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Capsule restricts the further study on muscle spindle function and the involved mechanism. The aim of this study was to establish the isolation method of intrafusal fibres from the isolated rat muscle spindle. Intrafusal fibres were harvested from muscle spindle of soleus muscle in rats using neutrase-collagenase digestion. A variety of incubation mediums have been tested to find out an appropriate medium of intrafusal fibers in vitro. Trypan blue staining was used to detect cell death, and patch clamp was used to record resting potential. The results showed that the intrafusal fibres incubated with amine acid-saline solution were almost all dead. DMEM could maintain good condition of the fibres, but excess CO2 ventilation would induce cellular swelling or even death. While Leiboviz's 15 (L-15) medium can guarantee 1-2 h of physiological condition of the intrafusal fibres. Coverslips treated with gelatin, polylysine and serum was the better interfaces for the intrafusal fibres to adhere easily, compared with regularly treated coverslip. The resting potential of intrafusal fibres was (-45.3 ± 5.1) mV, consistent with others obtained from in vivo muscle spindle from cats and frogs. These results suggest that the isolation method of the intrafusal fibres has been successfully established in the present study, providing a new approach in better understanding of muscle spindle activities and the involved mechanism.

One of the most important and urgent issues in the field of space medicine is to reveal the potential mechanism underlying the disused muscle atrophy during the weightlessness or microgravity environment. It will conduce to find out effective methods for the prevention and treatment of muscle atrophy during a long-term space flight. Increasing data show that muscle spindle discharges are significantly altered following the hindlimb unloading, suggesting a vital role in the progress of muscle atrophy. In the last decades, we have made a series of studies on changes in the morphological structure and function of muscle spindle following simulated weightlessness. This review will discuss our main results and related researches for understanding of muscle spindle activities during microgravity environment, which may provide a theoretic basis for effective prevention and treatment of muscle atrophy induced by weightlessness.

The aim of this study was to observe the electrophysiological characteristics of the isolated rat muscle spindle. The muscle spindle was isolated from rat soleus and the afferent discharge of the isolated muscle spindle was recorded by air-gap technique. In the basic physiological salt solution, the spontaneous impulses of muscle spindle were at a lower level with irregular intervals. The mean frequency of afferents was (51.78 ± 25.63) impulses/1 000 s (n = 13). The muscle spindle afferents were significantly increased and maintained over time by the addition of certain amino acids during the observation. The number of the action potential recorded per 1 000 s was 200-1 000 [mean: (687.62 ± 312.56) impulses/1 000 s, n = 17]. In addition to the typical propagated action potential, a large number of abortive spikes were observed. The results indicate that the activities of isolated muscle spindles in rats can be well maintained by the addition of certain amino acids. The results initially establish and provide the possibility for further research conducted in isolated rat muscle spindles.

The present study aimed to study the changes of neurotrophin-3 (NT-3) expression of intrafusal muscle fibers in rat soleus muscles under simulated weightlessness. The tail-suspension (SUS) rat model was used to simulate weightlessness. Forty mature female Sprague-Dawley rats were randomly assigned to ambulatory control (CON), 3-day SUS, 7-day SUS, 14-day SUS and 21-day SUS groups. Immunohistochemistry ABC staining method and enzyme linked immunosorbent assay (ELISA) were used to detect the NT-3 expression of intrafusal muscle fibers in rat soleus muscles. The results from the immunohistochemistry staining technique showed that the extrafusal muscle fibers did not exhibit the NT-3-like immunoreactivity, and NT-3-like immunoreactivity was mainly expressed in nuclear bag 1 and nuclear bag 2 fibers of the muscle spindles. The ELISA results showed that the expression quantity of NT-3 in rat soleus muscles in control, 3-day SUS, 7-day SUS, 14-day SUS and 21-day SUS groups were (14.23±1.65), (14.11±1.53), (13.09±1.47), (12.45±1.51) and (9.85±1.52) pg/mg of tissue respectively. Compared to the control group, the expression quantity of NT-3 was significantly decreased after 14 days of SUS (P<0.05). After 21 days of SUS, the NT-3 expression was further reduced (P<0.01). These results suggest that simulated weightlessness induces an obvious decrease in the NT-3 expression level of intrafusal fibers in rat soleus muscles. Accompanying the simulated weightlessness extension, NT-3 expression in rat soleus muscle spindles is progressively decreased. These changes may contribute to the proprioceptive adaptations to microgravity.

During hind limb unloading (HU), the soleus is often in a shortened position and the natural physiological stimulus of muscle spindles is altered, such that muscle spindle activity also changes. Using isolated spindle conditions, the present study investigates the electrophysiological activity and ultrastructure of muscle spindles following HU. Results show that muscle spindle discharges fall into either of two main patterns, single spikes or spike clusters in shortened positions, with a steady frequency of 18-38 spikes/s (mean 29.08 +/- 2.45) in an extended position. Following 14-day HU, afferent discharge activity was significantly altered in soleus muscle spindles. Duration of individual spikes was significantly prolonged, from 0.54 +/- 0.05 ms for control rats to 1.53 +/- 0.25 ms for rats in the HU group. In a shortened position, regular rhythm afferent discharges were obviously depressed, and the majority of muscle spindles became silent, while in an extended position, the discharges remained continuous but with decreased frequency. Results also show that the ultrastructure of muscle spindles experience degenerative changes during HU. Altered muscle spindle afference could possibly modify the activity of motor neurons and further affect the activity of extrafusal fibers.

To date, the medium and long-term space flight is urgent in need and has become a major task of our manned space flight program. There is no doubt that medium and long-term space flight has serious damaging impact upon human physiological systems. For instance, atrophy of the lower limb anti-gravity muscle can be induced during the space flight. Muscle atrophy significantly affects the flight of astronauts in space. Most importantly, it influences the precise manipulation of the astronauts and their response capacity to emergencies on returning to the atmosphere from space. Muscle atrophy caused by weightlessness may also seriously disrupt the normal life and work of the astronauts during the re-adaptation period. Here we summarize the corresponding research concentrating on weightlessness-induced changes of muscular structure and function. By combining research on muscle pain, which is a common clinical pain disease, we further provide a hypothesis concerning a dynamic feedback model of "weightlessness condition right triple arrow muscular atrophy <--> muscle pain". This may be useful to explore the neural mechanisms underlying the occurrence and development of muscular atrophy and muscle pain, through the key study of muscle spindle, and furthermore provide more effective therapy for clinical treatment.

Objective: To shed some light on treatment of muscle atrophy through testing the effects of several Chinese traditional drugs that can invigorate blood circulation on the afferent discharge of muscle spindle., Method: Single muscle spindle was isolated from sartorius muscle of toad. Afferent discharge from the muscle spindle was recorded using air-gap technique and effects of three Chinese traditional drugs on afferent activity of the muscle spindle were observed., Result: Ligustrazine could markedly tingle the spontaneous electrical activity of the muscle spindle, and it also showed a good dose-response relationship, while Salvia Miltiorrhiza and Safflower had no such excitative effect., Conclusion: It is known that ligustrazine can expand blood vessels and agitate microcirculation, and this effect on the muscle spindle makes it helpful for persons with muscle atrophy.

Objective: To study influence of 100 Hz sinusoidal vibration on muscle spindle afferents discharges of rat soleus muscles in simulated-weightlessness situation., Method: The tail-suspended rat model was used to simulate weightlessness, and 100 Hz sinusoidal vibration was performed by a vibrator. Unit activity was recorded electrophysiologically from the centrally cut filaments of the spinal dorsal roots innervating muscle spindles of the rat soleus muscle; then observation on the changes in afferent discharges from muscle spindle in a rat soleus muscles were made after 7 d., Result: 1) Compared with control muscle spindle afferents discharges of rat soleus muscles decreased after 7 d tail-suspension (P<0.05). 2) Compared with suspension the afferent discharges from soleus muscle spindles increased after 7 d tail-suspension with 100 Hz sinusoidal vibration., Conclusion: Sinusoidal vibration (100 Hz; 300 micrometers) can selectively activate muscle spindles. Muscle spindle afferent input of the suspended rats increased after the application of 7 d 100 Hz sinusoidal vibration.

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.

Objective: To examine the myosin heavy chain expression in intrafusal fibers in rat soleus muscle before and after simulated weightlessness and to analyse the mechanism involved in muscle spindle deterioration during simulated weightlessness., Method: The rats were divided into 3 groups: 7 d, 14 d tail-suspension group and control group. Control and experimental spindles were examined using monoclonal antibodies specific for myosin heavy chains of slow-tonic (ALD58) and fast-twitch (MF30) chicken muscles by the immunoperoxidase reaction utilizing the ABC (avidin-biotin-complex) method., Result: The extrafusal muscle fibers did not exhibit immunoreactivity to ALD58 and MF30. Only intrafusal fibers bound these antibodies. The bag 1 fiber and bag 2 fiber reacted to ALD58. The bag 2 fiber and chain fibers stained with MF30. After 7 d suspension the immunoreactivity of the bag 1 fiber and bag 2 fiber to ALD58 decreased, while the immunoreactivity of the bag 2 fiber and chain fibers to MF30 increased. After 14 d suspension the immunoreactivity of the bag 1 fiber and bag 2 fiber to ALD58 decreased remarkably, even disappeared. While all of the intrafusal fibers reacted to MF30 and the immunoreactivity increased obviously., Conclusion: The expression level and patterns of MHC in intrafusal fibers of rat soleus muscle changed after simulated weightlessness.

Objective: To study changes in structure and innervation of soleus muscle spindle in rats induced by simulated weightlessness., Method: Weightlessness was simulated by tail suspension in female rats. The technique of Fawosky's Staining was used to detect the morphology of nerve endings of soleus muscle spindle in 7 d, 14 d, 21 d tail-suspended and control rats., Result: The intrafusal muscle fibers were regular and nerve endings were intact and clear in the soleus spindles of control rats. No obvious change was observed in muscle spindles of 7 d tail-suspended rats. Intrafusal muscle fibers became irregular, with rough and vague nerve endings in 14 d tail-suspended rats. The distinct retrograde change of muscle spindles and nerve endings were observed in 21 d tail-suspended rats. Degeneration and rupture were found in nerve endings, which made it unclear and pale in staining., Conclusion: Simulated weightlessness could induce time-related changes in structure and innervation of soleus muscle spindle in rats.

Objective. To analyze the mechanism involved in muscle spindle deterioration during simulated weightlessness. Method. Using the immunoperoxidase reaction utilizing the ABC (avidin-biotin-complex) method, Calbindin D28K-like immunoreactivity (CaBP-LI) of intrafusal fibres in soleus muscle were detected in 7 d, 14 d tail-suspended rats and compared with control rats. Result. The extrafusal muscle fibres and nerve fibres did not exhibit immunoreactivity to CaBP. CaBP-LI was found in some of the intrafusal muscle fibres in all the muscle spindles. After 14 d suspension, the immunoreactivity to CaBP of the intrafusal fibres decreased markedly. Conclusion. Simulated weightlessness could induce changes in the immunoreactivity of rat soleus muscle spindle to CaBP, which may contribute to the deterioration of muscle spindle.

Objective: To study changes in ultrastructure of soleus muscle spindle induced by simulated weightlessness in rats., Method: Weightlessness was simulated by tail suspension in female rats. The ultrastructure of isolated soleus muscle spindle were observed in 4 d, 7 d, 14 d tail-suspended and 14 d recovered rats., Result: Chaotic myofibril, hyperplasia of mitochondria, and enlargement of terminal cisterna were found in the ultrastructure of isolated soleus muscle spindle in 4 d tail-suspended rats. The ultrastructure of isolated soleus muscle spindle in 7 d tail-suspended rats showed more distinct changes, while that in 14 d tail-suspended rats showed obvious retrograde changes. The ultrastructure of isolated muscle spindle in 14 d recovered rats after 14 d tail-suspension were similar to that of control group., Conclusion: Simulated weightlessness could induce time-related and reversible changes in ultrastructure of soleus muscle spindle in rats.

It is well known that the muscle spindle is a receptor of muscle's tension and length, it plays an important role in maintaining the muscle's tension. The aim of the present study is to compare the cross-section area (CSA) and the immunoreactivity of conjugated-ubiquitin in soleus extrafusal and intrafusal fibers after simulated-microgravity in order to demonstrate the role of muscle spindle in muscle atrophy induced by simulated microgravity.

The static component of the spindle potential provoked during stretch of isolated muscle spindles of the frog was reversed during the application of depolarizing currents ranging from 0.2 to 5 nA in normal Ringer solution and also in Na+-free Ringer solution. In the same range of current intensities, spontaneous rhythmic hyperpolarizations due to [Ca2+]i-activated GK, an attenuation of membrane impedance, and an anomalous decrease in amplitude of the afferent spikes were observed. All 4 phenomena were abolished by K+ channel blockers (10 mM CsCl, 1-2 mM 4-aminopyridine (4-AP), or 20 mM tetraethylammonium chloride (TEA], Ca2+ channel blockers (5-10 mM CoCl2, MnCl2, 1-2 mM CdCl2 or 0.5 mM verapamil) or 0.1 mM quinine. The amplitude of the static component of the spindle potential was markedly increased at threshold concentration of the K+ channel blockers (5 mM CsCl, 0.1-0.5 mM 4-AP or 5-10 mM TEA), but the component disappeared at that of the Ca2+ channel blockers. The rhythmic hyperpolarizations are associated with the spindle potential, except for its dynamic component, which often triggers a hyperpolarizing deflection. We suggest that both the static component of the spindle potential and rhythmic hyperpolarizations are due to GK(Ca) in the intracapsular axon, either along the terminal or at the branching nodes, or both; and that the receptor potential contributes to, but is not the same as, the spindle potential.