Muscle spindle response to active muscle shortening in Bufo marinus.
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The distribution of muscle spindles (MS) in the extensor digitorum longus (EDL) muscle of the rat hind limb was morphologically studied on a series of longitudinal cryostat sections. The intrafusal muscle fibers were brownishly stained with the acetylcholinesterase reaction, and the mucopolysaccharide contained in the equatorial periaxial space was stained with alcian blue. This double-staining method made it easy for us to find the MS and to decide the equatorial portion and the whole extension of MS. From a series of camera lucida drawings the distribution of all MS was reconstructed on a sheet of paper and three-dimensionally imaged on a personal computer using image reconstructing software. The MS were distributed mainly in the superficial and lateral part of EDL muscle. Additionally, an ATPase reaction was employed to detect the red muscle fibers, and it was confirmed that their distribution of them is similar to that of MS.
In the limb muscles, the muscle spindles have been demonstrated to be important in the maintenance of static posture. This role is supported by the close proximity of the muscle spindles to motor units that develop small forces and are fatigue-resistant, and the greater effectiveness of the input from muscle spindle afferents onto the small motoneurons supplying these motor units. In masseter, input from the muscle spindles is more effective on the larger motoneurons. This suggests that the muscle spindles may be more important in masseter for the development of large, fast forces, rather than for the maintenance of static postures. Thus muscle spindles in masseter may be important in load compensation during chewing and for the development of powerful bite forces in aggressive or defensive situations.
Small samples of the masseter and temporalis muscles of six adult male cats were dissected free, snap-frozen and cryosectioned. Serial sections from areas with muscle spindles were histochemically stained for myofibrillar ATPase after incubation in acid and alkali buffer, and for reactivity to two different myosin isoform-specific antibodies. Differential reactivity to these four stains identified bag1, bag2 and chain intrafusal fibre types. The most typical distribution of fibres within the spindle was 1 bag1, 1 bag2 and 3-4 chains. Although a few spindles were found in the anterior area of the temporalis that attaches to the coronoid process of the mandible and in the anterior area of the superficial masseter, the vast majority were found in the deepest portion of the masseter. Here spindles were characteristically with a number of simple spindles fused together into complex units.
The jaw-closing muscles of two bovine fetuses average 1,177 muscle spindles on one side of the face: 549 in the masseter, 433 in the temporalis, 192 in the medial pterygoid, and three in the lateral pterygoid. The jaw-opening muscles have no spindles.
Muscle-spindle density and size, in relation to extrafusal fibre-type, were examined in different portions of the masseter of six normal males by enzyme histochemistry. A direct relationship between spindle density, intrafusal fibre density and spindle size, and proportion of type I muscle fibres was found in the deep portion. Out of 410 muscle spindles in random samplings, 74 per cent were in the deep portion which is predominantly composed of type I fibres. Spindle density was three times greater than in the other portions, and spindles were commonly clustered, sometimes sharing capsule tissue. The deep masseter spindles were distinctive; they had the largest diameter and the most intrafusal fibres, the fibre density being four times greater than in other portions. Eleven per cent of the spindles, mainly in the deep portion, contained an unusually high number of intrafusal fibres (15 or more fibres). The findings imply that individual portions of the masseter have specialized functions. An especially powerful proprioceptive reflex mechanism is suggested for the deep portion. Thus, the human masseter may be part of an intricate, evolutionarily advanced, motor system also connected with speech.
Muscle biopsies were obtained from the erector spinae to investigate ultrastructural changes in nerve fibers and muscle spindles in patients suffering from adolescent idiopathic scoliosis. Membranous bodies are found in myelinated nerve fibers, both within and outside the muscle spindles. Leptofibrils are formed beneath the sarcolemma of the intrafusal muscle fibers. Additionally, in the intrafusal muscle fiber, there are slight increases in the amount of lipid droplets and glycogen particles, and the sarcoplasmic reticulum is slightly dilated. These changes may affect the metabolism of the muscle fiber and the normal balance of the contractile system. The cause-and-effect relationship of the changes in adolescent scoliosis warrants further investigation.
Muscle spindles from lumbricalis muscles of the rat were incubated for acetylcholinesterase with a modified thiocholine-method of Lewis and Shute and examined by light ane electron microscopy. All types of motor nerve ending showed heavy deposits of reaction product in the synaptic cleft. The underlying sarcoplasmic reticulum, transverse tubular system, and, when present, the envelope of sole plate nuclei were also stained. In the sensory region, the reaction was negative in the interface between the plasma membranes of the primary sensory terminal and muscle. One of two secondary sensory endings identified showed distinct reaction product in the cleft; the other secondary sensory ending showed no such reaction. Precipitates were present on the sarcolemma of the intrafusal muscle fibers in the polar and adjacent myotube regions, but not at the spindle equator. Extrafusal and intrafusal myelinated gamma-nerve fibers and preterminal motor axons showed staining of the axolemma. Fibers with thick myelin sheaths and preterminal sensory axons were free of acetylcholinesterase activity, as were the unmyelinated nerve fibers.
The morphological and histochemical alterations in the muscle spindles and Golgi tendon organs of tenotomized or immobilized calf muscles of rats (m. soleus, m. gastrocnemius) were studied in 54 animals. The intact contralateral feet served as controls. There was no change in the number of mechanoreceptors in either the tenotomized or the immobilized muscles. The diameter of the intrafusal fibers was increased somewhat after 1 week, but decreased by 14% to 40% in 3 weeks. The periaxial space of muscle spindle had diminished or disappeared by 2 weeks from the beginning of the procedures. The thickness of the outer capsule of the spindles had markedly increased, especially in the polar region. In the Golgi tendon organs the capsule had also thickened significantly and the internal space diminished. The alterations were similar following tenotomy or immobilization, but more marked after tenotomy than after immobilization.
In the present study muscle spindles (MSps) and Golgi tendon organs (GTOs) in bovine extraocular muscles (EOMs) were analyzed in detail. The innervation pattern of these proprioceptors was investigated with transmission electron microscope and confocal laser scanning microscope after double-fluorescent labeling. Three-dimensional (3D) reconstructions were performed of GTOs. Muscle spindles. MSps are numerous, each containing two nuclear bag fibers and up to eight nuclear chain fibers. In the equatorial region and paraequatorial region thin axons enwrapping the intrafusal muscle fibers form numerous nerve contacts on the muscle fiber surface. Double staining of such nerve terminals with synaptophysin and alpha-bungarotoxin and their fine structural features confirm their sensory nature. In the encapsulated part of the polar region neuromuscular contacts have structural features of motor nerve terminals and stain positively with alpha-bungarotoxin. Golgi tendon organs. GTOs are numerous in bovine EOMs. Each GTO contains collagen bundles but frequently also intracapsular muscle fibers. Intracapsular muscle fibers either terminate inside the GTO in collagen bundles or pass through the proprioceptor. GTOs are richly supplied with sensory nerve terminals which intermingle with the collagen bundles. Nerve terminals on intracapsular muscle fibers exhibit fine structural characteristics of motor nerve terminals and are alpha-bungarotoxin positive. The 3D images of GTOs show the detailed spatial arrangement of the GTO tissue components. These new insights in the complex and specific morphology of MSps and GTOs in bovine EOMs indicate that we deal with highly developed proprioceptors. These are supposed to provide important information for EOM innervation.
1. Muscle spindle afferent discharges exhibiting an approximately linear length-frequency relation could be recorded from the phrenic nerve in the isolated phrenic nerve-diaphragm preparation of the rat. 2. Muscle spindle afferent discharges could be identified by their characteristic "spindle pause" during muscle contraction and by their response to succinylcholine. 3. Cholinergic influence on spontaneous and stretch-induced afferent discharges was indicated by the augmentation produced by physostigmine and acetylcholine. (+)-Tubocurarine, but not atropine, prevented this augmentation indicating the presence of curariform cholinoceptors in muscle spindles. 4. Acetylcholine did not appear to be involved in the genesis of spindle afferent discharges as incubation with hemicholinium-3 and (+)-tubocurarine failed to affect the rate of spontaneous and stretch-induced spindle discharges. 5. Oxotremorine markedly increased the rate of spontaneous and stretch-induced spindle afferent discharges and this effect was prevented in the presence of hemicholinium-3 and (+)-tubocurarine. 6. These results with oxotremorine are of interest in connection with the observation that muscle spindle afferents and hyperactive in Parkinsonian patients.
In decapsulated muscle spindles, application of dibutyryl cyclic AMP (d-cAMP, 0.1-10 mM) and forskolin (10-100 microM) increased the rate of spontaneous discharges and decreased the responsiveness to stretch. Addition of 2-5 mM CaCl2 or 0.5-2 mM BaCl2 to the above drugs prevented the deterioration of the responsiveness to stretch and the increase in the rate of spontaneous discharges. Similar changes in the afferent discharges were observed with application of isobutylmethyl-xanthine (IBMX, 0.5-2 mM) or carbonyl cyanide chlorophenylhydrazone (CCCP, 1-10 microM). Application of 0.1-1 mM quercetin resulted in a prolonged increase in the discharge rate, lasting 15-20 s after the release of stretch. These results suggest that cAMP is involved in the regulation of the sensory processes through a modification of intracellular calcium activity.
The regularity of receptor discharges of passive muscle spindles adapted to a different length of the triceps of the calf of narcotized cats was investigated in the range of average frequencies from 5.6 to 40 impulses/sec. The values of the coefficients of variation of the impulse intervals in discharges of the primary endings are distributed in the range from 2.5 to 18.0% and of the secondary from 1.2 to 12%. The discharges of the primary endings are less variable than the discharges of the primary. Among the latter are found receptors with less, moderately, and more regular discharges at any length of the muscle. The shape of the "average impulse interval vs. coefficient of variation" relationship is nonlinear and differs for different units. For samples of receptors having approximately the same variability of the impulse intervals the average values of the coefficients of variation decrease significantly with muscle tension and increase of the rate of impulse flow. In some cases a decrease of the values of the coefficients of variation occurs due to a decrease of the dispersion at the same rate of flow and in other cases both parameters change simultaneously. A decrease of the dispersion of the intervals in receptor discharges apparently reflects an increase of the number of uniformly depolarized endings as their tension increases.
1. The effects of the muscle-depolarizing drug succinylcholine (SCh) on the stretch responses of jaw-closer muscle spindle afferents were studied in the anaesthetized cat. Using ramp and hold stretches repeated every 6 s the basic measurements made were: initial frequency (IF), peak frequency (PF) and static index (SI), the frequency 0.5 s after the end of the ramp of stretch. Derived from these were: dynamic difference (DD) = PF-IF, dynamic index (DI) = PF-SI and static difference (SD) = SI-IF. Increases in these measures caused by a single I.V. dose of SCh (200 micrograms kg-1) are symbolized by the prefix delta. 2. In a population of 234 units, delta DD and delta IF were each distributed bimodally, but were uncorrelated, thus defining four subgroups. 3. delta DD was argued to be an index of the effect of bag1 intrafusal fibre contraction and delta IF to be an index of the effect of bag2 fibre contraction. On this basis it is proposed that units can be divided into four groups according to the predominant influences of the bag1, bag2 and chain fibres as b1c (6.8%), b1b2c (22.2%), b2c (54.3%) or c (16.7%). 4. Testing with sine wave stretches at 1 Hz showed that changes in mean frequency and amplitude of response caused by SCh correlated with delta IF and delta DD respectively, but separated groups of units much less effectively than did ramp and hold testing. 5. Evidence is presented to indicate that the control value of DD in passive spindles does not relate to the potential strength of bag1 fibre effects in fully activated spindles. The bag1 fibre appears to contribute little to responses of spindle afferents in the passive state. DD is superior to DI as a measure of bag1 effects. 6. Conduction velocity was unimodally distributed in masseter spindle afferents and was not correlated with delta DD or delta IF and was therefore of no value in classifying them. 7. Neither the threshold of afferents to quick transient stretch nor the coefficient of variation of interspike intervals provided any significant additional help in classification. 8. The unexpectedly high proportion of units of b2c type is thought to include primaries lacking appreciable bag1 fibre contacts and secondaries with more or less substantial bag2 contracts.
A model of the muscle spindle was developed based on its anatomical structure. The model contains three intrafusal fibers (bag1, bag2, and chain), two efferents (dynamic gamma efferent to the bag1 fiber and static gamma efferent to bag2 and chain fibers), and two afferents [primary (Ia) and secondary (II)]. As in the real muscle spindle, the spindle model, under the modulation of gamma efferents, responds to the extrafusal muscle fiber length. Both outputs (Ia and II afferents) of the model were compared extensively with published data, under both sinusoidal stretch (with different stretch amplitudes and frequencies) and ramp and hold stretch (with different stretch amplitudes and velocities) in three different fusimotor activation conditions (dynamic gamma stimulation, static gamma stimulation, and without gamma stimulation). Model Ia afferent responses fit the published data well with active gamma input, but less well in the passive state. Model II afferent responses also fit the published data, although less quantitative data were available for comparison. The model correctly predicted the fractional power dependence of the primary and secondary ending responses on stretch velocity. The current model provides a powerful tool for simulation studies of neuromusculoskeletal systems, and demonstrates the feasibility of using a structural approach to model complex neurophysiological systems.
The impulse activity of muscle spindles isolated from the cat tenuissimus muscle was investigated under varying concentrations of the L-type calcium channel blocker nifedipine. At a concentration of 25 microM nifedipine impulse activity was clearly diminished in both primary and secondary endings. However, low concentrations of the drug (5-10 microM) exerted unexpected excitatory effects. The dynamic properties of primary endings in particular were augmented; those of secondary endings were also increased, although only slightly. A detailed analysis of the afferent discharge patterns obtained under ramp-and-hold stretches yielded the following effects of 10 microM nifedipine. (1) The initial burst at the beginning of the ramp phase of a stretch was increased in primary endings; (2) the peak dynamic discharge frequency at the end of the ramp phase was considerably increased in most primary endings; (3) the sensitivity of the peak dynamic discharge value to varying amplitudes and velocities of stretch was significantly enhanced in primary endings, and also increased, although only slightly, in secondary endings; (4) the rise in the discharge frequency during the ramp phase of a stretch was augmented in both types of ending, the effect being again stronger in primary endings; (5) the fast adaptive decay of the impulse frequency following the ramp phase of a ramp-and-hold stretch was significantly increased in primary endings, but remained unaffected in secondary endings. The enhanced dynamic properties of primary endings were also observed under small sinusoidal stretch stimuli (10 microm, 40 Hz), where nifedipine induced a significant shift in the position of the 1:1 driven action potentials toward smaller phase values. In view of an increase in tension in the isolated muscle spindle and an increased initial burst in primary endings in the presence of nifedipine, it is suggested that the drug facilitates the attachment of cross-bridges in the poles of the intrafusal muscle fibers. The increase in the dynamic properties of primary endings points to the possibility that the drug preferentially affects the nuclear bag(1) fiber. The inhibitory effect on the afferent discharge rate at high doses of the drug is interpreted as the consequence of a calcium channel block in the membranes of the sensory endings. The membrane potential of sensory endings appears to be highly dependent on sustained Ca(2+) conductance.