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Morphometry and histoenzymology of the hamster tenuissimus and its muscle spindles.

Muscle spindles and extrafusal fibers in the tenuissimus muscle of mature golden Syrian hamsters were studied morphologically and quantitatively using several light microscopic techniques. Muscle spindles were identified in serial-transverse frozen-sections of whole muscles stained with hematoxylin and eosin. Five tenuissimus muscles were examined from origin to insertion, and the locations of individual receptors were plotted in camera-lucida reconstructions. Spindles were found in proximity to the main neurovascular bundle in the central core of each muscle. A range of 16-20 receptors was noted per muscle. The mean muscle spindle index (the total number of spindles per gram of muscle weight) was 503 and the average spindle length was 7.5 mm. Oxidative enzyme and myosin adenosine-triphosphatase (ATPase) staining profiles were also evaluated in the intrafusal and extrafusal fibers in each muscle. Even numbers of type I and type IIA extrafusal fibers were distributed homogeneously throughout all muscle cross-sections. Histochemical staining patterns varied along the lengths of the three intrafusal fiber types. Nuclear chain fibers possessed staining properties similar to the type IIA extrafusal fibers and exhibited no regional variations. Bag1 fibers displayed staining variability, particularly when treated for myosin ATPase under acid preincubation conditions. Some spindles were isolated under darkfield illumination and then either treated with 7-nitrobenz-2-oxa-1,3-diazole (NBD)-phallacidin to detect filamentous actin by fluorescence microscopy, or prepared for conventional scanning electron microscopy (SEM). By fluorescence microscopy, a registered actin banding-pattern was observed in the sarcomeres of the intrafusal fibers, and variations in the intensity of banding were noted amongst different fibers. SEM revealed punctate sensory nerve endings that adhered intimately to the surfaces of underlying intrafusal fibers in the equatorial and juxtaequatorial regions. By transmission electron microscopy (TEM) these endings appeared crescent-shaped and were enveloped by external laminae. Each profile contained numerous mitochondria and cytoskeletal organelles. The high spindle density observed in this muscle suggests that the hamster tenuissimus may function in hindlimb proprioception.

Animals

Spaced serial section analysis of the avian muscle spindle.

Muscle spindle ultrastructure of the extensor digitorum communis of pigeons was studied using spaced serial sections. The intrafusal fibers extended well beyond the ensheathing capsule, after which they became disoriented and often fused with each other before terminating on the connective tissue of extrafusal fibers. Several extracapsular fibers contained macrofibrils which were about 0.1 micron in diameter and contained several dozen smaller (10 nm) subunits. Intrafusal fibers commonly formed close attachments with one another for short or extended (240 micron) lengths. A basal lamina was absent between regions of pairing, and a myosatellite cell lay at the border of the coupled region. Several fibers could be coupled together in a single cross section; fibers coupled together, separated, and either recoupled or became associated with other fibers along the length of a spindle. Profiles of sensory terminals and sensory satellite cells alternated to form a smooth-contoured surface over most of the fiber cross section in the equatorial region. The sensory terminals contained many mitochondria, lysosomes, and clear and dense core microvesicles. Both the terminals and sensory satellite cells formed desmosomelike junctions with the intrafusal fiber. A crescentic collagen sheath covered that portion of the fiber cross section containing the sensory terminal-satellite cell complex. Inner capsule cells surrounded the entire assembly in the equatorial region. The basal lamina thicknesses differed over the naked intrafusal fiber compared to that portion covered by the sensory terminal or sensory satellite cell. The thickness was more than doubled over the latter regions, indicating that the basal lamina over these areas was a product of the fused intrafusal fiber and sensory terminal and/or sensory satellite cell basal laminae. These are discussed in terms of intrafusal fiber degeneration and regeneration.

Animals

Ultrastructural observations on muscle spindles in extraocular muscles of pig.

Human and some mammals such as the sheep, goat and domestic and wild pigs have more or less muscle spindles in the extrinsic eye muscles, especially the domestic pigs having abundant muscle spindles (Matsuyama, 1987). The muscle spindles play a large role in maintaining the stable visual posture of the eyeballs. To define the morphological properties of the muscle spindles relative to the eye movement, the ultrastructure of the spindles was investigated in 6 extraocular muscles of the pigs by electron microscopy. The muscle spindles in the pig extraocular muscles consist of 4 to 5 intrafusal muscle fibers, one of which is nuclear bag fiber and 3 to 4 are nuclear chain fibers. The outer capsule is thin, composing of few layers, and the inner capsule ramifying to enwrap the individual fiber, accompanied by the medullated and unmedullated nerve fibers and blood capillaries. The nuclear bag fiber, 14 micron in diameter, is innervated by the atypical annulospiral sensory terminals and the chain fiber by the typical annulospiral terminal packed with mitochondria and microvesicles. The intrafusal fibers are innervated by the flower-spray sensory terminals anchoring deeply into the sarcoplasma, having abundant neurotubules and few mitochondria. The gamma motor end-plates have a relatively smooth synaptic cleft with a width of 70 nm and synaptic boutons containing few synaptic vesicles, sometimes, revealing a shallow fold of postsynaptic sarcolemma and abundant synaptic vesicles. The alpha motor end-plates reveal a relatively smooth synaptic cleft with a width of 80 nm, sometimes with a rough postsynaptic infolding, and boutons containing few synaptic vesicles and small-sized mitochondria. The satellite cells are innervated by the sensory terminals in various ways. The muscle spindles in the pig extraocular muscles are found to be much simpler in structure than those in the other antigravity muscles of the body. Their ultrastructure seems to reflect the morphological adaptation relative to the eyebal movement.

Animals

Morphometric comparison of capillaries in muscle spindles, nerve, and muscle.

Morphometric study was performed on transverse sections of microdissected muscle spindles from rabbit tenuissimus muscles. It showed a statistically significant difference in size and structure between the intrafusal capillaries and those of extrafusal muscle. The former are larger in diameter, circumference, and area due to a proportionate increase in the number of endothelial cells. Vesicles within spindle endothelial cells are fewer, mitochondrial counts are greater, but in proportion to the increased number of endothelial cells, intercellular junctions are tight and pericyte coverage is greater. The basement membrane around endothelial cells and pericytes is thicker and more often multilayered. The endoneurial capillaries of tenuissimus intramuscular nerve are morphologically identical to intrafusal capillaries. The similarity of the capillaries of spindles to those in brain suggests that a blood-nervous system barrier extends from brain into the peripheral nerves and muscle spindles.

Animals

Effects of the infraorbital nerve-transection on the formation of Ia fibers in muscle spindles of the masseter muscle in developing mice.

The effect of the transection of the infraorbital nerve on d 20 after birth on the formation of the annulospiral ending of Ia fibers of muscle spindles in the masseter muscle was investigated in developing mice. On the 20th d after the operation, there were significant differences between control and treated animals in the ratio of muscle spindles with non-coiled endings to all muscle spindles, and the ratios of muscle spindles with non-coiled endings in the masseter muscle on the operated and sham-operated sides were about equal to each other. On the 30th d after the operation, the ratio for the masseter muscle on the operated side was approximately twice that for the one on the sham-operated side. The ratio for the operated side was about equal to that for 15-d-old control mice, and about twice that for 20-d-old control animals. Thus, the present results indicate that disorder of sensory inputs following the transection of the infraorbital nerve leads to degeneration and/or inhibition of the formation of annulospiral endings in muscle spindles of the masseter muscle, and suggest that disorder of the masticatory mechanism may be caused by the degeneration and/or inhibition of the formation of the endings in many muscle spindles of the masseter muscle.

Animals

[Ultrastructure of the muscle spindles in the extraocular muscles of pigs].

To define the morphological properties of muscle spindles relative to the accurate control of eyeball movement, muscle spindles were investigated in 6 extraocular muscles of three white pigs by an electron microscope. The muscle spindles were composed mainly of a thin outer capsule, nuclear bag and chain fibers. The bag fibers seem to be slow fibers and the chain fibers to be fast fibers in the external fashion, in so far as the former has a greater myofibril density than the latter. The intrafusal muscle fibers were innervated by three categories of nerve terminals; annulospiral and flower spray sensory terminals and gamma motor terminals. The annulospiral terminal was coiled sparsely on the bag fiber and densely on the chain fiber. The cross terminals were found in the pairing muscle fibers. We frequently observed that the satellite cells were controlled by the sensory terminals. It was also frequently observed that the muscle spindles were composed of only one intrafusal bag muscle fiber. It was suggested that the extraocular muscle spindles of the pig remained a primordial profile in the structure.

Animals

[Decrease in the frequency of muscle spindle discharges during muscle stretching].

When studying responses of 500 muscle receptors to stretching of the m. triceps of the cat deafferented hindlimb, 23 cases revealed a transient decrease in the firing rate (DFR) of muscle spindles' receptors. The DFR occurs in the same receptor at different forms and rates of stretching approximately at the same area of stretching ("the DFR area"). When increasing the rate of stretching, the DFR effect is enhanced. The DFR effect seems to be due to a short-lasting unloading of the spindle area which contains the receptor terminals. The unhomogenous structure of the extrafusal muscle as well as various mechanical properties of the intrafusal muscle fibers can provide necessary conditions for the above effect.

Animals

Structure of the intraspinal projections of single, identified muscle spindle afferents from neck muscles of the cat.

The morphology and frequency of collaterals originating from single afferents supplying primary endings of muscle spindles in dorsal neck muscles have been examined using intra-axonal injections of HRP. Within the segment in which the afferent entered the spinal cord, one collateral was found for every 3.3 mm of stained axon. In contrast, afferents--one of more segments rostral to the segment in which they entered the spinal cord--had fewer collaterals: One collateral was found for every 6.3 mm of stained axon. The branching structure and terminal distribution of the collaterals were generally similar regardless of the muscle from which the afferent originated and the segment in which the collateral was found. Boutons were found in 2 zones: One of these was located in the intermediate zone, within and around the central cervical nucleus, and the other was found in laminae VIII and IX, including the motoneuron nuclei. The ventral termination zone of collaterals in the same segment as their parent axon entered the spinal cord was larger and had more boutons than the same projection of collaterals whose parent axon entered the spinal cord 1 or 2 segments caudal to the segment in which the collateral was found. These results indicate that afferents supplying primary endings of neck muscle spindles are more likely to contact neurons in the same segment in which the afferent enters the spinal cord than in more rostral segments. However, even within the same segment in which the afferent enters the spinal cord, the projection of neck muscle afferents to the ventral horn is less dense than the corresponding projection of hindlimb muscle spindle afferents in the lumbosacral spinal cord.

Animals

Prolonged degeneration of muscle spindles in the masseter muscle after treatment of developing mice with the local anesthetic lidocaine hydrochloride.

The effect of lidocaine-HCl on muscle spindles in the masseter muscle of developing mice was investigated. Repeated injections of mice with anesthetic in the short term decreased the diameters of primary endings, intrafusal muscle fibers and outer capsules in the equatorial regions of muscle spindles, and caused a drop in the succinic dehydrogenase activity in intrafusal muscle fibers of the muscle spindles. In addition, the diameters did not recover to the control value even after about 10 weeks following cessation of anesthetic treatment. Thus, the present results suggest that repeated use of lidocaine-HCl in developing animals may cause dysfunction of the skeletal muscles.

Animals

[Muscle spindles in denervated and reinnervated m. soleus of the rat. I. Changes in number, distribution and length of muscle spindles].

After a transient or permanent unilateral denervation of the soleus muscle of the rat the number, distribution and length of muscle spindles were determined. The results were compared with those of the contralateral innervated muscle and with the data received from investigating soleus muscles of normal uninjured rats. Denervation (with or without reinnervation) reduces the number of muscle spindles by nearly 50%. The typical uniform distribution of muscle spindles in the muscle remains almost unchanged. The remaining muscle spindles grow longer. The same findings are observed in the contralateral still innervated muscles, too. Consequently a comparison of the denervated (reinnervated) muscle exclusively with its contralateral muscle does not show different counts of muscle spindles. Therefore in such investigations the contralateral muscle is of dubious value as a mean of control. As regards number, distribution and length of muscle spindles there exist only small differences between the various strains of rats.

Animals

Measurements of muscle stiffness, the electromyogram and activity in single muscle spindles of human flexor muscles following conditioning by passive stretch or contraction.

In experiments on adult human subjects we examined the effect on passive mechanical properties of a muscle by conditioning it with either an isometric contraction or passive muscle extension. The test measurement was the amount of muscle displacement (stiffness) and the accompanying EMG in response to a brief torque pulse. Two muscles were tested, flexor digitorum profundus (FDP) and brachialis. In FDP the discharge of single muscle spindles was recorded as well. After muscle extension and return to the initial length, passive stiffness was less than after an isometric contraction. The changes in stiffness were accompanied by changes in pattern of EMG and in the responses of muscle spindles. It is suggested that in resting muscle there are stable cross bridges between actin and myosin filaments of muscle fibres which largely determine the passive stiffness. Muscle extension leads to detachment of these cross bridges which then re-form at the longer length. Return of the muscle to its starting length leads to development of slack in muscle fibres because, stiffened by the presence of the stable cross bridges, they are unable to shorten. Slack in muscle fibres lowers their measured stiffness. Muscle contraction, on the other hand, will result in any preexisting slack being taken up by the actively shortening muscle fibres, thereby raising muscle stiffness. Stiffness in intrafusal fibres is likely to follow a similar pattern to that in extrafusal fibres, leading to changes in stretch responsiveness of muscle spindles and consequently in the reflex EMG. It is concluded that the changes in stiffness and accompanying reflexes observed in this study are likely to be seen, at least under some conditions, in normal movements.

Adult

Interaction of recurrent inhibitory and muscle spindle afferent feedback during muscle fatigue.

Mammalian skeletal motor units have differing properties including their different susceptibility to fatigue. The question discussed in this paper is whether and to what extent proprioceptive feedback via muscle spindles can contribute to shape the firing patterns of motor units so as to minimize their loss of force during fatiguing contraction. The firing of a skeleto-motoneuron dispatches signals which are fed back to the same and homonymous as well as synergistic motoneuron. Two feedback pathways are of concern here: one via the related muscle unit and muscle spindle afferents (proprioceptive path), and one via recurrent motor axon collaterals and Renshaw cells (recurrent inhibitory path). It is suggested that the contraction of a motor unit or a small group of adjacent ones is signalled to the homonymous alpha-motoneurons via proprioceptive afferents, the signal being filtered and enhanced by spinal recurrent inhibition. This is effected by timed correlation of the signals which are propagated through the two feedback loops. The effects of the correlation can be strengthened by (i) topographical order of the feedback connections, (ii) heterosynaptic interaction, and (iii) tendencies towards synchronous discharge between motoneurons. These mechanisms render the possibility more likely that information about the unfused contractions of a muscle unit (or a small group of them), mediated via proprioceptive afferents, play a role in shaping the precise discharge pattern of the innervating motoneuron(s). These mechanisms could be used to optimize the force output of large fatiguing motor units during long activation, during which their activation rates normally decrease (adapt) over time. Our results show that during adapting motoneuron firing Renshaw cells and muscle spindle afferents may show discharge patterns which at least in part are in keeping with such an hypothesis.

Animals

Topographic studies relating distribution of Ia- and gamma-fibres in spinal cord and position of muscle spindles in cat tibialis anterior muscle.

The segmental distribution of 115 Ia- and 115 paired gamma-fibres of the tibialis anterior muscle was studied in anaesthetized cats. All Ia-fibres recorded were found in the lumbar segments L6 and L7, from caudal L6 to middle L7. The paired gamma-axons were also mainly found in these parts of the spinal cord, only 7 gamma-fibres were localized in caudal L7. A total of 70% of all fibres was found in L7. Of the fibres constituting 'muscle spindle units' of the tibialis anterior 92.2% enter the same segment (a 'muscle spindle unit' is here defined as a muscle spindle with its Ia-fibres and one gamma-fibre innervating it). More than that, 88% of the afferent and efferent fibres of muscle spindle units were found in the same part of the segment. For the first time, the position of the muscle spindles was related to the location of their Ia- and gamma-fibres in the spinal cord. In general, the muscle spindles located in the proximal muscle region project to the more cranial part of the spinal cord and the muscle spindles localized distally in the muscle project to the more caudal part of the spinal cord. The topographic pattern of the muscle spindle units is discussed with respect to the topographically arranged monosynaptic reflex loop.

Animals

The responses of muscle spindles in sheep extraocular muscles.

1. Responses from de-efferented muscle spindles in sheep extraocular muscles have been recorded in the Gasserian ganglion using glass microelectrodes. 2. The afferent fibre conduction velocities ranged from 32 to 106 m/sec and most were intermediate. The distribution was unimodal. 3. The afferent responses to ramp stretches and longitudinal vibration of the muscle could not be divided into two groups, but both the dynamic indices and vibration sensitivities of the afferents increased with their conduction velocities. 4. Injection of suxamethonium did not help to classify afferents with intermediate conduction velocity. 5. It is concluded that the afferents from sheep extraocular muscle spindles form a single population whose properties are correlated with their nerve fibre diameters.

Animals

Histochemical profile of muscle spindles of rat's sural muscles.

The histochemical profile of intrafusal fibres of muscle spindles and muscle spindle capsule were analysed in normal sural muscles of a rat. Weak activity of oxidative enzymes (NADH diaphorase, succinic dehydrogenase, lactate dehydrogenase) and creatine phosphokinase were found in bag1 and bag2 fibres, but the oxidative enzyme activity was moderate in chain fibres. High phosphorylase activity was demonstrated in bag fibres as well as in chain fibres. No differences could be detected in adenosine desaminase activity between these various intrafusal fibres. In the outer capsule of muscle spindles, high amount of type III collagen and elastin, but only small amount of type I collagen and fibronectin could be observed.

Adenosine Deaminase