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PubMed1994

Calmodulin and the regulation of smooth muscle contraction.

Calmodulin, the ubiquitous and multifunctional Ca(2+)-binding protein, mediates many of the regulatory effects of Ca2+, including the contractile state of smooth muscle. The principal function of calmodulin in smooth muscle is to activate crossbridge cycling and the development of force in response to a [Ca2+]i transient via the activation of myosin light-chain kinase and phosphorylation of myosin. A distinct calmodulin-dependent kinase, Ca2+/calmodulin-dependent protein kinase II, has been implicated in modulation of smooth-muscle contraction. This kinase phosphorylates myosin light-chain kinase, resulting in an increase in the calmodulin concentration required for half-maximal activation of myosin light-chain kinase, and may account for desensitization of the contractile response to Ca2+. In addition, the thin filament-associated proteins, caldesmon and calponin, which inhibit the actin-activated MgATPase activity of smooth-muscle myosin (the cross-bridge cycling rate), appear to be regulated by calmodulin, either by the direct binding of Ca2+/calmodulin or indirectly by phosphorylation catalysed by Ca2+/calmodulin-dependent protein kinase II. Another level at which calmodulin can regulate smooth-muscle contraction involves proteins which control the movement of Ca2+ across the sarcolemmal and sarcoplasmic reticulum membranes and which are regulated by Ca2+/calmodulin, e.g. the sarcolemmal Ca2+ pump and the ryanodine receptor/Ca2+ release channel, and other proteins which indirectly regulate [Ca2+]i via cyclic nucleotide synthesis and breakdown, e.g. NO synthase and cyclic nucleotide phosphodiesterase. The interplay of such regulatory mechanisms provides the flexibility and adaptability required for the normal functioning of smooth-muscle tissues.

Amino Acid Sequence↗

PubMed1985

Effects of muscle contraction on pulsatile pressure-flow relations in femoral bed.

Femoral arterial pressure-flow relations and vascular impedance were studied during isometric contraction of the gastrocnemius-plantaris muscle group in anesthetized dogs. Contractions were synchronized with the electrocardiogram to occur in the first or second half of the cardiac cycle and included twitches as well as low-, intermediate-, and high-frequency tetanuses. The effects of fatigue and recovery were also documented. Marked changes in pressure and flow waveforms and corresponding femoral arterial input impedance spectra were seen for all contraction modes. Impedance moduli and estimated characteristic impedance were elevated regardless of contraction mode and were associated with fluctuations in impedance phase. All tetanuses placed in the first half of the cardiac cycle produced a striking and consistent reversal of impedance phase for the fundamental harmonic from negative to positive values which decreased with progressive fatigue. During recovery, impedance spectra were unchanged from control spectra. We have demonstrated marked alterations in pressure and flow waveforms and impedance spectral patterns during isometric contraction in the canine hindlimb. These changes may be explained by 1) markedly increased wave reflection as a result of muscle contraction and/or 2) the generation of a retrograde pulse by contracting muscle that fuses with the antegrade pulse of cardiac origin.

Animals↗

PubMed1986

Effects of cholinergic drugs on longitudinal muscle contractions of Fasciola hepatica.

Acetylcholine, cholinergic agonists and acetylcholinesterase inhibitors significantly decrease the amplitude and frequency of spontaneous longitudinal muscle contractions in Fasciola hepatica. In order of their effects on the inhibition of muscle contractions, the cholinergic agonists can be ranked as nicotine greater than carbachol greater than acetylcholine. High calcium ion concentration also causes a significant inhibition of contractions. Atropine, a cholinergic antagonist that acts on muscarinic receptors, significantly increases the amplitude and frequency of spontaneous contractions and completely reverses the effects of cholinomimetic drugs, including nicotine. In adult F. hepatica, the levels of acetylcholine and its precursor choline are 3.14 +/- 0.55 and 13.75 +/- 3.72 pmol/mg wet weight, respectively. The activities of choline acetyltransferase, specific acetylcholinesterase and the nonspecific cholinesterase are 1.25 +/- 0.19, 238.0 +/- 13.0, and 83.0 +/- 33.0 nmol/hr/mg protein, respectively.

Acetylcholine↗

PubMed1991

A new concept for the mechanism of Ca+(+)-regulation of muscle contraction. Implications for physiological and pharmacological approaches to modulate contractile function of myocardium.

Recent development of an experimental protocol to determine kinetics of active cross-bridge turnover is muscle allows analysis of possible Ca+(+)-effects on cross-bridge turnover kinetics. This analysis enabled us to distinguish the two main hypotheses about the mechanism of regulation of muscle contraction. In the first hypothesis, the number of actively turning over cross-bridges is changed, while cross-bridge turnover kinetics are unaffected by Ca++ (regulation by "cross-bridge recruitment"). In the other hypotheses, cross-bridge turnover kinetics are controlled by Ca++, while the number of actively turning over cross-bridges is essentially unaffected (regulation by "rate modulation"). It is found that the major mechanism of regulation of muscle contraction is by a change in the rate constant (fapp) that determines the transition of a cross-bridge from the weak-binding (non-force generating) configuration to its strong-binding (force generating) configuration. It is demonstrated that the concept of "rate modulation" requires reinterpretation of force-pCa relations and of the mechanisms of physiological and pharmacological modulation of force-pCa relations. On this basis, an additional mechanism for positive inotropic interventions is demonstrated which may have advantages over the previously established mechanisms.

Animals↗

PubMed1984

A hypothesis for neural control of the speed of muscle contraction in the mammal.

A hypothesis for neural control of the contractile properties of muscle is presented. The hypothesis is based on two assumptions: i) the motoneurons innervating both the fast- and slow-twitch muscles in the mammal possess an identical neurotrophic substance which accelerates the speed of muscle contraction, and ii) slow contractions of the slow-twitch muscle are induced by the low frequency discharges of the innervating motoneurons. The presence of such a trophic substance in the motoneurons is supported by the prolongation of contraction time in both the fast- and slow-twitch muscles following denervation. Furthermore, a given slow-twitch muscle dually reinnervated by the fast and slow muscle nerves contracts at the same speed regardless of which nerve is stimulated, if identical neural activity patterns are chronically imposed upon the two nerves. The presence of effective neural activity patterns in the motoneurons innervating the slow-twitch muscle is supported by an increase in the contraction speed of the slow-twitch muscle following virtual elimination of motoneuron activity by transection of the spinal cord. Under such conditions, contraction time of the fast-twitch muscle remains unchanged. Also, cord transection of newborn animals does not affect the postnatal acceleration in contraction time of the fast-twitch muscle, while the postnatal changes of contraction of the slow-twitch muscle mimic those of the fast-twitch muscle. The contraction time of slow-twitch muscle fibers is correlated with the duration of after-hyperpolarization (AHP) in the motoneurons, which regulates the discharge frequency. The duration of AHP increases with age in the motoneurons innervating the slow-twitch muscle but not in those innervating the fast-twitch muscle. It is proposed that the maintenance of slow contractions in the slow-twitch muscle during muscle differentiation is due to the appearance of low frequency discharges in the innervating motoneurons as a result of the postnatal prolongation of the AHP. The implications and limitations of the proposed hypothesis are discussed.

Aging↗

PubMed1986

Modulation of calmodulin function and of Ca2+-induced smooth muscle contraction by the calmodulin antagonist, HT-74.

The relationship between the functions of calmodulin (CaM) and Ca2+-induced smooth muscle contraction was investigated using a newly synthesized CaM antagonist, 3-(2-benzothiazolyl)-4,5-dimethoxy-N-[3-(4- -phenylpiperidinyl)propyl]benzenesulfonamide (HT-74). We noted a selectivity of HT-74 for CaM, compared to other calcium-binding proteins and target enzymes of CaM. As HT-74 had no significant effect on the intensity of 8-anilino-1-naphthalene-sulfonic acid (ANS) fluorescence in the presence of the Ca2+-CaM complex, the HT-74-binding sites may differ from those of naphthalenesulfonamides and phenothiazines which decrease ANS fluorescence. The Ca2+ binding to CaM was inhibited significantly by 1.0 microM HT-74, in sharp contrast to phenothiazines and naphthalenesulfonamides which increase the extent of the Ca2+ binding to CaM. Increasing CaM concentrations reversed the HT-74-induced inhibition of CaM-dependent enzymes such as myosin light chain kinase and Ca2+-dependent cyclic nucleotide phosphodiesterase, with Ki values of 0.5 microM and 0.4 microM, respectively. In the presence of 0.3 microM HT-74, potassium-depolarized rabbit aortic strips pre-contracted with 0.3 mM CaCl2 relaxed, and this relaxation was completely reversed by the addition of an excess amount of CaCl2 (10 mM). This compound shifted the dose-response curve for CaCl2 to the right, in a competitive manner. However, HT-74 inhibited the phenylephrine-induced contraction elicited in Ca2+-free solution and the calcium ionophore A23187-induced contraction in the presence of calcium ion. Therefore, this agent affects intracellular actions of Ca2+ rather than membrane receptors or the influx of Ca2+. HT-74 is a CaM antagonist which binds to CaM in a manner different from that heretofore reported. It inhibits Ca2+ binding to CaM and produces a competitive inhibition of Ca2+-induced contractions of depolarized vascular smooth muscle.

2',3'-Cyclic-Nucleotide Phosphodiesterases↗

PubMed1999

Optimum length of muscle contraction.

OBJECTIVE: The purpose of this study was to develop a mathematical method to determine optimum muscle length and muscle stress based on the measurable physiological and biomechanical data. DESIGN: The values of optimum muscle length and muscle stress are investigated. BACKGROUND: Understanding the characteristics of muscle function in vivo is important for assisting the design of the tendon transfer and other rehabilitation procedures. In vivo determination of the physiological and anatomical parameters of muscle contraction is difficult but not impossible. Optimum muscle length and muscle stresses are important parameters for understanding muscle function. METHODS: A Cybex dynamometer was used to measure isometric elbow flexion torque in eight different joint positions in seven subjects. Then the optimization method was used to determine optimum muscle length and muscle stress of three major elbow flexors, the biceps brachii, the brachialis, and the brachioradialis based on the model and joint torque data. RESULTS: The calculated muscle stress for each subject was on average 109 N/cm(2), while the optimum muscle length for the biceps brachii, the brachialis, and the brachioradialis was on average 14.05, 6.53, 17.24 cm, respectively. The joint angles corresponding to these optimum muscle lengths are 110 degrees, 100 degrees and 50 degrees of elbow flexion, respectively. CONCLUSIONS: Optimum muscle length and muscle stress can be properly predicted using an analytical mathematical model along with an experimentally measured joint torque. RELEVANCE: The estimate of optimum muscle length is important for muscle modeling and tendon transfer surgery by taking advantage of length-tension relationship of individual muscles.

Adult↗

PubMed1992

Neutral endopeptidase modulates endothelin-1-induced airway smooth muscle contraction in guinea-pig trachea.

This study was designed to evaluate the role of neutral endopeptidase (NEP) in modulating the airway smooth muscle contraction induced by endothelin-1 in isolated segments of guinea-pig trachea. Endothelin-1 (10(-9)-10(-6) M) produced a concentration-dependent contraction that reached a maximum by 30 min. The NEP inhibitor leucine-thiorphan (10(-5) M) significantly increased the contractile response to endothelin-1. The addition of leucine-thiorphan to tracheal segments precontracted by 10(-9) and 10(-8) M endothelin-1 increased isometric tension by 181 +/- 65% (mean +/- 1 S.E.M.; P less than 0.05) and by 138 +/- 49% (P less than 0.05), respectively. In contrast, the kininase II inhibitor captopril and the peptidase inhibitors leupeptin and bestatin had no effect. Preincubation of endothelin-1 with 1 microgram recombinant human NEP decreased the contractile activity of endothelin-1 by 72 +/- 9%, whereas no effect was observed using heat-inactivated NEP. We conclude that NEP modulates endothelin-induced contraction of airway smooth muscle in the guinea-pig trachea.

Animals↗

PubMed1994

Regulation of vascular smooth muscle contraction: myosin light chain phosphorylation dependent and independent pathways.

Ca(2+)-dependent myosin light chain (MLC) phosphorylation is an important step in the initiation of smooth muscle contraction. However, MLC phosphorylation alone cannot account for all aspects of contractile regulation, suggesting the involvement of other elements. In this article we present evidence obtained from Triton X-100 detergent skinned and intact tissue which demonstrates that vascular smooth muscle contraction can be initiated by a Ca(2+)-dependent mechanism that does not require prior MLC phosphorylation. We show that Ca2+ can initiate contractions supported by cytidine triphosphate (CTP) and that these contractions are inhibited by calmodulin antagonists, suggesting a Ca(2+)-calmodulin dependence of force distinct from that for MLC phosphorylation. Evidence is presented to demonstrate that carotid medial fibers contain a mitogen-activated protein (MAP) kinase which is activated by Ca2+ and may catalyze caldesmon phosphorylation. Based in part on our results and those of other investigators, we propose that direct Ca(2+)-calmodulin binding to caldesmon or phosphorylation of caldesmon by a Ca(2+)-dependent MAP kinase disinhibits caldesmon. Disinhibition of caldesmon allows an inherent basal level of actin-activated myosin ATPase activity to be expressed. The result is the slow development of force.

Animals↗

PubMed1986

Afferent projections to human tibialis anterior motor units active at various levels of muscle contraction.

The synaptic efficacy of muscle and cutaneous afferents on single tibialis anterior motoneurones in man was derived from changes in the firing probability of single, voluntarily activated, motor units in response to electrical stimulation of peripheral nerves or skin. The motor units were recorded with a Macro EMG electrode. The Macro motor unit potential (Macro MUP) recorded with this electrode reflects the electrical activity of all of the muscle fibres in a single motor unit. The amplitude of the Macro MUP is positively correlated with the recruitment threshold of the unit. Motor units with different Macro MUP amplitudes were examined at approximately the same level of voluntary contraction (less than 20% of maximum). The synaptic efficacy of muscle and cutaneous afferents was similar for units with small and with large Macro MUP amplitudes. Single motor units were examined at several different levels of muscle contraction. There was no consistent change in the facilitation from muscle afferents but there was less facilitation from cutaneous afferents during stronger contractions. This was not simply a consequence of the units faster firing rate. It is concluded that, with increasing voluntary drive to tibialis anterior motoneurones in man, there is a reduction in transmission in the pathways from cutaneous afferents to tibialis anterior motoneurones. There is no evidence that low and high threshold units (judging from their Macro MUP amplitudes) have different afferent connections.

Action Potentials↗

PubMed1991

Perinatal changes in bombesin-stimulated muscle contraction in rabbit stomach and colon.

Bombesin and its mammalian homologue, gastrin-releasing peptide, stimulate smooth muscle contraction and may promote the growth of gastrointestinal tissues. Isometric contraction of strips from circular muscle of the gastric fundus and longitudinal muscle of the distal colon were used to compare changes in the response to bombesin in newborn and weanling rabbits. There was an age-related qualitative change in gastric muscle from biphasic contractions including phasic and tonic components in the newborn to phasic contractions alone in the weanling. The colon contractions were tonic at both ages. In both tissues there was an age-related fivefold increase in stress in response to maximally effective concentrations of bethanechol (P less than 0.05). In contrast, in the stomach age-related decreases in the response to maximally effective concentrations of bombesin were observed, from 2930 +/- 179 mN/cm2 (98% of the maximal response to bethanechol) in the newborn to 565 +/- 81 mN/cm2 (4% of the maximal response to bethanechol) in the weanling (P less than 0.005). In the colon, a twofold increase in response to bombesin was observed, from 446 +/- 59 mN/cm2 (82% of the response to bethanechol) in the newborn to 862 +/- 11 mN/cm2 (29% of the response to bethanechol) in the weanling (P less than 0.05). No age-related changes were observed in the potency of bombesin in either tissue. Neither atropine nor tetrodotoxin altered the contractions in either tissue, suggesting that bombesin interacted directly with myocytes. There was three times as much bombesinlike immunoreactivity in the stomach compared with the colon, but no age-related changes in either tissue. In summary, by the age of weanling the stomach lost the tonic component of contraction and 80% of the efficacy of bombesin-stimulated phasic contraction that had been present in the newborn. The loss of efficacy, absolute in the stomach and relative to bethanechol in the colon, suggest that bombesin may be most important in stimulating motility in the neonatal period.

Animals↗

PubMed2004

Aftereffects of mechanical vibration and muscle contraction on limb position-sense.

Mechanical vibration (MV) of a muscle causes position-sense errors during and after application. Isometric muscle contraction at a shorter (hold-short conditioning) or longer (hold-long conditioning) length causes limb position-sense errors after the muscle returns to its intermediate length by means of intrafusal muscle thixotropy. However, it is unclear whether MV enhances these thixotropic position-sense errors. We studied the after-effects of MV on position-sense errors induced by hold-short and hold-long conditioning in the biceps of 12 healthy men. After hold-short conditioning, subjects perceived that the conditioned forearm was placed in a more extended position than occurred in reality; after hold-long conditioning, a more flexed position was perceived. Use of MV with hold-short or hold-long conditioning enhanced both errors, which were most obvious at 100 HZ. These results suggest that MV and muscle conditioning work together efficiently to develop intrafusal muscle thixotropy. MV combined with hold-long conditioning may alleviate thixotropically increased muscle stiffness, such as in spastic hypertonia.

Adult↗

PubMed2001

Response to high-intensity eccentric muscle contractions in persons with myopathic disease.

Although the response to intense eccentric muscle contractions is well described in normal subjects, concern exists about possible untoward effects in persons with myopathic diseases. We investigated 14 subjects with slowly progressive muscular dystrophies including myotonic muscular dystrophy (n = 9), facioscapulohumeral dystrophy (n = 2), limb-girdle syndrome (n = 2), and Becker muscular dystrophy (n = 1). Control subjects consisted of 18 able-bodied persons. Subjects performed two sets of eight maximal-effort eccentric repetitions of the elbow flexors, with measurement of maximal concentric strength, serum creatine kinase, resting and flexed arm angle, arm circumference, and soreness at days 0, 3, and 7. Although the myopathic group had less initial strength, both groups demonstrated a similar response to the protocol over 7 days. Both groups had a significant rise in serum creatine kinase, which was still elevated at 7 days (P < 0.05). The control group demonstrated a slightly greater injury response in terms of soreness, resting and flexed arm angles, and arm swelling. Both groups of subjects appeared to respond similarly to an acute bout of eccentric contractions. However, the potential long-term effects of this type of exercise in persons with myopathic diseases remains unknown.

Adult↗

PubMed1975

In vivo recovery of muscle contraction after alpha-bungarotoxin binding.

Acetylcholine receptors were inactivated in vivo at the mouse neuromuscular junction using alpha-bungarotoxin (alpha-BTX). It was found that neurally produced muscle contraction recovered within 4-8 days (halftime similar to 3 days). Actinomycin D interfered with this recovery, but did not affect normal nerve-stimulated muscle contraction. If the response was initially eliminated by [125-I]alpha-BTX and the end plates examined by EM autoradiography, no evidence of mass internalization of bound radioactivity during recovery was seen. The fine structure of the end plates and muscle was unaltered during the post-alpha-BTX recovery period.

Acetylcholine↗

PubMed1996

Microtubules regulate pulmonary vascular smooth muscle contraction.

Microtubules are ubiquitous in eukaryotic cells. However, the role of microtubules in the mechanisms of pulmonary vascular smooth contraction has not previously been described. The purpose of this study was to examine the effect of microtubular inhibition (vinblastine) on the following mechanisms of pulmonary vascular smooth muscle contraction in rats using isolated pulmonary artery rings: (1) receptor-independent, calcium-dependent contraction via smooth muscle cell depolarization (response to KCl); (2) receptor-dependent, calcium-dependent contraction via alpha 1-adrenergic receptor stimulation (response to phenylephrine, PE); (3) receptor-dependent, calcium-independent contraction via thromboxane A2 receptor stimulation (response to the thromboxane mimetic, U-46619). Rats were studied 4 days after administration of vinblastine (750 micrograms/kg i.v.). Concentration-response curves were generated for KCl (5 mM to 100 mM) and for PE and U-46619 (10(-9) to 10(-4) M) (n = 8 rings/4 rats per group). Saline injected rats were controls. Pulmonary vascular smooth muscle contraction by calcium-dependent and -independent mechanisms was significantly increased following microtubular inhibition. These findings suggest that microtubules have an important role in the response of pulmonary vascular smooth muscle to vasoconstricting agonists.

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

PubMed1996

A simple, non-invasive and inexpensive method for evaluating the displacement of local tissue surfaces: from vascular changes to muscle contraction.

This paper describes a non-invasive, very inexpensive method of estimating tissue displacements of various origin that is easy and fast to set up. This technique utilizes an inductive proximity sensor (IPS), which is a non-contact length transducer measuring the distance between its probe and a metal target. Its working principle is based on the electromagnetic coupling originating between the sensor probe, a source of high-frequency magnetic field, and the metal target where parasitic currents take place. The linear working range of the IPS model used here is 0.1 to 6 mm probe-target distance, its resolution is about 2 microns. The IPS has been employed on rabbits and humans to measure the displacement of a target glued to the skin of various body areas with respect to the fixed probe of the sensor. Its high resolution, together with an extensive working range, allows the evaluation of numerous physiological events which produce displacements ranging from 2 microns -- to 9 mm, reflecting either tissue volume changes or movements. In particular, an interesting application is to monitor, through volume variations, the extent and the time course of local vascular modifications induced by manoeuvres which elicit changes in vasomotor tone; vascular filling, tissue swelling etc. Therefore, this measure may be considered a 'surface plethysmography' record. In addition, the contractions of skeletal muscles, under either isotonic or isometric conditions, can be estimated through this sensor. This system may therefore find applications for research purposes and practical demonstrations to students.

Animals↗

PubMed1989

Information transfer in the regulation of striated muscle contraction.

A brief review of current views on the regulation of striated muscle contraction is presented with emphasis on the transmission of the effect of calcium binding to the N-terminal domain of troponin C to the other components of the regulatory system. Results of recent work on chemical modification of troponin C as well as studies on crosslinking among thin filament components are discussed.

Animals↗

PubMed1998

A large and distinct rotation of the myosin light chain domain occurs upon muscle contraction.

For more than 30 years, the fundamental goal in molecular motility has been to resolve force-generating motor protein structural changes. Although low-resolution structural studies have provided evidence for force-generating myosin rotations upon muscle activation, these studies did not resolve structural states of myosin in contracting muscle. Using electron paramagnetic resonance, we observed two distinct orientations of a spin label attached specifically to a single site on the light chain domain of myosin in relaxed scallop muscle fibers. The two probe orientations, separated by a 36 degrees +/- 5 degrees axial rotation, did not change upon muscle activation, but the distribution between them changed substantially, indicating that a fraction (17% +/- 2%) of myosin heads undergoes a large (at least 30 degrees) axial rotation of the myosin light chain domain upon force generation and muscle contraction. The resulting model helps explain why this observation has remained so elusive and provides insight into the mechanisms by which motor protein structural transitions drive molecular motility.

Animals↗