The Mechanical Properties of in Situ Canine Skeletal Muscle.

This study was undertaken to determine if fiber arrangement was responsible for differences in the whole muscle mechanical properties. Experiments were carried out in situ in blood perfused dog skeletal muscles at approximately normal body temperature between 36° and 38°C. The following mechanical relationships were studied using a pneumatic muscle lever to measure Tension (P), length (L) and dP/dt: and dL/dt with a high frequency oscillograph (500-1000 Hz): 1.) Length:Tension; 2.) Force:Velocity; and 3.) Stress:Strain of Series Elastic. Electron microscopy and fiber typing were done as adjunctive studies. Muscles were stimulated by direct nerve stimulation with 0.1msec stimuli at a rate of 1 impulse per second for twitch contractions, or in 200 msec bursts of 100 Hz 0.1 msec stimuli for brief tetanic contractions. The pennate short fibered gastrocnemius plantaris developed 1.0 kg/g of tension during brief tetanic stimulation, at optimal length (Lo) with full stimulus voltage, while the parallel long fibered semitendinosus developed 0.5 kg/g under the same conditions. The Length:Tension relationship for these two muscles was qualitatively similar but quantitatively different. The Force:Velocity relationship (ΔL/L0 vs. P/P0) for both muscles were also qualitatively similar and could be described by the previously proposed rectangular hyperbola but a better predicted fit to the observed data could be produced by adding a descending exponential function to the rectangular hyperbola. Unlike previous studies, the Stress:Strain properties of the series elastic component measured by quick release (ΔL/Li vs. ΔP/Po) were linear and gastrocnemius was 25 per cent higher than the semitendinosus. Overall, both muscles were found to have mechanical properties that differed little from the previously reported literature for amphibian, cardiac and small mammalian muscles studied by others in vitro. The major differences that we found were in the shapes of the force:velocity curve of the contractile component, and the Stress:Strain curve of series elastic component. Equations and explanations for these differences are devised and presented.

Positive inotropism in mammalian skeletal muscle in vitro during and after fatigue.

We tested the hypothesis that positive inotropic factors decrease fatigue and improve recovery from fatigue in mammalian skeletal muscle in vitro. To induce fatigue, we stimulated mouse soleus and extensor digitorum longus (EDL) to perform isometric tetanic contractions (50 impulses x s(-1) for 0.5 s) at 6 contractions x min(-1) for 60 min in soleus and 3 contractions x min(-1) for 20 min in EDL. Muscles were submerged in Krebs-Henseleit bicarbonate solution (Krebs) at 27 degrees C gassed with 95% nitrogen - 5% carbon dioxide (anoxia). Before and for 67 min after the fatigue period, muscles contracted at 0.6 contractions x min(-1) in 95% oxygen - 5% carbon dioxide (hyperoxia). We added a permeable cAMP analog (N6, 2'-O-dibutyryladenosine 3':5'-cyclic monophosphate at 10(-3) mol x L(-1) (dcAMP)), caffeine (2 x 10(-3) mol x L(-1), or Krebs as vehicle control at 25 min before, during, or at the end of the fatigue period. In soleus and EDL, both challenges added before fatigue significantly increased developed force but only caffeine increased developed force when added during the fatigue period. At the end of fatigue, the decrease in force in challenged muscles was equal to or greater than in controls so that the force remaining was the same or less than in controls. EDL challenged with dcAMP or caffeine at any time recovered more force than controls. In soleus, caffeine improved recovery except when added before fatigue. With dcAMP added to soleus, recovery was better after challenges at 10 min and the end of the fatigue period. Thus, increased intracellular concentrations of cAMP and (or) Ca2+ did not decrease fatigue in either muscle but improved recovery from fatigue in EDL and, in some conditions, in soleus.

Increased cAMP as a positive inotropic factor for mammalian skeletal muscle in vitro.

To test the hypothesis that an increased cAMP concentration improves skeletal muscle force development, we stimulated mouse soleus and extensor digitorum longus (EDL) in the presence of isoproterenol (1 x 10(-5) mol.L-1), a beta-adrenergic agonist, or N6,2'-O-dibutyryladenosine 3':5'-cyclic monophosphate (dcAMP) (1 x 10(-3) mol.L-1), a membrane-permeable cAMP analogue. Drugs used in the challenges were dissolved in Krebs-Henseleit bicarbonate buffer (Krebs) at 27 degrees C and gassed with 95% O2 - 5% CO2. Stimulation at 50 impulses.s-1 for 0.5 s produced an isometric tetanic contraction. Over 25 min of contractions at 0.6 contractions.min-1, developed force increased significantly with the addition of isoproterenol (soleus, 2.5% +/- 1.1%; EDL, 13.8% +/- 2.0%) or dcAMP (soleus, 2.3% +/- 0.5%; EDL, 10.9% +/- 1.9%) as compared with vehicle controls (cont) with Krebs added (soleus, 0.0% +/- 0.2%; EDL, -2.5% +/- 0.7%). To investigate the role of Ca2+ availability, we amplified or attenuated sarcolemmal L-type Ca2+ channels with Bay K 8644 (Bay K) (5.6 x 10(-6) mol.L-1) or diltiazem hydrochloride (dilt) (10(-4) mol.L-1), respectively. Ca2+ release from the sarcoplasmic reticulum was increased with caffeine (2 x 10(-3) mol.L-1) or decreased with dantrolene sodium (dant) (4.2 x 10(-7) mol.L-1). With Ca2+availability modified, dcAMP addition in soleus significantly increased force development above control (cont, 2.3% +/- 0.4%; Bay K, 4.0% +/- 1.0%; dilt, 52.3% +/- 3.6%; caffeine, 2.3% +/- 0.7%; dant, 6.0% +/- 2.0%; dilt + dant, 55.0% +/- 23.0%). In EDL, the addition of dcAMP also increased force development above control (cont, 13.7% +/- 1.9%; Bay K, 17.0% +/- 4.0%; dilt, 170.0% +/- 40.0%; caffeine, 23.0% +/- 4.0%; dant, 72.0% +/- 10.0%; dilt + dant, 54.0% +/- 14.0%). Thus, a positive inotropic effect of cAMP existed in both fast- and slow-twitch mammalian skeletal muscle with both normal and altered Ca2+ flux into the sarcoplasm.

Inotropic effects on mammalian skeletal muscle change with contraction frequency.

Over the last decade, we have attempted to determine if mammalian skeletal muscle's steady-level force development as established by mechanical and stimulation parameters can be increased or decreased by physiological signals. In these experiments, nitric oxide (NO), endothelin-1 (ET-1), adenosine (Ado), and beta-adrenergic agonists (beta) modified force production in the soleus and (or) the extensor digitorum longus (EDL) of the mouse. NO and beta increased the force produced by 0.5-s tetanic contractions at 0.6 contractions/min in both muscles. While EDL did not respond to either Ado or ET-1, the developed force of the soleus was amplified by Ado but attenuated by ET-1. Increased cAMP analogue concentrations amplified developed force in both muscles, but a cGMP analogue had no effect on either muscle. Following an increase in the contraction frequency of the soleus, the increased force in response to NO disappeared, as did the decreased force to ET-1. The increase in force due to a cAMP analogue disappeared during fatigue but reappeared quickly during recovery. Thus, steady-level developed force can be modified by a number of substances that can be released from locations in the body or muscle. The response to a given compound is determined by a complex interaction of metabolic and intracellular signals on the force-generating cascade.

Anterior coracoscapular ligament and suprascapular nerve entrapment.

A reduction in the height of the suprascapular foramen may predispose to entrapment of the suprascapular nerve. In this study, 16 of 27 cadavers (60%) demonstrated a heretofore unreported ligament located on the anterior aspect of the suprascapular foramen. In 11 of the 27 cadavers (41%), the ligament was observed bilaterally. The ligament decreased the foraminal height from the normative value of 5.6 +/- 0.4 to 2.3 +/- 0.4 mm (mean +/- SEM). Because this ligament, for which we propose the term anterior coracoscapular ligament (ACSL), substantially narrows the suprascapular foramen, it should be considered as a possible etiologic factor in suprascapular nerve entrapment.

The inotropic effect of nitric oxide on mammalian papillary muscle is dependent on the level of beta1-adrenergic stimulation.

We tested the hypothesis that nitric oxide has a positive inotropic effect on mammalian cardiac muscle contractility and that this effect sums with the positive inotropic effect of beta1-adrenergic agonists when both are present. Feline right ventricular papillary muscles were stimulated to contract isometrically at 0.2 Hz in Krebs-Henseleit bicarbonate buffer (KREBS) gassed with 95% O2 and 5% CO2 (26 degrees C; pH 7.34). The nitric oxide (NO) donor, S-nitroso-N-acetylpenicillamine (SNAP, 10(-5) M), and the membrane permeable cGMP analog 8-bromoguanosine-3',5'-cyclophosphate sodium (Br-cGMP, 10(-5) M), significantly increased developed force by 13.3+/-1.5% (n = 11) and 7.8+/-2.8% (n = 7), respectively. SNAP, at 10(-5) M, significantly increased the force developed by papillary muscle treated with 10(-11) M or 10(-9) M dobutamine hydrochloride (a beta1-adrenergic agonist) (n = 25, 11.3+/-2.9% and 10.0+/-3.6%, respectively) when compared with the addition of KREBS (n = 27, 2.6+/-0.9% and 5.5+/-0.9%), but the increase was less than predicted by the sum of inotropic effects of SNAP and dobutamine. SNAP at 10(-5) M did not change developed force in muscles treated with 10(-7) M dobutamine but it significantly decreased developed force in muscles challenged with 10(-5) M dobutamine (n = 18, 29.3+/-5.0%) when compared with KREBS (n = 10, 41.5+/-6.8%). Similarly, 10(-4) M 8-bromo-adenosine cyclic 3',5'-hydrogen phosphate monosodium (a membrane permeable cAMP analog) increased developed force 14.9+/-3.3% and the addition of 10(-5) M Br-cGMP to those muscles significantly reduced developed force by 3.5%+/-1.1% (n = 7). Thus, the positive inotropic effect of NO decreased and ultimately became an attenuation as the level of beta1-adrenergic stimulation increased due at least in part, to an interaction between the cAMP and cGMP second messenger pathways.

A1 receptor activation decreases fatigue in mammalian slow-twitch skeletal muscle in vitro.

To test the hypothesis that adenosine improves skeletal muscle cell function, we exposed curarized mouse soleus and extensor digitorum longus (EDL) to a range of concentrations of adenosine (10(-9) M to 10(-5) M). Muscles contracted in Krebs-Henseleit bicarbonate buffer (27 degrees C, 95% O2 and 5% CO2) for 500 ms at 50 Hz once every 90 s. Soleus fatigued significantly less with adenosine present at concentrations of 10(-8) M and higher than with the Krebs-Henseleit vehicle control. Adenosine significantly improved force generation or delayed fatigue of EDL only with the initial adenosine challenge. To investigate the receptor population involved, we exposed soleus to agonists specific for A1 receptors (N6-cyclopentyladenosine, CPA), or A2 receptors (CGS 21680 hydrochloride, CGS), or A3 receptors (N6-benzyl-5'-N-ethylcarboxamidoadenosine, BNECA). CPA (A1) significantly decreased fatigue compared with the Krebs-Henseleit vehicle control at concentrations of 10(-9) M and higher. Muscles exposed to the A2 and A3 agonists did not differ from a Krebs-Henseleit plus methanol control. Phenylephrine (10(-6) M), an alpha-adrenergic agonist that increases the concentration of inositol triphosphate (IP3), significantly improved developed force in soleus. Neither a permeable cAMP analog, 8-bromo-cAMP (10(-5) M), nor a beta, agonist, isoproterenol (10(-6) M), had an effect on force generation in the soleus when compared with a saline control. Thus adenosine slowed fatigue in slow-twitch skeletal muscle through A1 receptors.

Contractile function in vitro of slow-twitch skeletal muscle from weanling mice subjected to wasting malnutrition.

Our hypothesis was that malnutrition sufficient to produce weight loss in weanling mice would decrease the ability of slow-twitch skeletal muscle to develop and maintain force. We isolated muscles from 3 groups (n = 5) of weanling C57BL/6J mice of both sexes (i) mice at 19 days of age serving as zero-time or baseline controls (CONT) (ii) mice fed for the next 14 days with a low-protein diet that produces features of incipient kwashiorkor (LPD) and (iii) mice fed for the next 14 days with a complete diet (NORM). Muscles were also obtained from 5 adult mice 7-9 months of age (MAT). We stimulated the soleus at 50 Hz for 500 ms at 0.6 tetanic contractions per min (tet x min(-1)), 6 tet x min(-1), and 30 tet x min(-1) in Krebs-Henseleit bicarbonate buffer at 27 degrees C gassed with 95% O2 and 5% CO2. The initial developed force (mN x mm(-2)) at 0.6 tet x min(-1) did not differ across groups (CONT 211.7 +/- 16.0, LPD 274.2 +/- 41.6, NORM 246.8 +/- 38.0, MAT 210.8 +/- 10.6). The fatigue rate (mN x mm(-2) x min(-1)) at 6 tet x min(-1) was significantly slower in muscles from CONT (0.6 +/- 0.3) and LPD (0.6 +/- 0.4) than in NORM (2.4 +/- 0.6) and MAT (2.3 +/- 0.2). At 30 tet x min(-1), the fatigue rate (mN x mm(-2) x min(-1)) did not differ across groups (CONT 2.4 +/- 0.5, LPD 2.7 +/- 0.5, NORM 2.5 +/- 0.4, MAT 2.0 +/- 0.2). After stimulation at 6 tet x min(-1) and 30 tet x min(-1), only muscles from CONT and LPD recovered to 100%. Because muscles from LPD mice developed equal force, fatigued less, and recovered from fatigue to a greater extent than muscles from NORM mice, we rejected the hypothesis. The function of the tissue remaining in the muscles from LPD mice approximated that of muscles from mice at 19 days of age rather than muscles from either mice of the same age fed a complete diet or adult mice.

EDHF mediates the relaxation of stretched canine femoral arteries to acetylcholine.

To test the hypothesis that mechanically stretched arteries relax to endothelium-derived vasodilators, we challenged endothelium-intact dog femoral artery rings stretched from 1 to 16 g total initial tension (active force and passive elastic) with 10(-6) M acetylcholine (ACh), an endothelium-dependent dilator. The relaxation to 10(-6) M sodium nitroprusside (SNP), an endothelium-independent dilator, increased with the total initial tension. The relaxation to ACh averaged approximately 65% of the relaxation to SNP at total initial tensions of 4 to 16 g. To determine the nature of the endothelial-derived products involved, we compared the ACh-induced relaxation of stretched rings (6.5 +/- 0.2 g total initial tension) with rings chemically contracted with phenylephrine (Phe, 10(-7) to 10(-5) M) (6.5 +/- 0.3 g total initial tension). ACh-induced relaxation was evaluated before and after the inhibition of the synthesis of eicosanoids [cyclooxygenase (10(-5) M indomethacin) and lipoxygenase (10(-5) M nordihydroguariaretic acid)] and nitric oxide [nitric oxide synthase (10(-5) M Nw-nitro-L-arginine)]. The contribution of endothelium-derived hyperpolarizing factor (EDHF) was identified by blocking calcium-activated potassium channels (10(-8) M iberiotoxin). SNP (10(-6) M) relaxed stretched rings by 1.7 +/- 0.1 g and chemically-activated rings by 4.8 +/- 0.2 g. ACh relaxed stretched rings to 73 +/- 3% of the SNP relaxation and this was only attenuated in the presence of iberiotoxin. ACh relaxed Phe-activated rings to 60 +/- 3% of the SNP relaxation. This relaxation was attenuated by inhibition of the synthesis of nitric oxide and (or) eicosanoids. Therefore, ACh relaxed stretched rings through the release of EDHF whereas the relaxation of chemically activated rings to ACh involved multiple endothelium-derived vasodilators.

Polycythemia decreases fatigue in tetanic contractions of canine skeletal muscle.

PURPOSE: The effects of an acute polycythemia on muscle fatigue development were investigated in the self-perfused canine gastrocnemius in situ. METHODS: Following isolation of the gastrocnemius, dogs (N = 5) were made polycythemic through a bolus injection of packed erythrocytes (hematocrit (Hct) = 90-92%) to raise systemic Hct to 63.5 +/- 0.5%. Subsequently, the gastrocnemius was stimulated, through the sciatic nerve, to perform 20 min of isotonic tetanic contractions (60 x min(-1), 200 ms, 50Hz). Control (normocythemic) animals (N = 5) underwent an identical contraction regimen. RESULTS: Although blood flow to the gastrocnemius was not different at any time, oxygen delivery was significantly increased during polycythemia (peak = 33.7 +/- 2.2 mL x 100 g(-1) x min-1) over control (peak = 25.1 +/- 2.1 mL x 100 g(-1)x min(-1)) at all times during contraction. Oxygen uptake by the gastrocnemius, although consistently increased, was not significantly different between the normocythemic and polycythemic conditions at any time. The rate of fatigue was significantly decreased over the first 6 min of contraction in polycythemic animals (3.5 +/- 0.6% x min(-1)) when compared with controls (5.8 +/- 0.7% x min(-1)). Subsequent fatigue development was not different between groups. As a result of the early rate differences in fatigue, however, the work production in polycythemic animals was significantly greater than in normocythemic dogs for the duration of the contraction period. CONCLUSION: We conclude that during high metabolic rate isotonic tetanic contractions, muscle fatigue development is diminished by polycythemia, but the ergogenic effect appears to be transient.

Extravascular adenosine influences endothelium-derived nitric oxide release from perfused dog semitendinosus artery.

We tested the hypothesis that extravascular adenosine induces the release of vasodilatory products from endothelial cells lining skeletal muscle vessels. Endothelium-intact (n = 35) and -denuded (n = 5) dog semitendinosus intramuscular arteries were isolated, cannulated, and placed in 100-mL baths containing Krebs-Henseleit bicarbonate buffer (Krebs) at 37 degrees C and gassed with 95% O2--5% CO2. Each vessel, as well as a parallel tubing segment (avascular control), was perfused at 3.5 +/- 0.2 mL/min (inflow pressure 94 +/- 2 mmHg; 1 mmHg = 133.3 Pa) with Krebs containing 100 microM phenylephrine, 6% dextran, and 15 units/mL superoxide dismutase. Perfusate from all segments dripped onto endothelium-denuded dog femoral artery rings. The addition of 10 microM acetylcholine to the perfusate to test the functional integrity of endothelium-intact donor segments did not alter resistance in vessel segments or change force in rings. The addition of 100 microM adenosine to the extravascular bath decreased resistance 1.5 +/- 0.4 mmHg.mL-1.min-1 in vessel segments but was without effect on downstream rings. When acetylcholine was retested in the presence of extravascular adenosine, a relaxation (16 +/- 6%) occurred in rings receiving perfusate from endothelium-intact segments but not endothelium-denuded or tubing segments. This relaxation was eliminated by N omega-nitro-L-arginine (10 microM), a nitric oxide synthase inhibitor, and was attenuated to 4 +/- 1% by 8-phenyltheophylline (10 microM), an adenosine receptor antagonist. Thus adenosine, in conjunction with acetylcholine, acting through a receptor-mediated event, resulted in the release of nitric oxide from the endothelium of perfused intramuscular arteries, indicating the potential for extravascular conditions to influence the release of endothelium-derived products.

Microvascular hematocrit and permeability-surface area product in contracting canine skeletal muscle in situ.

The dynamics of the microvasculature of the blood-perfused canine gastrocnemius-plantaris muscle in situ at rest and during contraction were determined using multiple-indicator dilution analysis. Permeability-surface area product (PS) was estimated using a bolus indicator dilution of 86Rb, with 125I-albumin serving as the reference tracer, while microvascular hematocrit (Hmv) was estimated using the relationship between plasma (125I-albumin) and erythrocyte (51Cr) tracer washout curves. The muscle was stimulated to contract, under self-perfusion, for 3-min periods with either isometric twitch (1.5, 3, or 5 Hz; 4 ms) or tetanic (20, 40, or 60 trains/min, 200 ms, 100 Hz) contractions, separated by 25 min of rest, randomized to prevent ordering effects. At all stimulation frequencies, Hmv increased significantly from rest value of 36.5 +/- 1.6% to 3 min of either isometric twitch or tetanic contractions. PS rose significantly from 0.08 +/- 0.002 ml/g min at rest to a maximum of 0. 40 +/- 0.01 ml/g min at 60 isometric tetanic contractions per minute and 0.38 +/- 0.01 at 5 Hz. Changes in PS appeared related to stimulation frequency in both twitch and tetanic contractions. The change in Hmv with muscle contraction appeared to depend on contraction frequency during twitch contractions only, but was independent of stimulus frequency during tetanic contractions.

The effect of nitric oxide and endothelin on skeletal muscle contractility changes when stimulation is altered.

To investigate the effect of endothelial-derived products on the force of contraction of blood-perfused skeletal muscle, we infused S-nitroso-N-acetylpenicillamine (SNAP, 10(-4) M) (a nitric oxide (NO) donor), endothelin-1 (ET-1, 10(-8) M), N-acetylpenicillamine (NAP, 10(-4) M), or saline at a constant vascular concentration into the vascular bed of pump-perfused dog gastrocnemius-plantaris muscles in situ (n = 17). Muscles performed isometric twitch contractions at 0.5, 1.5, and 4 Hz and isometric tetanic contractions (150 ms, 50 Hz) at 12 and 40 contractions/min. We perfused the muscle at a constant pressure at rest and for the first 3 min of the 6-min contraction period, and then switched to constant flow perfusion and infused the substance over the remaining 3 min. Neither NAP nor saline had a significant effect on force of contraction or perfusion pressure. SNAP significantly attenuated developed force as compared with NAP at 40 contractions/min and at 1.5 and 4 Hz. The effect of SNAP on developed force was greater during twitch than tetanic contractions. ET-1 had no significant effect on twitch or tetanic developed force. To test for these results on another mammalian skeletal muscle preparation, we stimulated curarized trimmed mouse soleus in vitro for 500 ms at 50 Hz at either 2 or 4 contractions/min in the presence of SNAP (10(-4) M) or ET-1 (10(-9) M). SNAP, which increased force at 1 contraction/90 s, had no significant effect at the higher contraction frequencies. The inhibition by ET-1 at 2 contractions/min disappeared at 4 contractions/min. Therefore the effect of both NO and ET-1 on mammalian skeletal muscle appears to be dependent upon contraction pattern and frequency.

Mechanisms within the spinal cord are involved in the movement-induced attenuation of an H reflex in the dog.

1. H reflexes were elicited in the second interosseous muscle of the hindpaw of the anesthetized dog during passive rotation of the shank about the ipsilateral or contralateral knee. Reflexes sampled at four points in the cycle of movement were compared with stationary controls. For both the ipsilateral and contralateral limb manipulations, reflexes were significantly reduced (P < 0.05) across the cycle of movement. Position-related modulation of the reflex amplitude was not detected (P > 0.05) in either instance. 2. The experiments were then repeated after the spinal transection of each animal at the level of T13. Passive rotation about either the ipsilateral or contralateral knee significantly attenuated (P < 0.05) the H reflex across a cycle of movement in the spinal dog. There was little difference in the amount of inhibition produced by the movement between the intact and spinal animals. On average, the reflex was attenuated 29 +/- 2.4% (mean +/- SE) in the intact animals and 32 +/- 2.1% in the spinal animals. 3. It is concluded that passive rotation about the knee of either leg leads to suppression of the H reflex of the second interosseous muscle both in the ipsilateral, moving leg and the contralateral, stationary one. This reflex suppression occurs across the cycle of movement. The mediating circuitry lies within the spinal cord, caudal to T13.

Cultured dog aortic endothelial cells release vasodilatory products in response to acetylcholine and adenosine.

We tested the hypothesis that products released from contracting skeletal muscle could induce the release of vasodilatory products from vascular endothelium. Superfused endothelium-denuded rabbit aortic and dog femoral artery rings contracted with 1 microM phenylephrine delivered at 1-2 mL/min were subjected to 3-mL bolus challenges that had been briefly exposed to either a culture dish devoid of endothelial cells (control) or a dish containing dog aortic endothelial cells (experimental) immediately prior to their application. Challenges were alternated and included Krebs-Henseleit bicarbonate buffer (Krebs) pH 7.4 (vehicle); 1 microM acetylcholine; and the muscle metabolites 6 mM potassium chloride, 1 mM sodium phosphate monobasic, 10 microM adenosine, acidic Krebs pH 6.8, and 100 microM ammonium chloride. Only the products released from cultured endothelial cells after the addition of acetylcholine and adenosine resulted in a significant relaxation of vascular rings compared with control. Prior incubation of the cultured endothelial cells with 10 microM 8-phenyltheophylline eliminated the relaxation induced by adenosine, and all relaxations were eliminated by prior incubation of the cells with either 10 microM N omega-nitro-L-arginine or a combination of 10 microM indomethacin and 10 microM nordihydroguaiaretic acid. This indicates that at least one metabolite released from contracting skeletal muscle could induce the release of vasodilatory products from the endothelium, which would act as a local amplifier of functional hyperemia.

Endothelial cell products alter mammalian skeletal muscle function in vitro.

We tested the hypothesis that endothelin and nitric oxide (NO) alter the force developed by fast-twitch and slow-twitch mammalian skeletal muscle, using a mouse skeletal muscle preparation trimmed to approximately 50% of the original diameter to decrease diffusion distances. We suspended trimmed soleus (SOL) and extensor digitorum longus (EDL) muscles in Krebs-Henseleit buffer (27 degrees C; pH 7.4) gassed with 95% O2 -5% CO2. Muscles were stimulated once every 90 s for 500 ms at 50 Hz for SOL and 100 Hz for EDL. The force developed by trimmed SOL was 223.8 +/- 9.1 mN/mm2 and by EDL was 247.3 +/- 9.4 mN/mm2. Endothelin 1 (ET-1) had no effect on EDL but significantly accelerated the rate of decrease of developed force of SOL at concentrations of 10(-10) mol/L and higher within 10 contractions. When ET-1 was removed, force returned toward control value. Endothelin 3 (ET-3) had no effect on either muscle. S-Nitroso-N-acetylpenicillamine (SNAP), a source of NO, increased developed force over time in both muscles, with a threshold of 10(-6) mol/L. The effect was evident within 5 contractions in both muscles. Force remained elevated above control values after the removal of SNAP. Thus ET-1 attenuated and NO amplified mammalian skeletal muscle function.

Inhibition of canine H reflexes during locomotor-like rotation about the knee arises from muscle mechanoreceptors in quadriceps.

1. H reflexes were elicited in the small muscles of the foot in the canine and human during passive locomotor-like rotation of the shank about the ipsilateral knee. The movement-induced effect was similar in the two species. In the anesthetized dog, the reflex gain was reduced by 36 +/- 8.4% (mean +/- SE) on average, compared with appropriate stationary controls. Reflexes in the human were reduced during movement to 45 +/- 3.5% of their stationary control values. 2. H reflexes were elicited in the anesthetized dog during passive locomotor-like rotation about the knee and were compared with reflexes obtained with the limb stationary. Populations of mechanoreceptors were then systematically removed to ascertain which group or groups provided the sensory input that leads to the decrease in reflex gain during movement. We hypothesized that the majority of the reflex attenuation could be attributed to muscle mechanoreceptors. 3. Reflexes continued to be significantly reduced (P < 0.05) during passive movement about the knee until the muscle mechanoreceptors of the quadriceps muscle group were deactivated. The removal of input from joint receptors or cutaneous receptors did not eliminate the gain reduction induced by the passive movement. 4. It is concluded that muscle mechanoreceptors of the quadriceps muscle group provide an inhibitory input to the H reflex pathway of the dog plantar muscle when the knee is passively moved in a locomotor-like fashion. This source of inhibition likely also contributes to the soleus H reflex gain reduction in humans.

Electrical stimulation and amino acid and ammonia metabolism in the canine gastrocnemius muscle.

This study examined the effects of electrical stimulation on amino acid and ammonia (NH3) metabolism in the isolated in situ canine gastrocnemius muscle preparation. Cut sciatic nerves of 10 mongrel dogs were stimulated at either 3 or 5 twitches/s (10 V, 0.2-ms duration) for 60 min. Muscle NH3 release dramatically increased on stimulation, and over 60 min the 3- and 5-Hz groups released 86.7 +/- 24.2 vs. 160.8 +/- 17.4 mumol.min-1.100 g-1 (P < 0.05) of NH3, respectively. Similarly, the intramuscular NH3 concentration was elevated (P < 0.05) above rest for both groups throughout stimulation, and it was higher (P < 0.05) at 5 min for the 5-Hz (82.7 +/- 2.4 mumol/100 g wet wt) than for the 3-Hz (67.4 +/- 7.4 mumol/100 g wet wt) group. Stimulation was also characterized by a large release of amino acids by both groups. The total amino acid release for 60 min was 415.4 +/- 64.9 vs. 193.3 +/- 56.2 (P < 0.05) mumol/100 g for the 3- and 5-Hz groups, respectively. However, there were no shifts or differences between groups in the intramuscular total amino acid pools. Glutamine (Gln) and alanine (Ala) dominated the amino acids released by muscle and together represented 35 and 46% of the total amino acids released over 60 min for the 3- and 5-Hz groups, respectively. The total release of Gln was higher (P < 0.05) for the 3-Hz (81.1 +/- 5.6 mumol/100 g) than for the 5-Hz (49.4 +/- 10.7 mumol/100 g) group, but there were no differences between groups in total Ala release. In contrast, both groups demonstrated an uptake of branched-chain amino acids (valine, isoleucine, and leucine) after 45 min of stimulation. These data show a stimulation-dependent production of NH3 and release of amino acid by the canine gastrocnemius muscle. These data further show that the degree of net muscle NH3 production is proportional to the frequency, whereas the degree of amino acid release is an inverse function of frequency.

Blood flow and pressure relationships which determine VO2max.

The role of O2 delivery in regulating VO2max has been studied in an isolated gastrocnemius-plantaris muscle preparation contracting in situ; recent data addressing this issue are presented. VO2 increases nonlinearly with stimulation frequency reaching a peak at 5 twitches.s-1 or 1 tet.s-1 (200 ms trains, 50 imp.s-1). Further increases in stimulation frequency result in a lower VO2. Measured VO2 maxima are less than predicted VO2 capacity, and peak VO2 during tetanic contractions is greater than that during twitches. Above 150 imp.min-1, VO2 is directly related to the level of blood flow attained as VO2/Q (arterial-venous O2 difference) is fixed by some unknown mechanism. Increasing blood flow, with a pump, during 1.s-1 tetanic contractions increases O2 diffusive conductance and peak VO2. When O2 delivery is reduced, ischemic hypoxia appears to result in more rapid reductions in muscle performance than hypoxic hypoxia because of decreases in perfusion pressure and Q. 31P-NMR studies reveal that reductions in creatine phosphate and energy charge are similar between ischemia and hypoxia suggesting a common regulator, O2. We conclude that VO2max is limited by O2 delivery as a result of a limited and uneven distribution of muscle blood flow. These limitations appear secondary to mechanical restraints imposed by contraction duty cycle and vascular compression.