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Responses of rat isolated intestinal segments to stimulation of perivascular mesenteric nerve fibres.

1. Isolated preparations of segments of rat jejunum were set up for isotonic recording of the activity of the longitudinal smooth muscle and the extrinsic nerve supply running with the mesenteric blood vessels was stimulated for 30-s periods at 0.5-20 Hz. 2. Contractions were regularly elicited during periods of stimulation at 0.5-2 Hz, and were usually elicited during stimulation at 5 Hz, but relaxations were usually elicited during stimulation at 10 and 20 Hz. On cessation of the period of stimulation, a secondary contraction occurred in most preparations regardless of whether the primary response during the period of stimulation had been a contraction or a relaxation. 3. Tetrodotoxin (0.9 microM) abolished responses during periods of stimulation and the secondary contraction. 4. The relaxations were mimicked by noradrenaline and were abolished by blockade of alpha- plus beta-adrenoceptors but were not affected by hexamethonium, indicating that they are attributable to stimulation of postganglionic noradrenergic fibres. 5. The contractions were mimicked by acetylcholine and were abolished by atropine and hexamethonium, suggesting that they are attributable to stimulation of preganglionic cholinergic nerves: they were also abolished in the presence of capsaicin, indicating that sensory neuropeptide-containing nerve fibres may be involved. 6. The secondary contraction that usually occurred on cessation of stimulation was still present after blockade by atropine or capsaicin of the contractions occurring during stimulation. It was also present after blockade by propranolol and phentolamine of the relaxations occurring during stimulation, and was not affected by indomethacin.

Animals↗

Pharmacological characterization of the histamine receptor in the isolated muscularis mucosae of the guinea-pig oesophagus.

To characterize the histamine receptors in the muscularis mucosae, the isotonic responsiveness of the isolated muscularis mucosae of the guinea-pig oesophagus to histamine receptor agonists and antagonists was examined in vitro. Histamine (0.1-100 microM) produced a concentration-dependent contraction of the muscularis mucosae (EC50 = 1.6 +/- 0.2 microM). The contractions were rapid in onset, sustained, reversible by washing and the preparation did not show tachyphylaxis. 2-Methylhistamine (2-MH), 2-pyridylethylamine (PEA) and 4-methylhistamine (4-MH) produced similar sustained contractions of the muscularis mucosae. The order of sensitivity was histamine greater than 2-MH greater than PEA greater than 4-MH. Impromidine (10-300 microM) and dimaprit (10-300 microM) caused no response in this tissue. The contractile responses to histamine, 2-MH, and PEA were competitively antagonized by diphenhydramine, and the pA2 values were almost the same (approximately 8.1). Cimetidine (100 microM) could not modify the contractile response to these agonists. The contractile response to histamine was slightly inhibited by tetrodotoxin (0.3 microM), atropine (1 microM), indomethacin (0.1-3 microM) or aspirin (30-300 microM), and the EC50 value was increased about 2-6 times by these drugs. When the preparation was incubated in Tyrode solution containing various calcium concentrations (0, 0.45, 0.9 and 1.8 mM), the concentration-response curve to histamine was shifted to the right and downward; the effect was inversely dependent on the calcium concentration, and in a calcium-free medium the response to histamine was abolished. Verapamil (1-10 microM) partially inhibited the contractile response to histamine. 7 The present results indicate that the contraction of the guinea-pig oesophageal muscularis mucosae to histamine is mediated mainly by a direct action on the smooth muscle and partly by indirect actions via the stimulation of either endogenous prostaglandin biosynthesis or intramural cholinergic nerves. The histamine receptors responsible for contractions of this tissue are probably mainly of the H,- subtype with H2-receptors having a negligible role.

Animals↗

The effect of ouabain on the guinea pig ileum longitudinal smooth muscle: 2. Intracellular levels of Ca, Na, K, and Mg during the ouabain response and the dependence of the response on extracellular Ca.

Isotonic Tris-HCl containing 10 mM LaCl3 at 4 degrees C effectively removed extracellular ions in 30 min while preventing loss of intracellular ions. Intracellular Ca and Na increased during the contraction in the presence of 10 mM ouabain and then decreased during relaxation. Intracellular Na increased again during the latter part of the relaxation phase when K loss became apparent. Mg levels remained essentially constant. Ouabain responses were rapidly lost in Ca-free medium indicating that they were dependent on extracellular Ca. A 5.5-fold increase in the normal levels of extracellular K did not reduce the contraction to a submaximal dose of ouabain. A full phasic response to high K (60 mM) was observed after a 10-min exposure of the tissue to ouabain, at which time the ouabain response had returned to basal tension. The contraction to ouabain appears to be dissociated from inhibition of the Na,K-ATPase at the K site. The changes in intracellular ions indicated that ouabain contracted the muscle by increasing the plasma membrane permeability to Ca and Na and later decreased the K and Na concentration gradients, probably by inhibition of the Na,K-ATPase.

Adenosine Triphosphatases↗

CONTRACTILE PROPERTIES OF A HIGH-FREQUENCY MUSCLE FROM A CRUSTACEAN - CONTRACTION KINETICS

1. The flagella (small appendages on the maxillipeds) of the crab Carcinus maenas beat regularly when active at about 10 Hz (15 °C). The beat of a flagellum is due to contraction of a single small muscle, the flagellum abductor (FA). The optimal stimulus frequency for tetanic contraction of the FA was about 200 Hz. When the muscle was stimulated at 10 Hz with paired stimuli per cycle, the interstimulus interval that maximized peak force was 2­4 ms, which corresponded well to the interspike intervals within bursts recorded from motor axons during normal beating. 2. Contraction of the isolated FA showed pronounced neuromuscular facilitation and many stimuli were needed to activate the muscle fully. The dependence on facilitation in isolated muscles appeared to be greater than that in vivo. It is suggested that neuromodulators in the blood of the crab enhance neuromuscular transmission and reduce the dependency on facilitation in intact animals. 3. The FA had a narrow length­tension curve. Tetanic tension became vanishingly small at muscle lengths less than about 90 % of the maximum in vivo length. The maximum length change of the muscle during in vivo contraction was about 5 %. 4. The maximum isometric force of the FA was low (about 6 N cm-2) but its shortening velocity was high. Vm, the maximum shortening velocity determined from isotonic shortening, was 4.0 muscle lengths s-1; V0, the maximum shortening velocity from slack test measurements, was about 8 lengths s-1. 5. The structure and physiology of the FA are compared with those of locust flight muscle, chosen because it too is a muscle capable of long-duration, high-frequency performance.

Journal Article↗

[Functional status of the circulatory system in during dynamics of treating arterial hypertension in patients with vibration disease].

AIM: To develop medicinal approaches to correction of hemodynamic disturbances in vibration disease (VD) associated with arterial hypertension. MATERIAL AND METHODS: The study compared hypotensive and hypodynamic efficiency of amlodipin, diltiazem, enalapril, perindopril and indapamide in 74 VD patients with arterial hypertension (SAP 140-179 mm Hg, DAP 90-109 mm Hg; mean age 54.8 years, mean exposure to vibration 26.8 years). Before and after the treatment course the patients were examined with ultrasound by the following parameters: left ventricular contractility, left and right ventricular diastolic function, left atrial function. RESULTS: Amlodipin reduced left ventricular volume both in systole and diastole as well as maximal intramyocardial tension without significant change in contractility, raised a contraction reserve of the left atrium, improved ventricular relaxation. Diltiazem potentiates contractility of the left ventricle and atrium without marked impact on relaxation and tension of the myocardium. Enalapril and perindopril cause positive hemodynamic shifts. Perindopril vs enalapril was more effective in easing intramyocardial systolic tension of the left ventricular wall and in improving the diastolic function. Indapamid vs calcium antagonists and ACE inhibitors had a weaker effect on arterial pressure, no significant effect on left ventricular contraction, peripheral hemodynamics. Left atrial function was hyperactive. This was observed also in response to the other drugs. CONCLUSION: Amlodipin produced in patients with VD and AH more positive hemodynamic effects, reduced isotonic hyperfunction of the left ventricle, improves diastolic function of the ventricles. Amlodipin and perindopril are more promising prognostically in relation to reduction of left ventricular myocardial mass.

Antihypertensive Agents↗

Muscle force arises by actin filament rotation and torque in the Z-filaments.

Actin filament rotation in skeletal muscle is studied by a mechanical model that simulates structure and tension. The four anchoring Z-filaments are twisted around and change the structure of the Z-lattice. The "small square" without twist represents the resting stage of muscle. Torque causes contraction by clockwise rotation (as seen from the Z-band), drilling into the A-band and transition of the "small square" to "basket weave" by increasing the twist and decreasing the torque. Release decreases the torque ("force-depression") by passive clockwise rotation. Stretch causes increased torque ("stretch activation") by passive counterclockwise rotation. Torque arises during Ca(2+)-activation by a conformational change in the highly charged coiled-coils: The four alpha-actinin Z-filaments generate strong torque for the isometric tension. Quick release experiments show that less than one rotation reduces this torque to zero. The 5-12 rotations necessary for isotonic shortening result from torque-generation in the two long tropomyosin coiled-coils. Myosin controls the velocity of active and passive rotations.

Actinin↗

Mechanisms of mechanical strain memory in airway smooth muscle.

We evaluated the hypothesis that mechanical deformation of airway smooth muscle induces structural remodeling of airway smooth muscle cells, thereby modulating mechanical performance in subsequent contractions. This hypothesis implied that past experience of mechanical deformation was retained (or "memorized") as structural changes in airway smooth muscle cells, which modulated the cell's subsequent contractile responses. We termed this phenomenon mechanical strain memory. Preshortening has been found to induce attenuation of both force and isotonic shortening velocity in cholinergic receptor-activated airway smooth muscle. Rapid stretching of cholinergic receptor-activated airway smooth muscle from an initial length to a final length resulted in post-stretch force and myosin light chain phosphorylation that correlated significantly with initial length. Thus post-stretch muscle strips appeared to retain memory of the initial length prior to rapid stretch (mechanical strain memory). Cytoskeletal recruitment of actin- and integrin-binding proteins and Erk 1/2 MAPK appeared to be important mechanisms of mechanical strain memory. Sinusoidal length oscillation led to force attenuation during oscillation and in subsequent contractions in intact airway smooth muscle, and p38 MAPK appeared to be an important mechanism. In contrast, application of local mechanical strain to cultured airway smooth muscle cells induced local actin polymerization and cytoskeletal stiffening. It is conceivable that deep inspiration-induced bronchoprotection may be a manifestation of mechanical strain memory such that mechanical deformation from past breathing cycles modulated the mechanical performance of airway smooth muscle in subsequent cycles in a continuous and dynamic manner.

Adaptation, Physiological↗

The undamped and damped series elastic components of a vascular smooth muscle.

Small arterial resistance vessels (internal diameter about 175 micrometer) have been mounted on a myograph that enabled their wall tension, T, and internal circumference, L, to be measured and controlled with a time resolution of about 4 ms. Maximally activated vessels were subjected to isometric releases (step changes in L) and isotonic releases (step changes in T) of varying extents and at two different temperatures (27 degree C and 37 degree C). The recovery from an isometric release was monotonic and did not include the two phases seen in skeletal muscles. The isotonic release response did, however, contain a velocity transient lasting about 150 ms: the velocity immediately after the release was about six times the steady shortening velocity. The form of both the isometric and isotonic release responses and their dependence on the extent of release can be explained in terms of a modified Hill model in which the "series elastic component" (SEC) is replaced by the series combination of an undamped-SED (that is, an undamped elastic element) and a damped-SEC (a Voigt element). Although the initial response to both types of release was independent of temperature, all stages of subsequent responses were temperature dependent, with Q10's in the range 1.5 - 2.0. The results suggest that the responses to isotonic and isometric releases may in part be due to active processes.

Animals↗

Characterization of neuropathic bladder smooth muscle cells in culture.

PURPOSE: Clinically bladder cells used in tissue engineering techniques will come from neuropathic bladders and not normal bladders. We determined if neuropathic bladder smooth muscle (SM) cells (SMCs) retain functional differences when cultured in vitro. MATERIALS AND METHODS: Primary cultures of SMCs were established from patients with a neuropathic bladder (5) and a normal bladder (5). Expression of alpha-SM actin and SM myosin heavy chain was determined using immunocytochemical staining and Western blot analysis. Baseline cell proliferation and the mitogenic response to angiotensin II was assessed by cell counting and cell viability assays. Cell contractility was determined for normal and neuropathic SMCs using an in vitro collagen lattice assay. Cell adherence was measured assessed using partial and complete trypsinization assays. RESULTS: Normal and neuropathic SMCs showed similar morphology in culture, and similar patterns of alpha-SM actin and SM myosin expression. Following 10 days of plating under optimal growth conditions the number of neuropathic SMCs was 170% more than normal SMCs. In response to angiotensin II neuropathic SMCs reached 54% of maximal growth capacity as opposed to 30% for normal SMCs (p <0.01). Neuropathic SMCs contracted significantly less in 10% serum and calcium ionophore (p <0.05), as determined by in vitro contractility assays. Neuropathic SMCs had 19% and 30% less adherent cells than normal SMCs (p <0.01) following isotonic solution washes and trypsinization, respectively. CONCLUSIONS: These results demonstrate that cultured neuropathic bladder SMCs possess and maintain different characteristics than normal SMCs in vitro. The potential clinical implications of using these cells in conjunction with tissue engineering techniques for the promotion of bladder regeneration requires further investigation.

Cells, Cultured↗

Electron microscopy of histamine-induced contraction of the in vitro swine coronary artery.

Dose-dependent contractions of the in vitro swine coronary artery were induced by application of histamine and acetylcholine, but not of angiotensin II, ergonovine, noradrenaline, prostaglandin F2 alpha and serotonin. Ultrastructural changes especially of the tunica intima during the contractions were observed at 2, 5 and 30 min after application of histamine and acetylcholine. The intimal gutter spirally running along the longitudinal axis of the vessel was obscured, and the intimal surface became extensively indented. Exclusively in the histamine-treated samples, the increase in number and size of the intracellular vacuoles and the dilation of the intercellular clefts to the extent of the intercellular vacuoles were observed in the endothelium. Moreover, the enhancement of the endothelial permeability was indicated by the marker experiments using horseradish peroxidase. Such endothelial cell damages and the enhancement of the endothelial permeability may amplify the coronary artery contraction.

Acetylcholine↗

Chronic calcium channel blockade prevents the progression of myocardial contractile and electrical dysfunction in the cardiomyopathic Syrian hamster.

The programmed onset of myocardial dysfunction and its progression to congestive heart failure in the cardiomyopathic Syrian hamster is hallmarked by alterations in myocellular calcium regulation. To determine whether calcium channel blockade is effective in halting the progressive depression of myocardial contractile performance in this animal model of congestive heart failure, oral verapamil therapy was instituted at 50 days of age, and treatment continued for various durations until the time of study at either 150 or 250 days of age. Left ventricular papillary muscle isometric and isotonic performance, as well as transmembrane electrical characteristics, was depressed in diseased hamsters at 150 days of age and deteriorated further by 250 days of age. These changes were evidenced by prolongation of contraction duration, a marked depression in the load-velocity relation, and a significant prolongation in the repolarization phase of the transmembrane action potential. Myocardial functional and electrical alterations associated with the progression of life in myopathic hamsters were completely halted by verapamil therapy that was continuous from 50 days after birth until death by diastolic arrest, at 150 or 250 days of age. However, premature termination of verapamil treatment before death resulted in a progressive renewal of the functional and electrical alterations for the duration of drug termination. It is concluded that the pathological changes seen during the lifetime of the cardiomyopathic hamster can be prevented by continuous calcium channel blockade and that intermediate prevention can be attained by protracted verapamil therapy. Thus, chronic verapamil therapy may be a useful adjunct in the prevention of human congestive heart failure of similar etiology.

Animals↗

The stiffness under isotonic releases during a twitch of a frog muscle fibre.

These investigations were undertaken to elucidate whether the difference between the time courses of the isometric tension and the velocity of shortening upon releases to a given isotonic load various times during a twitch is due to a change in the kinetics of the individual cross-bridges, i.e. in the rate constants governing the transitions between the various strongly bound cross-bridge states. Single skeletal muscle fibres from the frog (Rana temporaria, sarcomere length 2.3 microns, temperature 4-6 degrees C) were stimulated to produce isometric twitches and released after various delays to shorten isotonically against finite loads (P). Force, stiffness (K, measured by longitudinal oscillation 0.3-0.8 nm per half-sarcomere peak-to-peak, 2 kHz), and change in length were measured simultaneously. The velocity of shortening (V) was estimated as the quasi steady-state value, i.e. the value attained after the initial oscillatory transient response to the change in length preceding the isotonic shortening. A reduction in P led to a reduction in K, however, the relative reduction in K was not as great as the relative reduction in P. Thus, when extrapolated to P = O, K was about one third of its peak isometric value. Under the assumption that K is to the first order of approximation proportional to the number of strongly bound cross-bridges, P was divided by K to give an estimate of the average load per cross-bridge yo = P/K.V showed an almost linear dependence on yo (correlation coefficients 0.95-0.998). The slope and intercept of this relation (regression analysis) showed no dependence on the delay after stimulation of the release to the isotonic load. The results can be explained if the kinetic properties of the individual strongly bound cross-bridges are unaffected by the level of activation which changes during the course of the contraction.

Animals↗

Immediate mechanical reactions of isolated human detrusor muscle on x-irradiation.

The mechanical activity (isotonic registration) of isolated detrusor (smooth) muscle preparations from the human urinary bladder (surgical material) is investigated. The preparations reacted on X-irradiation (50 kV) with an immediate dose-dependent tonic contraction at threshold-doses of 100 to 1000 R (1 to 3 kR/min). The contraction was either reversible, semireversible or persistent, and the basal muscle tone was elevated. A tachyphylactic reaction was observed. The phasic activity of the preparations was temporarily increased, then inhibited after irradiation. The radiation-induced changes of the human and guinea-pig detrusor muscle are compared and their possible relation to the functional disturbances of the urinary bladder after radiotherapy is discussed.

Animals↗

Experimental muscle pain changes motor control strategies in dynamic contractions.

This study investigated the effect of muscle pain on muscle activation strategies during dynamic exercises. Ten healthy volunteers performed cyclic elbow flexion/extension movements at maximum speed for 2 min after injection of (1) hypertonic (painful) saline in the biceps brachii, (2) hypertonic saline in both biceps brachii and triceps brachii, and (3) isotonic (nonpainful) saline in the biceps brachii muscle. Surface electromyographic (EMG) signals were collected from the upper trapezius, biceps brachii, triceps brachii, and brachioradialis muscles (to estimate EMG amplitude) and with an electrode arrays from biceps brachii (to estimate muscle fiber conduction velocity [CV]). In all conditions, the acceleration of the movement decreased throughout the exercise, and kinematic parameters were not altered by pain. With respect to the control condition, pain induced a decrease of the biceps brachii (mean +/- SE, -23+/-4%) and brachioradialis (-10+/-0.4%) integrated EMG (IEMG) in the beginning of the exercise, and an increase (45+/-3.5%) of the upper trapezius IEMG at all time points during the exercise. The biceps brachii IEMG decreased over time during the nonpainful exercises (-11+/-0.6%) while it remained constant in the painful condition. Biceps brachii CV decreased during painful conditions (-12.8+/-2.2%) while it remained constant during the nonpainful condition. In conclusion, muscle pain changes the motor control strategy to sustain the required dynamic task both in the relative contribution between synergistic muscles and in the motor unit activation within the painful muscle. Such a changed motor strategy may be highly relevant in models of occupational musculoskeletal pain conditions.

Adaptation, Physiological↗

Release of prostaglandins E and F from rat uterine strips incubated in depolarizing and hyperpolarizing solutions.

The purpose of this study was investigate the effect of depolarizing (high K+, procaine 5 mM, high K+-Ca2+ free) or hyperpolarizing (K+ free) solutions on the generation and release of prostaglandins (PGs) E and F from the rat uterus. The output of PGs in the bathing solution was measured by bioassay. Results are expressed as ng/g dry weight/min. Under basal conditions in Krebs Ringer Bicarbonate medium, the total output of PGs (E + F) was 12.8 +/- 0.9, 20.5 +/- 2.5 and 39.1 +/- 4.3 ng/dw/min at 0-60, 60-80, 80-100 min intervals respectively, the major proportion being PGF like material in all cases. When the strips were bathed in depolarizing solution the total output increased significantly PGE being the most augmented. However, when K+ free solution was used there was a marked reduction in prostaglandins release as compared with basal conditions or depolarizing media. These results can not be attributed to osmotic effect, muscular work or neurotransmitter release from nerve endings. A close relationship between the variations in membrane potential and prostaglandins release seems to indicate that both parameters might correlate well.

Animals↗