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Neuregulin induces the expression of transcription factors and myosin heavy chains typical of muscle spindles in cultured human muscle.

Neuregulin (NRG) (also known as ARIA, GGF, and other names) is a heparin sulfate proteoglycan secreted into the neuromuscular junction by innervating motor and sensory neurons. An integral part of synapse formation, we have analyzed NRG-induced changes in gene expression over 48 h in primary human myotubes. We show that in addition to increasing the expression of acetylcholine receptors on the myotube surface, NRG treatment results in a transient increase of several members of the early growth response (Egr) family of transcription factors. Three Egrs, Egr1, -2, and -3, are induced within the first hour of NRG treatment, with Egr1 and -3 RNA levels showing the most significant increases of approximately 9- and 16-fold, respectively. Also noted was a corresponding increase in protein levels for both of these transcription factors. Previous literature indicates that Egr3 expression is required for the formation of muscle spindle fibers, sensory organs that are distinct from skeletal muscle contractile fibers. At the molecular level, muscle spindle fibers express a unique subset of myosin heavy chains. Two isoforms of the myosin heavy chain, the slow development and neonatal, were found to be increased in our myotube cultures after 48 h of treatment with NRG. Taken together, these results indicate that not only can NRG induce the expression of a transcription factor key to spindle fiber development (Egr3), but that a portion of this developmental process can be replicated in vitro.

Agrin↗

Muscle spindles in the digastric muscle of the rabbit.

The digastric muscle of the rabbit was studied for the presence of muscle spindles. One of the 19 adult digastric muscles we examined contained in excess of 12 spindles scattered throughout the belly. No spindles were found in the contralateral muscle, or in any of the digastric muscles from other animals. Although jaw depressor muscles of most mammals contain few, if any, muscle spindles, their occasional presence suggests that these muscles have a potential for spindle formation.

Animals↗

Immunohistochemical localization of alpha(1a)-adrenoreceptors in muscle spindles of rabbit masseter muscle.

The expression of alpha(1a)-adrenoreceptors (alpha(1a)-ARs) within the muscle spindles of rabbit masseter muscle was investigated. The alpha(1a)-ARs were detected by immunohistochemical fluorescent method and examined along the entire length of 109 cross serially sectioned spindles. The sympathetic fibers were visualized by the immunofluorescent labeling of the noradrenaline synthesizing enzymes tyrosine hydroxylase (TH) and dopamine beta-hydroxylase (DBH). In order to recognize the intrafusal muscle fiber types, antibodies for different myosin heavy chain isoforms (MyHCI) were used. TH and DBH immunolabeled nerve fibers have been observed within the capsule lamellar layers, in the periaxial fluid space and close to intrafusal muscle fibers. The alpha(1a)-ARs were detected on the smooth muscle cells of the blood vessels coursing in the muscle and in the capsule lamellar layers or within the periaxial fluid space of the spindles. Moreover, at the polar regions of a high percentage (88.1%) of muscle spindles a strong alpha(1a)-ARs immunoreactivity was present on the intrafusal muscle fibers. In double immunostained sections for alpha(1a)-ARs and MyHCI it was evidenced that both bag, and nuclear chain fibers express alpha(1a)-ARs. The receptors that we have detected by immunofluorescence may support a direct control by adrenergic fibers on muscle spindle.

Animals↗

Functional morphological interpretation of the distribution of muscle spindles in the jaw muscles of the mallard (Anas platyrhynchos).

The morphology and distribution of muscle spindles of jaw and tongue muscles in the mallard were examined in serial transverse sections of single muscles and in horizontal sections of a whole head. Our observations on spindle morphology are in agreement with previous descriptions of spindles in birds. Some spindles differ in their innervation and the pattern of intrafusal muscle fibers. The spindles of individual adductor and pterygoid muscles are distributed unevenly. Some adductor muscles lack spindles, whereas those of other muscles are confined to limited areas. Jaw opening muscles and extrinsic tongue muscles lack spindles. The stretch of extrafusal muscle fibers could be estimated from the difference in sarcomere length for birds with the beak open and closed. Not all muscle fiber groups are stretched evenly over the whole range of jaw opening. Only those fiber groups that are continuously stretched during jaw opening contain spindles.

Animals↗

An ultrastructural study of nerve terminal degeneration in muscle spindles of the tenuissimus muscle of the cat.

Muscle spindles in the tenuissimus muscle of the cat were studied between 12 and 168 h after cutting or freezing the nerve to this muscle. Degenerative changes in sensory and motor nerve terminals on intrafusal muscle fibres were observed using the electron microscope. Comparisons were made with spindles from unoperated or sham-operated cats. The earliest degenerative changes were seen in sensory and motor terminals at 20-24 h after the lesion. No nerve endings were seen by 114 h after denervation. The most consistent initial signs of degeneration were: (1) the presence of abnormal mitochondria and dense bodies in sensory terminals, and (2) a decrease in the number and clumping of synaptic vesicles combined with an increase in glycogen and neurofilaments in motor endings. Intrafusal fibres participate in the removal of degenerating sensory endings. Schwann cells phagocytose degenerating motor terminals. The disappearance of nerve terminals precedes the complete degeneration of preterminal myelinated fibres within the muscle spindle.

Animals↗

Roles of muscle activity and load on the relationship between muscle spindle length and whole muscle length in the freely walking cat.

The objective of this research was to compare the length of muscle spindles to the length of the whole muscle, during normal movements. Pairs of piezoelectric crystals were implanted near the origin and insertion of muscle fibres in the medial gastrocnemius (MG) muscle of cats. The distance between crystals was measured with pulsed ultrasound, the origin-to-insertion length of the MG muscle was measured with a transducer made of saline-filled silicone tubing, MG force was measured with a tendon force transducer and EMG activity was selectively recorded in the vicinity of implanted crystals. These signals were simultaneously recorded during posture or locomotion on a motorized treadmill. Three periods were identified in the step cycle, during which the relation between muscle length and spindle length changed dramatically. In period I (roughly corresponding to the late F and E1 phases of swing), the MG muscle and spindles followed similar length changes: both were stretched and then shortened by about 6 mm. In period II (corresponding to the stance phase, E2-E3) the MG muscle yielded under the weight of the body and was stretched by 1-3 mm, whereas the MG spindles typically continued shortening. In period III, the MG muscle shortened rapidly by 6-8 mm after the foot left the ground and then stretched again by about the same amount, whereas the spindles could remain nearly isometric. We attribute these large discrepancies in muscle and spindle length to the architecture of the MG muscle and the compliance of long tendinous elements in series with the spindles. We conclude that the length changes imposed on muscle spindles during voluntary movements are not simply related to the parent muscle length changes and cannot be estimated without taking into account the muscle architecture, the location of the spindle within the muscle, the level of muscle activation and the external load.

Animals↗

An allometric analysis of the number of muscle spindles in mammalian skeletal muscles.

An allometric analysis of the number of muscle spindles in relation to muscle mass in mammalian (mouse, rat, guinea-pig, cat, human) skeletal muscles is presented. It is shown that the trend to increasing number as muscle mass increases follows an isometric (length) relationship between species, whereas within a species, at least for the only essentially complete sample (human), the number of spindles scales, on average, with the square root rather than the cube root of muscle mass. An attempt is made to reconcile these apparently discrepant relationships. Use of the widely accepted spindle density (number of spindles g(-1) of muscle) as a measure of relative abundance of spindles in different muscles is shown to be grossly misleading. It is replaced with the residuals of the linear regression of ln spindle number against ln muscle mass. Significant differences in relative spindle abundance as measured by residuals were found between regional groups of muscles: the greatest abundance is in axial muscles, including those concerned with head position, whereas the least is in muscles of the shoulder girdle. No differences were found between large and small muscles operating in parallel, or between antigravity and non-antigravity muscles. For proximal vs. distal muscles, spindles were significantly less abundant in the hand than the arm, but there was no difference between the foot and the leg.

Adult↗

Differences in muscle spindle structure between pigeon muscles used in aerial and terrestrial locomotion.

Muscle spindle density (number of spindles per gram of muscle) of all 29 muscles of the forearm and leg of the domestic pigeon was evaluated by counting receptors in van Gieson-stained serial cross sections. Extra- and intra-fusal fiber-type profiles were determined from histochemical preparations. Muscles of the leg had on the average significantly more avian slow-twitch oxidative extrafusal fibers (22.5 vs. 0.8%) and slower contraction times than muscles of the forearm, but fiber-type profiles and gross actions of muscles showed no consistent relation to the relative abundance of receptors. Differences in intrafusal fiber-type composition among spindles were sought because of their potential effect on the quality of the afferent discharge. The number of intrafusal fibers per spindle was on the average significantly less (4.57 vs 5.99) in the muscles of the leg than in those of the forearm; and of spindles with the same number of intrafusal fibers, those in the leg had smaller periaxial spaces. Distribution of intrafusal fiber types identified with the myofibrillar adenosine triphosphatase reaction differed among spindles of varying sizes. An acid- and alkali-labile type occurred most frequently (P = 0.05) in spindles with one to three intrafusal fibers, and an acid-labile and alkali-stable type was most often seen (P = 0.05) in spindles with 4 to 7 intrafusal fibers. The smaller receptors were more abundant in the leg, while the larger ones were about equally distributed between the two extremities. Muscle fibers with dimensions that sometimes approached small extrafusal fibers were present in about 3% of the axial bundles examined, most of them in the forearm. The selective morphological variation of avian muscle spindles may represent the structural basis for qualitatively different afferent discharges that relate to the characteristic types of locomotion served by the two extremities.

Animals↗

Capsules of duck muscle spindles.

Duck muscle spindles show a large outer capsule enclosing a number of intrafusal muscle fibres which are individually encircled by an inner capsule. The outer capsule consists of a multilayer of 4-6 flattened cells with elongated nuclei, and are usually lined by a basement membrane. They resemble and are linked with perineural cells of the nerve bundle that innervates the spindle. There is overlapping and interdigitation between branching processes of these cells. Some apposing surfaces develop junctional complexes in the form of hemi-desmosomes and desmosomes. The cytoplasmic content shows numerous pinocytotic vesicles, some polyribosomes, lamellated cisternae of rough endoplasmic reticulum and microfilaments. The inner capsule consists of cells showing, at the mid-equatorial region, a large ovoid nucleus and extensive branching of thin and long processes that overlap, forming numerous layers around each intrafusal muscle fibre. Bundles of collagen fibrils in a crescent-shaped configuration occur circumferentially between the multilayer of processes and the muscle fibre. Some linkages between the inner and the outer capsule cell processes provide a network that subdivides the large periaxial space into compartments. There is no basement material lining the inner capsule cells and the processes. Some cytoplasmic area occurs around the ovoid nucleus and the cytoplasm varies in density, showing some dilated short profiles of rough endoplasmic reticulum, a few pinocytotic vesicles and microfilaments.

Animals↗

An electron microscopic study of the muscle spindle in the arytenoid muscle of the human larynx.

We used transmission electron microscopy to examine a muscle spindle found in serial sections of the arytenoid muscle of the human larynx. Eight intrafusal fibers were observed at the equatorial region and contained two types of muscle fibers, nuclear bag fibers and nuclear chain fibers. The sensory nerve endings found on the nuclear bag fibers were annular or varicose in shape and contained numerous mitochondria. Another small sensory ending was composed of many vesicles and some dense granules. The fusimotor nerve endings found on the nuclear chain fibers had three types of endings with different sizes and complexities.

Humans↗

Influences on the gamma-muscle-spindle system from contralateral muscle afferents stimulated by KCl and lactic acid.

The aim of the present study was to investigate whether increased concentrations of lactic acid and potassium chloride (KCl) in contralateral muscles can influence the sensitivity of primary and secondary muscle spindle afferents (MSAs) from ipsilateral extensor and flexor muscles. The experiments were performed on 7 cats anaesthetised with alpha-chloralose. Recordings were made simultaneously from 2-12 single MSAs from the triceps surae (GS) and/or the posterior biceps and semitendinosus muscles (PBSt). The mean rate of firing and the amplitude of a fitted sine curve of MSA responses to sinusoidal stretching of the receptor bearing muscles were determined. Responses of 42 primary MSAs (17 from PBSt and 25 from GS) were recorded. On 33 of these, clear-cut alterations in sinusoidal response were evoked by injection of 1 ml KCl (200-400 mM) or 1 ml lactic acid (20-50 mM) into the arterial supply of the contralateral GS or PBSt muscles. Six out of 8 secondary MSAs showed sizeable effects to increased intramuscular concentrations of KCl and/or lactic acid (3 from PBSt and 3 from GS). On both primary and secondary MSAs, from GS as well as from PBSt muscles, the large majority of effects were excitatory. All effects on secondary MSAs were compatible with reflex actions on static fusimotor neurones, whereas on primary MSAs different types of reflex responses were observed (i.e. pure static, pure dynamic and mixtures of static and dynamic fusimotor actions). Stimuli related alterations in MSA responses were completely abolished when the contralateral GS or PBSt nerves were anaesthetised.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Characteristics of the muscle spindle endings of the masticatory muscles in the rabbit under halothane anesthesia.

To explore the response characteristics of muscle spindle units in the masticatory muscles in the rabbit, the responses of muscle spindle units were recorded from the mesencephalic trigeminal nucleus (MesV) under halothane anesthesia during ramp-and-hold stretches. Three firing patterns, initial burst (IB) at the onset of the dynamic phase, negative adaptation (NA) at the end of the dynamic phase and firing during the release (FDR) phase, were observed during muscle stretch. IB was present at higher stretch velocities, FDR at lower stretch velocities. The velocity at which an IB or FDR was present was different from unit to unit. Because, within the range of the velocities of stretch tested, units with NA always showed NA and units without NA never did, all recorded units were divided into two groups on the basis of the existence of NA (NA(+) or NA(-) units). Response characteristics of the two groups were then compared. NA(+) units showed an IB more frequently and FDR less frequently than NA(-) units. NA(+) units had significantly higher dynamic responsiveness and discharge variability than NA(-) units. The conduction velocity of the afferents of NA(+) units was higher than that of NA(-) units. However, distributions of these measurements were not bimodal. These results suggest that NA is the useful criteria to classify the muscle spindle endings in the masticatory muscles in the rabbit under halothane anesthesia.

Anesthesia↗

Effects of tail suspension on electrophysiological characteristics of muscle spindles in isolated rat soleus muscle.

OBJECTIVE: To study changes in electrophysiological characteristics of muscle spindles induced by simulated weightlessness in isolated rats soleus. METHOD: Weightlessness was simulated by tail-suspension in female rats. Using electro-physiological technique, the spontaneous discharge of muscle spindle and its response to ramp-and-hold stretch were observed in isolated soleus of 7 d, 14 d tail-suspended and control rats. RESULT: Soleus muscle spindles of rat manifest a sharp decrease in spontaneous discharge frequency and the response to ramp-and-hold stretch after 7 d tail-suspension. More significant changes were observed in rats after 14 d tail-suspension. CONCLUSION: Simulated weightlessness could induce time-related changes in electrophysiological characteristics of soleus muscle spindles in rat.

Animals↗

Inhibition of muscle spindle afferent activity during masseter muscle fatigue in the rat.

The influence of muscle fatigue on the jaw-closing muscle spindle activity has been investigated by analyzing: (1) the field potentials evoked in the trigeminal motor nucleus (Vmot) by trigeminal mesencephalic nucleus (Vmes) stimulation, (2) the orthodromic and antidromic responses evoked in the Vmes by stimulation of the peripheral and central axons of the muscle proprioceptive afferents, and (3) the extracellular unitary discharge of masseter muscle spindles recorded in the Vmes. The masseter muscle was fatigued by prolonged tetanic masseter nerve electrical stimulation. Pre- and postsynaptic components of the potentials evoked in the Vmot showed a significant reduction in amplitude following muscle fatigue. Orthodromic and antidromic potentials recorded in the Vmes also showed a similar amplitude decrease. Furthermore, muscle fatigue caused a decrease of the discharge frequency of masseter muscle spindle afferents in most of the examined units. The inhibition of the potential amplitude and discharge frequency was strictly correlated with the extent of muscle fatigue and was mediated by the group III and IV afferent muscle fibers activated by fatigue. In fact, the inhibitory effect was abolished by capsaicin injection in the masseter muscle that provokes selective degeneration of small afferent muscle fibers containing neurokinins. We concluded that fatigue signals originating from the muscle and traveling through capsaicin-sensitive fibers are able to diminish the proprioceptive input by a central presynaptic influence. In the second part of the study, we examined the central projection of the masseter small afferents sensitive to capsaicin at the electron-microscopic level. Fiber degeneration was induced by injecting capsaicin into the masseter muscle. Degenerating terminals were found on the soma and stem process in Vmes and on the dendritic tree of neurons in Vmot. This suggests that small muscle afferents may influence the muscle spindle activity through direct synapses on somata in Vmes and on dendrites of neurons in Vmot.

Action Potentials↗

Intrafusal fiber type composition of muscle spindles in the first human lumbrical muscle.

We studied muscle spindles in the first lumbrical muscle of adult humans using myofibrillar ATPase (mATPase) activity. We found that muscle spindles exhibited a marked variability with respect to the number, position, length and detailed histochemical features of nuclear bag1, nuclear bag2 and nuclear chain fibers. Regarding mATPase activity, the nuclear bag2 fibers displayed lower alkali-stable mATPase activity along their length and many nuclear bag1 fibers tended to have lower acid-stable activity in the outer B region, whereas nuclear chain fibers exhibited medium acid-stable mATPase activity at pH 4.6. Almost 10% of spindle fibers displayed atypical features, as they were either located only at one spindle pole or exhibited mixed characteristics at either pole. The number of intrafusal fibers per spindle varied between 8 and 24. Strikingly, only 2 pairs from 22 muscle spindles had identical allotments of their intrafusal fibers. Muscle spindles in the first human lumbrical muscle contained more intrafusal fibers (12.3 +/- 4 per spindle on average) and especially relatively more nuclear bag fibers compared to other human skeletal muscles. Since each spindle apparently represents a unique morphological and physiological entity, the observed variability in the number and characteristics of intrafusal fibers in the first human lumbrical muscle likely reflects a wide range of finely tuned muscle spindle responses.

Adenosine Triphosphatases↗

Responses of muscle spindles in feline dorsal neck muscles to electrical stimulation of the cervical sympathetic nerve.

Previous studies performed in jaw muscles of rabbits and rats have demonstrated that sympathetic outflow may affect the activity of muscle spindle afferents (MSAs). The resulting impairment of MSA information has been suggested to be involved in the genesis and spread of chronic muscle pain. The present study was designed to investigate sympathetic influences on muscle spindles in feline trapezius and splenius muscles (TrSp), as these muscles are commonly affected by chronic pain in humans. Experiments were carried out in cats anesthetized with alpha-chloralose. The effect of electrical stimulation (10 Hz for 90 s or 3 Hz for 5 min) of the peripheral stump of the cervical sympathetic nerve (CSN) was investigated on the discharge of TrSp MSAs (units classified as Ia-like and II-like) and on their responses to sinusoidal stretching of these muscles. In some of the experiments, the local microcirculation of the muscles was monitored by laser Doppler flowmetry. In total, 46 MSAs were recorded. Stimulation of the CSN at 10 Hz powerfully depressed the mean discharge rate of the majority of the tested MSAs (73%) and also affected the sensitivity of MSAs to sinusoidal changes of muscle length, which were evaluated in terms of amplitude and phase of the sinusoidal fitting of unitary activity. The amplitude was significantly reduced in Ia-like units and variably affected in II-like units, while in general the phase was affected little and not changed significantly in either group. The discharge of a smaller percentage of tested units was also modulated by 3-Hz CSN stimulation. Blockade of the neuromuscular junctions by pancuronium did not induce any changes in MSA responses to CSN stimulation, showing that these responses were not secondary to changes in extrafusal or fusimotor activity. Further data showed that the sympathetically induced modulation of MSA discharge was not secondary to the concomitant reduction of muscle blood flow induced by the stimulation. Hence, changes in sympathetic outflow can modulate the afferent signals from muscle spindles through an action exerted directly on the spindles, independent of changes in blood flow. It is suggested that such an action may be one of the mechanisms mediating the onset of chronic muscle pain in these muscles in humans.

Adrenergic alpha-Antagonists↗