between muscle proteins and metabolic enzyme activities (LDH and CCO) on one hand, and growth rate, condition factor, hepato- and gonadosomatic index and muscle and liver water and energy contents, on the other hand, were examined through linear regression models. Multiple linear regressions explained a large proportion of the observed variability in proteins and enzyme activities. Changes in LDH and CCO activities were not driven by changes in growth rate. Muscle water was the only significant correlate for both enzymes. Enzyme activities decreased as soon as muscle water began to rise. Increases in water content from 79 to 92% resulted in a 10-fold decrease in LDH and CCO activities.     

An ethanol extract of Artemisia iwayomogi activates PPARδ leading to activation of fatty acid oxidation in skeletal muscle

Although Artemisia iwayomogi (AI) has been shown to improve the lipid metabolism, its mode of action is poorly understood. In this study, a 95% ethanol extract of AI (95EEAI) was identified as a potent ligand of peroxisome proliferator-activated receptorδ (PPARδ) using ligand binding analysis and cell-based reporter assay. In cultured primary human skeletal muscle cells, treatment of 95EEAI increased expression of two important PPARδ-regulated genes, carnitine palmitoyl-transferase-1 (CPT1) and pyruvate dehydrogenase kinase isozyme 4 (PDK4), and several genes acting in lipid efflux and energy expenditure. Furthermore, 95EEAI stimulated fatty acid oxidation in a PPARδ-dependent manner. High-fat diet-induced obese mice model further indicated that administration of 95EEAI attenuated diet-induced obesity through the activation of fatty acid oxidation in skeletal muscle. These results suggest that a 95% ethanol extract of AI may have a role as a new functional food material for the prevention and/or treatment of hyperlipidermia and obesity.     

Interpretation of nonlinear steady state enzyme kinetics—Cyclic and mathematical properties of cooperative,second-site and random pathway models

Nonlinear steady state kinetic patterns are frequently encountered in enzyme studies. Consequently, there is a need to develop procedures for systematically interpreting such data. This paper contributes to this development by identifying a common feature in nonlinear systems and by showing that quite different models commonly in use give very similar mathematical functions.Identical or similar cycles can result from quite different chemical events in enzyme mechanisms, cooperativity, second sites and random pathways. Such cycles can account for many of the observed nonlinear patterns, i.e., power functions, substrate activation and inhibition. Therefore nonlinear steady state kinetics generally requires the presence of a cycle(s) in the mechanism without specifying the underlying chemical events giving rise to that cycle(s).Rate equations for cooperative, second-site and random pathway models are derived and shown to yield virtually identical mathematical functions. Thus empirical equations composed of these functions can be used to represent nonlinear kinetic data without specifying the underlying chemical events.     

Growth, starvation and enzyme activity in white muscle of atlantic cod: at what point do muscle metabolic capacities change?

Muscle protein decreases only during prolonged starvation of Atlantic cod (Gadus morhua, Gadidae), but in the absence of protein renewal, muscle metabolic capacities may decrease before marked loss of muscle protein. This study aimed at elucidating the threshold at which decreases in growth and condition reduce muscle metabolic capacities, as well as identifying the indicators that best explain changes in metabolic capacities. To generate a wide spectrum of individual growth rates, condition factors and proximate compositions, cod showing different initial condition were fed or starved for different periods of time. The relationships between muscle proteins and metabolic enzyme activities (LDH and CCO) on one hand, and growth rate, condition factor, hepato- and gonadosomatic index and muscle and liver water and energy contents, on the other hand, were examined through linear regression models. Multiple linear regressions explained a large proportion of the observed variability in proteins and enzyme activities. Changes in LDH and CCO activities were not driven by changes in growth rate. Muscle water was the only significant correlate for both enzymes. Enzyme activities decreased as soon as muscle water began to rise. Increases in water content from 79 to 92% resulted in a 10-fold decrease in LDH and CCO activities.