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PubMed1984

Topographical localization of motor endplates in cryosections of whole human muscles.

The distribution of motor endplates in biceps brachii, tibialis anterior, and sartorius muscles from human adults was studied by staining longitudinal cryosections of whole muscle for cholinesterase. A special freezing technique was used to prevent the muscle from cracking before sectioning on a heavy cryostat microtome. The results from a large number of cryosections from biceps brachii and tibialis anterior muscles were analyzed by a computer and the topographical distributions of endplates in different views of the muscles were reconstructed. In the biceps brachii muscle, the endplates formed a fairly distinct, slightly V-shaped band through the middle of the two heads. In the tibialis anterior muscle, the majority of the endplates were superficially distributed along the whole muscle. In the longitudinal sections from the middle part of the muscle, they gave the pattern of a parabola with its apex at the proximal end of the muscle. In the sartorius muscle, the endplates were scattered throughout the muscle and no endplate band was observed. The findings are in accordance with results obtained 30 years ago in investigations of muscle from small children and stillborn infants.

Adult↗

PubMed2001

A strong constant magnetic field affects muscle tension development in bullfrog neuromuscular preparations.

Effects of a constant magnetic field (CMF) of 0.65 T on muscle tension over 9 h were studied in the neuromuscular preparation of the bullfrog sartorius muscle. Tension was developed every 30 min by stimulation of the sciatic nerve (nerve stimulation) or of the sartorius muscle itself (muscle stimulation). In sciatic nerve stimulation, tension decreased rapidly for the first 3-4 h at a similar rate in both test (exposed to CMF) and control muscles. However, the rate of decrease became smaller and almost leveled off after 3-4 h in the test muscles, whereas tension continued to decrease monotonically in control muscles. The slope of the decrease for these later periods was significantly different between the test and the control conditions. Accordingly, tension was larger in test than in control muscles. In muscle stimulation, tension decreased monotonically from the start of experiments in control muscles, while tension in test muscles maintained their initial values for almost 3 h. Thereafter they started to decrease with a similar rate to the control. Hence, tension was always larger in test than in control muscles. A similar pattern of temporal change was observed for the rate of rise of the maximum tension for nerve or muscle stimulation. However, a significant difference was detected only in the case of muscle stimulation. The present results showed that a strong CMF of 0.65 T had biological effects on tension development of the bullfrog sartorius muscle by stimulation of the sciatic nerve as well as by stimulation of muscle itself. The presence of a small AC magnetic field component leaves open the possibility of an AC, rather than a CMF effect.

Animals↗

PubMed1991

Diameter changes in arteriolar networks of contracting skeletal muscle.

The effect of muscular contraction on vessel diameter was studied in the arteriolar network of the exteriorized cat sartorius muscle during normal and elevated vascular tone. Dilation during 4 Hz motor-nerve stimulation was proportionately greatest in the third-order (transverse) arterioles and in vessels immediately upstream and downstream (P less than 0.01). This pattern of dilation was maintained with increased contraction frequency (30 Hz) and during concurrent sympathetic nerve stimulation (8 Hz). The pattern of constriction with sympathetic nerve stimulation alone showed a similar trend with the greatest response in the third-order and adjacent vessels. A model developed to estimate the resistance distribution in the arteriolar network, using data from earlier micropressure and vascular architecture studies in the sartorius muscle, allowed calculation of the resistance change during muscle contraction and sympathetic stimulation. Model predictions indicate that the third-order and adjacent vessels are the greatest site of resistance with both normal and elevated vascular tone. Thus these vessels were the site of greatest resistance change during muscle contraction. The more proximal, arcade vessels made lesser contributions to overall resistance changes, whereas the most distal, fifth-, and sixth-order arterioles appear not to be important in this regard. These findings indicate the third-order, transverse, arterioles are of special importance in regulating blood flow in the sartorius muscle.

Animals↗

PubMed1975

Tonic properties of the Flexor carpi radialis muscle of the male frog (Rana temporaria).

The mechanical acitvity of the isolated flexor carpi radialis muscle (FCRM) of the mature male frog (Rana temporaria) was compared to that of the sartorius muscle. Muscles were stimulated directly by massive shocks at 20 degrees C and isometric tension was recorded during single twitch, twitch series, long tetani and long tetanus series. Some groups of muscles were poisoned with IAA-N2 or FDNB. The FCRM showed a high resistance to fatigue during a tetanus series and a "fatigue contracture" during a twitch series. The rate of the tetanic tension rise was smaller by five to six times than in the sartorius muscle (0 to 50% Po) and the twitch/tetanus ratio was always very small. During continuous activity and after the blocking of recovery metabolic pathways, FCRM used its energetic stores two to three times slower than the sartorius muscle. These stores did not seem to be more important. The results indicate that the FCRM is not a "tonic" muscle but, quite possibly, is composed of several types of fibre like many other frog muscles. However, some characteristics of mechanical response cannot be explained by the common classification into two types of fibre.

Animals↗

PubMed2005

The influence of hypothermia on P2 receptor-mediated responses of frog skeletal muscle.

The contractile responses of isolated Rana ridibunda frog sartorius muscle contractions evoked by electrical field stimulation (EFS) were studied at three temperature conditions of 17, 22 and 27 degrees C. Temperature-dependent increase of muscle contractility was found. ATP (10-100 microM) concentration dependently inhibited the electrical field stimulation-evoked contractions of sartorius muscle at all three temperatures; this effect was significantly more prominent at a temperature of 17 degrees C than at other two temperatures. Adenosine (100 microM) also caused inhibition of electrical field stimulation-evoked contractions which was statistically identical at all three temperature conditions tested. A P2 receptor antagonist, pyridoxalphosphate-6-azophenyl-2',4'-disulphonic acid (PPADS, 10 microM) reduced the inhibitory effect of ATP at all three temperatures but did not affect inhibitory action of adenosine. In contrast, 8-(p-sulfophenyl)theophylline (8-SPT, 100 microM), a nonselective P1 receptor antagonist, abolished inhibitory effects of adenosine at all three temperature conditions but did not antagonize inhibition caused by ATP. In electrophysiological experiments, ATP (100 microM) and adenosine (100 microM) temperature dependently reduced end-plate currents recorded in sartorius neuromuscular junction by voltage-clamp technique. The inhibitory effects of both agonists were enhanced with the decrease of temperature. 8-SPT (100 microM) abolished the inhibitory effect of adenosine but not ATP on end-plate currents. Suramin (100 microM), a nonselective P2 receptor antagonist, inhibited the action of ATP but not adenosine, while PPADS (10 microM) had no influence on the effects of either ATP or adenosine. It is concluded from this study that the effectiveness of P2 receptor-mediated inhibition of frog skeletal muscle contraction in contrast to that of adenosine is dependent on the temperature conditions.

Action Potentials↗

PubMed1957

Calcium equilibrium in muscle.

1. A study of the calcium equilibrium in isolated frog muscle has been attempted. 2. When sartorius muscles were immersed in Ca(45) Ringer's solution, the surface phase took up the Ca(45) in about 1 minute; the extracellular water space and connective tissue in about 30 minutes; and the intracellular space in about 300 minutes. 3. The percentages of total calcium in the whole muscle immersed in Ringer's solution was as follows: 10 per cent in the surface phase; 12 per cent in the extracellular water space; 17 per cent in the dry connective tissue; 24 per cent in the intracellular space; and 37 per cent as non-exchangeable calcium. 4. The exchange constants of isolated frog sartorius muscle to calcium has been determined. The flux of intracellular calcium in the steady state was approximately 0.8 mM/(liter hr). 5. It appears that there is a calcium pump pushing calcium out of the cell against an electrochemical gradient of about 4 cal./mM of calcium. However, since the flux is low, the maximum energy required per hour to pump calcium out of the cell against this high gradient is only about 2 cal./kg. muscle or about 1 per cent of the resting energy.

Animals↗

PubMed1988

Differential projections of B and C sympathetic axons in peripheral nerves of the bullfrog.

Accumulating evidence indicates that electrophysiologically distinct subsets of sympathetic neurons selectively innervate different classes of targets. The organization of this system may therefore be reflected in the sympathetic fiber contents of peripheral nerves. To test this possibility, we have mapped the pathways followed by three groups of postganglionic sympathetic axons in the bullfrog by recording compound action potentials and by retrograde tracing with horseradish peroxidase (HRP). The axons that were studied arise from fast B, slow B, and C-type neurons in ganglia 9 and 10 at the lumbar end of the paravertebral sympathetic chain. They project to peripheral targets primarily by way of the sciatic nerve and can be distinguished by the velocities with which they conduct action potentials. Action potentials were recorded with suction electrodes from isolated preparations composed of paravertebral chain ganglia 7-10, the sciatic nerve, and branches of the sciatic nerve that supply striated muscles, skin, and the bladder. Preganglionic B fibers were selectively activated by stimulating the paravertebral chain rostral to ganglion 7, and preganglionic C fibers were selectively activated by stimulating spinal nerves 7 and 8 at points central to their rami communicantes. Compound action potentials recorded from the sciatic, peroneal, tibial, and sural nerves and from the primary trunk of the pelvic nerve were each found to contain three components produced, respectively, by fast B, slow B, and C-type sympathetic axons. Similarly, action potentials recorded from cutaneous branches of the sciatic tree were found to contain three sympathetic components. By contrast, when compound action potentials were recorded from branches of the sciatic tree that directly enter and innervate striated muscles and also the bladder, the sympathetic responses were found to arise solely from C-type axons. HRP was used to label the sympathetic neurons that project to the sartorius muscle and into the cutaneous lateral crural nerve. Retrograde transport of HRP from the sartorius muscle labeled 17 +/- 4 (mean +/- s.d.) sympathetic neurons and 27 +/- 3 spinal motoneurons while transport from the lateral crural nerve labeled 68 +/- 47 sympathetic neurons but no spinal neurons. The average somatic diameter of ganglion cells projecting to the sartorius muscle was significantly smaller than that of cells projecting to the lateral crural nerve. The electrophysiological results indicate that fast B and slow B sympathetic axons in the sciatic trunk and its primary branches project selectively into cutaneous nerves while sympathetic C axons project into all peripheral nerves.(ABSTRACT TRUNCATED AT 400 WORDS)

Action Potentials↗

PubMed1988

Determination of fiber types of chicken skeletal muscles based on reaction for actomyosin, calcium+2, magnesium+2-dependent adenosine triphosphatase.

Muscle fiber subtypes, determined with the actomyosin Ca+2,Mg+2-adenosine triphosphatase (ATPase) reaction in chicken anterior latissimus dorsi and posterior latissimus dorsi muscles, were demonstrated only after acid or alkaline preincubation followed by a 60-min enzyme incubation. In contrast, subtypes were demonstrated in the sartorius muscle either with or without preincubation. A single-step procedure was therefore possible with this muscle. The results were generally similar to those obtained previously with the mycosin Ca+2-ATPase procedure. Both methods revealed corresponding muscle fiber subtypes, with the exceptions noted below. The actomyosin Ca+2,Mg+2-ATPase procedure, following preincubation at pH 9.4 and 10.3, resulted in a similar reaction intensity in all fiber types. With the myosin Ca+2-ATPase procedure, the IRA (slow) type in anterior latissimus dorsi and sartorius muscles and the I (slow), IIR (fast oxidative-glycolytic), and IIW (fast glycolytic) types in posterior latissimus dorsi muscle had a higher reaction intensity following preincubation at pH 9.4 than at pH 10.3. Fiber Types IIR and IIW in sartorius muscle were easily distinguished with the actomyosin Ca+2,Mg+2-ATPase procedure.

Actomyosin↗

PubMed1991

Can infected prosthetic grafts be salvaged with rotational muscle flaps?

The conventional approach to prosthetic graft infection, including graft removal and extraanatomic reconstruction, conveys a substantial risk of limb loss and death and mandates more innovative solutions. From January 1985 to January 1989 eight rotational muscle flaps were performed on four men and three women with prosthetic graft infection who ranged in age from 42 to 79 years (mean, 67 years). The grafts involved included aortofemoral (three patients), femoropopliteal (two patients), femorofemoral (one patient), and subclavian-carotid-carotid (one patient) and were composed of Dacron (five grafts) or polytetrafluorethylene (two grafts). Infections were in the groin in six patients and in the neck in the other patient; all patients had anastomotic exposure. Clinical presentations included abscess/purulent drainage (four patients), anastomotic hemorrhage (two patients), and anastomotic false aneurysm (one patient) and was associated with fever and/or leukocytosis in all patients. Positive bacterial cultures were obtained from all patients. Rotational muscle flaps performed included rectus abdominis (five grafts), pectoralis major (one graft), gracilis (one graft), tensor fascia lata (one graft); in two patients, rotational muscle flaps were performed after failed local sartorius muscle transfer. No major complications of the RMF procedures were encountered. One patient died 4 months after the operation of complications of ischemic colitis/perforation. The other six patients were discharged with completely healed wounds. One patient developed recurrent infection 12 months after the rotational muscle flap procedure; five (83%) patients have been followed for 12 to 51 months (mean, 27 months) without evidence of recurrent infection. These preliminary results suggest that rotational muscle flaps are a safe and effective treatment for prosthetic graft infection, even when local sartorius muscle coverage has failed.

Adult↗

PubMed1978

Maintenance of low sodium and high potassium levels in resting muscle cells.

1. Previous work has shown that a frog sartorius muscle consists of parallel cells running all the way from one end of the muscle to the other and that amputation of one end of the muscle is not followed by regeneration of a new cell membrane. If now only the cut end of the amputated muscle is exposed to a Ringer solution in which the solutes 42K and 22Na act as radioactive labels and the rest of the cell is suspended in air, we have what is described as an effectively membraneless open-ended cell or EMOC preparation. In this case the only remaining anatomically intact plasma membrane and pumps are made nonfunctional by the removal of 'sources' for inward pumps and 'sinks' for outward pumps. 2. The healthy region of a frog sartorius muscle EMOC preparation continues to accumulate labelled K+ to a level higher than that in the Ringer solution and to exclude labelled Na+ to a level below that in the Ringer solution, much as a normal uncut muscle does in its normal environment. The differences were reduced by inclusion of ouabain in the medium. 3. The diffusion coefficient of Na+ in the normal muscle cytoplasm at 25 degrees C was measured using two methods. The average diffusion coefficient measured was 2.07 X 10(-6) cm2/sec, roughly 1/6 that of the diffusion coefficient of Na+ in a 0.1 N-NaCl solution. 4. The data obtained are discussed in terms of the association-induction hypothesis. In this theory asymmetrical solute distribution, basically an expression of a non-energy consuming metastable equilibrium state, is the result of specific combinations of two opposing mechanisms: adsorption which raises the level of the intracellular solute; and exclusion from cell water which tends to lower it.

Animals↗

PubMed1991

Investigations on the biochemical characteristics of chronically underperfused muscle.

During operation, biopsies from the gastrocnemius muscle and, in some cases, from the sartorius muscle were taken from 32 patients with peripheral arterial occlusive disease and from 5 subjects with normal peripheral circulation. In patients with inadequate circulation only during exercise, when compared with the control group, increased activities of enzymes involved in oxidative metabolism (malate dehydrogenase, nicotinamide adenine dinucleotide phosphate-dependent isocitrate dehydrogenase, cytochrome C oxidase, creatine kinase), in amino acid metabolism (asparate aminotransferase, alanine aminotransferase), and in anaerobic glycoysis (lactate dehydrogenase) were found. In patients with circulatory disturbances that manifested themselves already at rest, enzyme activities were, with the exception of LDH, lower than those of patients with exclusively exercise-related insufficiency. By means of intraindividual comparisons with the corresponding enzyme activities in the sartorius muscle, the author was able to show that the changes found were not simply the result of differences in training conditions. The diminished concentrations of energy-rich phosphate are an expression of the anaerobic metabolic state in patients with inadequate circulation at rest. It is concluded that chronic ischemia of muscle leads to changes in the energy metabolism of the cell. In the presence of more nearly adequate circulation at rest, the portion of oxidative potential of the total energy metabolism increases. In contrast, if there is an inadequate circulation at rest, the mainly anaerobic glycolysis becomes quantitatively predominant.

Amino Acids↗

PubMed1995

Pharmacological characterization of the rhythmic synaptic drive onto lumbosacral motoneurons in the chick embryo spinal cord.

The isolated spinal cord of the chick embryo generates episodes of rhythmic bursting in which sartorius (hip flexor) and femorotibialis (knee extensor) motoneurons exhibit characteristic patterns of activity. At the beginning of each cycle both sets of motoneurons discharge synchronously. Following this brief synchronous activation sartorius motoneurons stop firing at the time of peak femorotibialis activity, producing a period of alternation between the two sets of motoneurons. Intracellular recording from motoneurons has suggested that the pause is mediated by a synaptically induced shunt conductance. However, the pharmacological basis for this shunt and the nature of the excitatory drive to motoneurons is unknown. To address these questions we have investigated the pharmacology of the rhythmic, synaptic drive to lumbosacral motoneurons using local and bath application of several excitatory and inhibitory antagonists, and documenting their effects on motor output in E10-E12 chick embryos. Local application of bicuculline or picrotoxin over sartorius motoneurons abolished the pause in firing recorded from the sartorius muscle nerve. As a consequence, the pattern of sartorius and femorotibialis activity was similar and the motoneurons were coactive. The pause in sartorius firing was shortened following local application of the glycine antagonist strychnine the nicotinic, cholinergic antagonists mecamylamine, and dihydro-beta-erythroidine and several excitatory amino acid antagonists. Application of the GABA uptake inhibitor nipecotic acid depressed the slow potentials and discharge recorded from the sartorius muscle nerve. These findings suggest that the pause is determined primarily by synaptic inputs acting at motoneuron GABAA receptors with contributions from glycinergic, cholinergic, and glutamatergic inputs. The actions of locally applied GABA onto spinal neurons are consistent with these findings because the neurotransmitter depolarizes spinal neurons and reduces their input resistance. Local application of bicuculline, but not strychnine, onto segments containing femorotibialis motoneurons altered the amplitude and duration of femorotibialis discharge and changed the profile of the slow potentials recorded from the muscle nerve. This finding implicates GABAergic inputs in the regulation of femorotibialis discharge. The pause in sartorius firing was still present and a pause in firing appeared in each cycle of femorotibialis discharge following bath application of bicuculline or strychnine. The pause in both sets of motoneurons could be abolished by local application of the NMDA receptor antagonist AP-5 onto the motoneurons, but not by local application of bicuculline. This action of AP-5 was in contrast to its activity in normal Tyrode's solution where it shortened the pause slightly.(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗

PubMed1986

Is the change in intracellular pH during fatigue large enough to be the main cause of fatigue?

The intracellular pH of frog sartorius muscles exposed to an extracellular pH 8.0 (25 mM HCO3-, 1% CO2) was 6.9-7.1. Following a fatiguing stimulation period (one tetanic contraction per second for 3 min), the intracellular pH was 6.5-6.7. When similar experiments were repeated with frog sartorius muscles exposed to pH 6.4 (2mM HCO3-, 1% CO2), the intracellular pH was 6.8-6.9 at rest and 6.3-6.4 following fatigue. So, in both experiments the intracellular pH decreased by 0.4-0.5 pH unit during fatigue. When the CO2 concentration of the bathing solution was increased from 1 to 30%, the intracellular pH of resting muscles decreased from 7.0 to 6.2-6.3. Although the effect of CO2 on the intracellular pH was greater than the fatigue effect, the decrease in tetanic force with CO2 was less than 40%, while during fatigue the tetanic force decreased by at least 70%. Therefore in frog sartorius muscle the decrease in tetanic force during fatigue exceeds the decrease that is expected from just a change in intracellular pH.

Animals↗

PubMed1978

An in-vitro model of malignant hyperthermia: differential effects of inhalation anesthetics on caffeine-induced muscle contractures.

Clinical concentrations of anesthetics augment caffeine-induced contracture of frog sartorius muscle; however, anesthetics differ in this characteristic. The potentiation was quantitated using six paired sartorius muscles for each specified concentration of anesthetic and controls. At a concentration of 1 MAC, the greatest potentiation occurred with 2 mM caffeine for all anesthetics studied. Under these conditions the order of magnitude of augmentations was: chloroform (15 times); halothane (11 times); methoxyflurane (10 times); cyclopropane (5 times); enflurane (4 times); isoflurane (3 times); diethyl ether (2 times); Baxter 3224 (2 times); fluroxene (1.4 times); nitrous oxide (1.3 times). Halothane at .5 MAC augments the 2 mM caffeine-induced contracture almost seven times, and at 2 MAC almost 13 times, whereas 2 MAC isoflurane potentiates the caffeine-induced contracture only four times and 4 MAC diethyl ether only two and a half times. It is postulated that those anesthetics that most potentiate caffeine-induced contracture may be the most potent triggering agents of malignant hyperthermia.

Anesthetics↗

PubMed1984

Distribution of capillary blood flow in the microcirculation of the hamster: an in vivo study using epifluorescent microscopy.

In vivo epifluorescent microscopy (EPI) was used to study capillary perfusion in superfused hamster cheek pouch, and cremaster and sartorius muscle preparations. In cheek pouches and cremaster muscles, in vivo epifluorescence microscopy (EPI) was compared to in vivo transillumination microscopy (TRANS) and the former was found to allow detection of a larger number of capillaries--34% more capillaries in cheek pouch and 21% more in cremasters were observed with EPI. The fraction of capillaries containing erythrocytes alone, plasma alone or plasma plus red cells was determined in all three tissues. Also, the fraction of capillaries unperfused was noted. Less than 2% of the capillaries contained plasma alone. The number of capillaries which contained stationary erythrocytes varied with vasomotor tone. In control cremaster and sartorius muscles 17 and 13% of observed capillaries were unperfused but contained erythrocytes. Ninety-eight percent of capillaries contained stationary erythrocytes in cremaster muscles vasoconstricted with 21% oxygen. From these observations we conclude (1) functional capillary counts obtained with TRANS may represent underestimates of the true number; (2) plasma shunts do not appear to be a significant factor in normal microcirculatory function in the hamster cheek pouch, or in cremaster and sartorius muscles; (3) since unperfused capillaries contain red cells, red cell counts in histological sections will overestimate functional capillarity. The magnitude of the overestimation will be a function of vascular tone and capillary hematocrit.

Animals↗

PubMed1999

Response of chick lines selected on carcass quality to dietary lysine supply: live performance and muscle development.

Broiler carcass quality can be improved by conventional selection techniques. In this regard, an experimental "quality" line (QL) was selected for high breast meat yield. We analyzed the effects of this selection on the dietary lysine requirement in chicks from 0 to 3 wk. Control (CL) and QL chicks were provided ad libitum access to isoenergetic diets containing 20% crude protein but differing in their lysine content (0.75, 0.88, 1.01, and 1.13%). Two-way ANOVA showed a significant effect (P < 0.01) of genotype on body weight, growth rate, feed intake, and weight of Pectoralis major and Gastrocnemius muscles. Conversely, the Sartorius muscle weight was not modified (P = 0.21) by genotype. Lysine deficiency markedly reduced body weight, growth rate, and feed intake, and increased feed conversion ratio (P < 0.001). Low dietary levels of lysine also depressed the weight of Gastrocnemius, Sartorius, and P. major (P < 0.001). The body or muscle weight response to diet lysine concentration depended on the line, with QL chicks appearing less sensitive to lysine deficiency. Consequently, their dietary requirements could be lower. Finally, when weight gain and P. major muscle protein deposition were plotted against lysine intake, QL chicks appeared to be more efficient than CL chicks. The underlying mechanisms responsible for this await clarification.

Animal Nutritional Physiological Phenomena↗

PubMed1959

Calcium influx in skeletal muscle at rest, during activity, and during potassium contracture.

Calcium influx in the sartorius muscle of the frog (Rana pipiens) has been estimated from the rate of entry of Ca(45). In the unstimulated preparation it is about equal to what has been reported for squid giant axons, but that per impulse is at least 30 times greater than in nerve fibers. The enhanced twitch when NO(-) (2) replaces Cl(-) in Ringer's is associated with at least a 60 per cent increase in influx during activity, whereas this anion substitution does not affect the passive influx significantly. Calcium entry during potassium contracture is even more markedly augmented than during electrical stimulation, but only at the beginning of the contracture; thus, when a brief Ca(45) exposure precedes excess K(+) application, C(45) uptake is increased three- to fivefold over the controls not subjected to K(+), whereas when C(45) and K(+) are added together, no measurable increase in Ca(45) uptake occurs. These findings are in keeping with the brevity of potassium contracture in "fast (twitch)" fibers such as in sartorius muscle.

Calcium↗

PubMed1984

Acetylcholinesterase activity in intact and homogenized skeletal muscle of the frog.

Enzymatic hydrolysis of acetylcholine (ACh) was determined in intact frog sartorius muscles or their homogenates. The Vmax was 29 nmol min-1 in intact muscles and 46 nmol min-1 per muscle in homogenates, and the Km was 6 and 0.2 mM, respectively. The muscle was divided into small segments, which were homogenized; the junctional cholinesterase (ChE) accounted for 60% of total enzyme activity. At low substrate concentrations the rate of hydrolysis was up to 30 times higher in homogenates than in intact muscles. This difference was greatly reduced at very high substrate concentrations. It appears that most of the ChE in intact muscle is 'occluded' to external ACh, mainly because the ChE at the edges of the synaptic cleft prevents the ACh from reaching the enzyme situated further inwards, which consequently does not contribute to its hydrolysis; homogenization makes all synaptic ChE accessible to added ACh. Incubation of sartorius muscles with collagenase caused an 80% decrease in ChE activity (determined in homogenates) of end-plate-containing parts which became similar to that in end-plate-free parts on which collagenase had little effect. Histochemistry showed that the tendon-muscle junction contained folds which were stained intensively for ChE. Diethyldimethylpyrophosphonate , neostigmine, eserine, and di-isopropyl fluorophosphonate inhibited ChE activity in this order of potency. The I50 values (i.e. the concentrations of the drugs which caused a 50% inhibition) were about 5 times higher in intact than in homogenized tissue. Neostigmine, 0.15 and 0.4 microM, increased the time constant of miniature end-plate currents 1.3- and 1.8-fold, and slowed down ChE activity of muscle homogenates by 1.4 and 2.1 times, respectively, without significantly affecting ACh hydrolysis by intact muscles. This indicates that synaptic ChE is not present in large excess. It is concluded that ChE activity measured in homogenates presents a better picture of in situ ChE activity than that measured in whole muscles especially for evaluating the effect of ChE inhibitors. A mathematical model for ChE-hindered diffusion of ACh is presented in an Appendix.

Acetylcholine↗