Age-Related Alterations of Hyaluronan and Collagen in Extracellular Matrix of the Muscle Spindles

Background: Muscle spindles (MSs) play a crucial role in proprioception and locomotor coordination. Although the elasticity and viscosity of the extracellular matrix (ECM) within which MSs are embedded may play a key role in MS function, the impact of aging on ECM components is unclear. The aim of the current study was to investigate the age-related physiological changes of the ECM and to verify if these could be due to alterations of the environment directly surrounding MSs. Methods: Hematoxylin Eosin and picrosirius-red staining was carried out; collagen types I (COLI) and III (COLIII) were assessed, and biotinylated hyaluronan binding protein (HABP) immunohistochemical analysis was undertaken to evaluate alterations of the ECM in the intramuscular connective tissue (IMCT) of the hindlimbs of C57BL/6J male mice. Assessments were carried out on 6-week-old (Group A), 8-month-old (Group B), and 2-year-old (Group C) laboratory mice. Results: The capsule’s outer layer became progressively thicker with aging (it was 3.02 ± 0.26 μm in Group A, 3.64 ± 0.31 μm in Group B, and 5.81 ± 0.85 μm in Group C). The collagen in IMCT around and within the MSs was significantly higher in Group C, but there were no significant differences between Groups A and B. The MS capsules and continuous IMCT were primarily made up of COLI and COLIII. The average optical density (AOD) values of COLI in IMCT surrounding MS were significantly higher after aging (p < 0.05), but there were no significant differences in COLIII in the three groups (p > 0.05). HA was present in IMCT and filled the MSs capsule. The AOD of HABP of MS showed that there were lower HA levels in Group C with respect to Group A (p = 0.022); no significant differences were noted neither between Groups A and B nor between Groups B and C (p > 0.05). Conclusion: Age-related collagen accumulation and lower HA in the ECM in which the MSs were embedded may probably cause more stiffness in the ECM in vivo, which could help to partly explain the peripheral mechanisms underlying the age-related decline in functional changes related to MSs.

Enhanced Corticospinal Excitability in the Tibialis Anterior During Static Stretching of the Soleus in Young Healthy Individuals

Corticospinal excitability is known to be affected by afferent inflow arising from the proprioceptors during active or passive muscle movements. Also during static stretching (SS) afferent activity is enhanced, but its effect on corticospinal excitability received limited attention and has only been investigated as a single average value spread over the entire stretching period. Using transcranial magnetic stimulation (TMS) the present study was conducted to explore the time course of corticospinal excitability during 30 seconds SS.Motor evoked potentials (MEPs) after TMS were recorded from soleus (SOL) and tibialis anterior (TA) muscles in 14 participants during: a passive dynamic ankle dorsiflexion (DF), at six different time points during maximal individual SS (3, 6, 9, 18, 21 and 25 seconds into stretching), during a passive dynamic ankle plantar flexion (PF) and following SS.During passive DF, MEPs amplitude was greater than baseline in both TA and SOL (p=.001 and p=.005 respectively). During SS, MEPs amplitude was greater than baseline in TA (p=.006), but not in SOL. No differences between the investigated time points were found and no trend was detected throughout the stretching time. No effect in either muscle was observed during passive PF and after SS.These results could suggest that an increased activity of secondary afferents from SOL muscle spindles exert a corticomotor facilitation on TA. The muscle-unspecific response observed during passive DF could instead be attributed to an increased activation within the sensorimotor cortical areas as a result of the awareness of the foot passive displacements.

Ia EPSPs in rat spinal motoneurons are potentiated after a 5-week whole-body vibration

Whole-body vibration (WBV) is often applied as an alternative method for strength training or to prevent muscle force decrease. Previous studies indicated that WBV induced: 1) changes in the contractile parameters predominantly of fast motor units, 2) higher motoneuron excitability, and 3) higher motoneuron firing rates at lower stimulus intensities compared with the control. In this study, we evaluated the influence of WBV on Ia monosynaptic input from muscle spindles because the tonic vibration reflex is responsible for the enhancement of muscle activity observed after WBV. The aim was to answer the question of whether repeated activation of muscle spindles during WBV may result in altered synaptic excitation of motoneurons. WBV was performed on adult male Wistar rats, 5 days per week, for 5 weeks, and each daily session consisted of four 30-s runs of vibration at 50 Hz. Fast-type medial gastrocnemius motoneurons were investigated intracellularly in deeply anesthetized animals in the experimental (n=7, 34 motoneurons) and control (n=7, 32 motoneurons) groups. Monosynaptic Ia EPSPs were evoked by electrical stimulation of afferent fibers from the synergistic lateral gastrocnemius and soleus muscles. Data were analyzed using a mixed linear model. WBV induced an increase of the mean EPSP amplitude by 28% (P=0.025), correlated with the resting membrane potential and input resistance, and a shortening of the mean EPSP rise time by 11% (P=0.012). The potentiation of synaptic excitation of motoneurons indicates that WBV may support rehabilitation or training processes aimed at increasing muscle strength on the basis of increased motoneuronal drive.

Evaluating Sexual Dimorphism of the Muscle Spindles and Intrafusal Muscle Fibers in the Medial Gastrocnemius of Male and Female Rats

This study sought to investigate the sexual dimorphism of muscle spindles in rat medial gastrocnemius muscle. The muscles were cut transversely into 5–10 and 20 μm thick serial sections and the number, density, and morphometric properties of the muscle spindles were determined. There was no significant difference (p &gt; 0.05) in the number of muscle spindles of male (14.45 ± 2.77) and female (15.00 ± 3.13) rats. Muscle mass was 38.89% higher in males (1.08 vs. 0.66 g in females), making the density of these receptors significantly higher (p &lt; 0.01) in females (approximately one spindle per 51.14 mg muscle mass vs. one per 79.91 mg in males). There were no significant differences between the morphometric properties of intrafusal muscle fibers or muscle spindles in male and female rats (p &gt; 0.05): 5.16 ± 2.43 and 5.37 ± 2.27 μm for male and female intrafusal muscle fiber diameter, respectively; 5.57 ± 2.20 and 5.60 ± 2.16 μm for male and female intrafusal muscle fiber number, respectively; 25.85 ± 10.04 and 25.30 ± 9.96 μm for male and female shorter muscle spindle diameter, respectively; and 48.99 ± 20.73 and 43.97 ± 16.96 μm for male and female longer muscle spindle diameter, respectively. These findings suggest that sexual dimorphism in the muscle spindles of rat medial gastrocnemius is limited to density, which contrasts previous findings reporting differences in extrafusal fibers diameter.

Where is my hand in space? The internal model of gravity influences proprioception

Knowing where our limbs are in space is crucial for a successful interaction with the external world. Joint position sense (JPS) relies on both cues from muscle spindles and joint mechanoreceptors, as well as the effort required to move. However, JPS may also rely on the perceived external force on the limb, such as the gravitational field. It is well known that the internal model of gravity plays a large role in perception and behaviour. Thus, we have explored whether direct vestibular-gravitational cues could influence JPS. Participants passively estimated the position of their hand while they were upright and therefore aligned with terrestrial gravity, or pitch-tilted 45° backwards from gravity. Overall participants overestimated the position of their hand in both upright and tilted postures; however, the proprioceptive bias was significantly reduced when participants were tilted. Our findings therefore suggest that the internal model of gravity may influence and update JPS in order to allow the organism to interact with the environment.

Encoding of limb state by single neurons in the cuneate nucleus of awake monkeys

The cuneate nucleus (CN) is among the first sites along the neuraxis where proprioceptive signals can be integrated, transformed, and modulated. The objective of the study was to characterize the proprioceptive representations in CN. To this end, we recorded from single CN neurons in three monkeys during active reaching and passive limb perturbation. We found that many neurons exhibited responses that were tuned approximately sinusoidally to limb movement direction, as has been found for other sensorimotor neurons. The distribution of their preferred directions (PDs) was highly non-uniform and resembled that of muscle spindles within individual muscles, suggesting that CN neurons typically receive inputs from only a single muscle. We also found that the responses of proprioceptive CN neurons tended to be modestly amplified during active reaching movements compared to passive limb perturbations, in contrast to cutaneous CN neurons whose responses were not systematically different in the active and passive conditions. Somatosensory signals thus seem to be subject to a "spotlighting" of relevant sensory information rather than uniform suppression as has been suggested previously.

Ultrastructural characterization of peripheral denervation in a mouse model of Type III spinal muscular atrophy

Spinal muscular atrophy (SMA) is a heritable, autosomal recessive neuromuscular disorder characterized by a loss of the survival of motor neurons (SMN) protein, which leads to degeneration of lower motor neurons, and muscle atrophy. Despite SMA being nosographically classified as a motor neuron disease, recent advances indicate that peripheral alterations at the level of the neuromuscular junction (NMJ), involving the muscle, and axons of the sensory-motor system, occur early, and may even precede motor neuron loss. In the present study, we used a mouse model of slow progressive (type III) SMA, whereby the absence of the mouse SMN protein is compensated by the expression of two human genes (heterozygous SMN1A2G, and SMN2). This leads to late disease onset and prolonged survival, which allows for dissecting slow degenerative steps operating early in SMA pathogenesis. In this purely morphological study carried out at transmission electron microscopy, we extend the examination of motor neurons and proximal axons towards peripheral components, including distal axons, muscle fibers, and also muscle spindles. We document remarkable ultrastructural alterations being consistent with early peripheral denervation in SMA, which may shift the ultimate anatomical target in neuromuscular disease from the spinal cord towards the muscle. This concerns mostly mitochondrial alterations within distal axons and muscle, which are quantified here through ultrastructural morphometry. The present study is expected to provide a deeper knowledge of early pathogenic mechanisms in SMA.