Effects of low-intensity exercise on contractile property of skeletal muscle and the number of motor neurons in diabetic rats.

The mode of diabetes-induced muscle and motor neuron damage depends on the type of muscle and motor neuron. One of the purposes of exercise therapy for diabetes is to improve blood glucose levels; however, information on the effects of low-intensity exercise on muscle and motor neuron disorders remain unknown. Therefore, this study aimed to examine the effects of low-intensity exercise on diabetes-induced muscle and motor neuron damage in a rat model of type 1 diabetes mellitus. We subjected adult male Wistar rats treated with streptozotocin to develop type 1 diabetes and age-matched rats to low-intensity treadmill exercise for 12 weeks. We recorded electrically evoked maximum twitch tension in leg muscles, and examined the number of motor neurons and cell body sizes. Low-intensity exercise ameliorated the prolonged half-relaxation time and the decreased numbers of the retrograde-labeled motor neurons observed in the soleus muscle of type 1 diabetic rats. However, no effect was observed in the diabetic group, as atrophy was not improved and the twitch force in the medial gastrocnemius muscle was decreased in the diabetic group. In addition, there was no improvement in the blood glucose levels after exercise. These data indicate that low-intensity exercise may relieve the onset of muscle and motor neuron damage in the soleus muscle of type 1 diabetic rats.

Motor skills training-induced activation of descending pathways mediating cortical command to hindlimb motoneurons in experimental diabetic rats.

Diabetes disrupts the corticospinal tract (CST) system components that control hindlimb and trunk movement, resulting in weakness of the lower extremities. However, there is no information about a method to improve these disorders. This study aimed to investigate the rehabilitative effects of 2 weeks of aerobic training (AT) and complex motor skills training (ST) on motor disorders in streptozotocin-induced type 1 diabetic rats. In this study, electrophysiological mapping of the motor cortex showed that the diabetes mellitus (DM)-ST group had a larger motor cortical area compared to the DM-AT group and sedentary diabetic animals. Moreover, hand grip strength and rotarod latency increased in the DM-ST group; however, these two parameters did not change in the DM-AT group, as well as in control and sedentary diabetic rats. Furthermore, in the DM-ST group, cortical stimulation-induced and motor-evoked potentials were preserved after the interception of the CST; however, this potential disappeared after additional lesions were made on lateral funiculus, suggesting that their function extends to activating motor descending pathways other than the CST locating lateral funiculus. According to immunohistochemical analysis, the larger fibers present on the dorsal part of the lateral funiculus, which corresponds to the rubrospinal tract of the DM-ST group, expressed the phosphorylated growth-associated protein, 43 kD, which is a specific marker of axons with plastic changes. Additionally, electrical stimulation of the red nucleus revealed expansion of the hindlimb-responsible area and increased motor-evoked potentials of the hindlimb in the DM-ST group, suggesting a strengthening of synaptic connections between the red nucleus and spinal interneurons driving motoneurons. These results reveal that ST induces plastic changes in the rubrospinal tract in a diabetic model, which can compensate for diabetes by disrupting the CST system components that control the hindlimb. This finding suggests that ST can be a novel rehabilitation strategy to improve motor dysfunctions in diabetic patients.

Functional and Structural Changes in the Corticospinal Tract of Streptozotocin-Induced Diabetic Rats.

This study aimed to reveal functional and morphological changes in the corticospinal tract, a pathway shown to be susceptible to diabetes. Type 1 diabetes was induced in 13-week-old male Wistar rats administered streptozotocin. Twenty-three weeks after streptozotocin injection, diabetic animals and age-matched control animals were used to demonstrate the conduction velocity of the corticospinal tract. Other animals were used for morphometric analyses of the base of the dorsal funiculus of the corticospinal tract in the spinal cord using both optical and electron microscopy. The conduction velocity of the corticospinal tract decreased in the lumbar spinal cord in the diabetic animal, although it did not decrease in the cervical spinal cord. Furthermore, atrophy of the fibers of the base of the dorsal funiculus was observed along their entire length, with an increase in the g-ratio in the lumbar spinal cord in the diabetic animal. This study indicates that the corticospinal tract fibers projecting to the lumbar spinal cord experience a decrease in conduction velocity at the lumbar spinal cord of these axons in diabetic animals, likely caused by a combination of axonal atrophy and an increased g-ratio due to thinning of the myelin sheath.

Sodium-glucose co-transporter (SGLT) inhibitor restores lost axonal varicosities of the myenteric plexus in a mouse model of high-fat diet-induced obesity.

Diabetes impairs enteric nervous system functions; however, ultrastructural changes underlying the pathophysiology of the myenteric plexus and the effects of sodium-glucose co-transporter (SGLT) inhibitors are poorly understood. This study aimed to investigate three-dimensional ultrastructural changes in axonal varicosities in the myenteric plexus and the effect thereon of the SGLT inhibitor phlorizin in mice fed a high-fat diet (HFD). Three-dimensional ultrastructural analysis using serial block-face imaging revealed that non-treated HFD-fed mice had fewer axonal varicosities and synaptic vesicles in the myenteric plexus than did normal diet-fed control mice. Furthermore, mitochondrial volume was increased and lysosome number decreased in the axons of non-treated HFD-fed mice when compared to those of control mice. Phlorizin treatment restored the axonal varicosities and organelles in HFD-fed mice. Although HFD did not affect the immunolocalisation of PGP9.5, it reduced synaptophysin immunostaining in the myenteric plexus, which was restored by phlorizin treatment. These results suggest that impairment of the axonal varicosities and their synaptic vesicles underlies the damage to the enteric neurons caused by HFD feeding. SGLT inhibitor treatment could restore axonal varicosities and organelles, which may lead to improved gastrointestinal functions in HFD-induced obesity as well as diabetes.

Motor cortex somatotopic presentation after restriction of neck movement in rats.

[Purpose] In this study, we aimed to investigate the effects of neck movement restriction on somatotopic mapping of the motor cortex. We restricted cervical extension for two weeks and investigated the effects on motor cortex somatic representation in rats. [Subjects and Methods] We placed six Wistar rats into each of three groups: (i) the experimental group, in which cervical extension was restricted; (ii) the sham group, in which cervical movement was not restricted, but a splint was placed in the shoulder girdle; and (iii) the control group. After cervical immobilization for two weeks, we evaluated the motor cortex somatic representation using intra-cortical micro-stimulation. [Results] In the experimental group, the areas of the cervical and vibrissal domains of the motor cortex decreased by approximately 50%, and the forelimb domain showed slight reduction. In addition, a trunk domain formed at the locus of the vibrissal area. There were no differences between the sham and control groups. [Conclusion] Restriction of cervical extension for two weeks resulted in changes in motor cortex somatic representation. Reversible changes occurred in cortical areas that controlled the neck and parts of the body involved in cervical movement.

Neck and trunk representations in the primary motor cortex in rats.

[Purpose] The neck and trunk play crucial roles in body movement and are extremely important areas of treatment for physical therapists. However, many aspects of the neural basis of this motor control remain unknown. Therefore, we investigated the distribution and electrophysiological properties of the neck and trunk in the primary motor cortex in rats. [Subjects and Methods] Using intracortical microstimulation, we investigated the somatotopic representation and movements induced by electrical stimulation of the neck and truck areas of the motor cortex in 8 Wistar rats. [Results] We determined that the neck and trunk areas are located separately on the rostral and caudal sides of the motor cortex, respectively. The neck area was significantly larger in size, while the threshold was significantly larger for the trunk area. Stimulation of the neck area with a current higher than the threshold induced movement of the forelimbs, jaw, trunk, and whiskers. However, stimulation of the trunk area did not result in movement in sites other than the trunk. [Conclusion] During movement, the respective activities of the neck and trunk are interdependent. However, due to the separate locations of these areas in the motor cortex, their properties differ greatly.

Effects of streptozotocin-induced diabetes on leg muscle contractile properties and motor neuron morphology in rats.

Skeletal muscle fiber subtypes are differentially sensitive to diabetes-related pathology; For example, fast-twitch muscles exhibit severe decreases in contraction force while slow-twitch muscles demonstrate prolonged half-relaxation time. However, such alterations have only been examined after a relatively short period following diabetes onset, with no information available regarding muscle damage caused by longer disease periods (>20 weeks). This study examined alterations in the contractile properties of the medial gastrocnemius (fast-twitch) and soleus (slow-twitch) muscles, as well as morphological changes in their motor neurons 12 and 22 weeks after diabetes onset. Adult male Wistar rats were divided into diabetic (12- or 22-week post-streptozotocin injection) and age-matched control groups. Electrically evoked maximum twitch and tetanic tension were recorded from leg muscles. Additionally, motor neuron number and cell body size were examined. At 12 weeks after diabetes onset, decreases in twitch force were observed predominantly in medial gastrocnemius muscles, while soleus muscles exhibited prolonged half-relaxation time. However, these differences became ambiguous at 22 weeks, with decreased twitch force and prolonged half-relaxation time observed in both muscles. On the other hand, reduction in soleus motor neurons was observed 12 weeks after diabetes onset, while medial gastrocnemius motor neurons were diminished at 22 weeks. These data indicate that experimental diabetes induces differential damage to medial gastrocnemius and soleus muscles as well as motor neurons. These diabetes-induced differences may partly underlie the differential deficits observed in gastrocnemius and soleus.

Effect of streptozotocin-induced diabetes on motor representations in the motor cortex and corticospinal tract in rats.

Motor disorders in patients with diabetes are associated with diabetic peripheral neuropathy, which can lead to symptoms such as lower extremity weakness. However, it is unclear whether central motor system disorders can disrupt motor function in patients with diabetes. In a streptozotocin-induced rat model of type 1 diabetes, we used intracortical microstimulation to evaluate motor representations in the motor cortex, recorded antidromic motor cortex responses to spinal cord stimulation to evaluate the function of corticospinal tract (CST) axons, and used retrograde labeling to evaluate morphological alterations of CST neurons. The diabetic rats exhibited size reductions in the hindlimb area at 4 weeks and in trunk and forelimb areas after 13 weeks, with the hindlimb and trunk area reductions being the most severe. Other areas were unaffected. Additionally, we observed reduced antidromic responses in CST neurons with axons projecting to lumbar spinal segments (CST-L) but not in those with axons projecting to cervical segments (CST-C). This was consistent with the observation that retrograde-labeled CST-L neurons were decreased in number following tracer injection into the spinal cord in diabetic animals but that CST-C neurons were preserved. These results show that diabetes disrupts the CST system components controlling hindlimb and trunk movement. This disruption may contribute to lower extremity weakness in patients.

Effect of streptozotocin-induced diabetes on motoneurons and muscle spindles in rats.

This study examined the alterations in the number and size of motoneurons innervating the medial gastrocnemius (MG) and biceps femoris (BF) motor nuclei in diabetic rats (12 or 22 weeks after injection of streptozotocin) and age-matched controls using retrograde labeling technique. Additionally, morphological alterations of muscle spindles in BF and MG muscles were tested. Significantly fewer labeled MG motoneurons were found in 12- and 22-week diabetic rats as compared with age-matched control animals. In contrast, the number of BF motoneurons was preserved in each group. Compared to control animals, the ratio of larger motoneurons of MG and BF muscle were decreased at 12 weeks, and smaller MG motoneurons were drastically decreased at 22 weeks. Moreover, MG muscle spindle showed reduction of its number and increase of intrafusal muscle fibers; however, BF muscle spindles showed little or no difference from control animals. We conclude that there is an early loss of alpha motoneurons for both MG and BF muscles followed by a later loss of gamma motoneurons in MG muscle in diabetic animals. Moreover, loss of gamma motoneuron might induce atrophy of MG muscle spindles.

Midkine in repair of the injured nervous system.

UNLABELLED: Midkine (MK) is a growth factor with neurotrophic and neurite outgrowth activities. It was expressed in the peri-ischaemic area in the acute phase of cerebral infarction in rat brains. Astrocytes were the origin of MK in this occasion. MK has been assessed in terms of its effects on neural injury. The administration of MK into the lateral ventricle immediately prior to ischaemia prevented cell death in the hippocampal CA1 neurons degenerated by transient forebrain ischaemia in gerbils. MK administration was also beneficial in rats with neural injury, especially after kainic acid-induced seizures. Gene therapy with mouse MK cDNA using an adenovirus was effective in reducing the cerebral infarction volume and in increasing the number of neuronal precursor cells in the subventricular zone of the rat brain. MK mRNA and MK protein were found in spinal cord motor neurons of the anterior horn in both the acute phase of sciatic nerve injury and 3 weeks later. MK immunoreactivity was also found in the proximal side of a sciatic nerve-injured site in sciatic nerve axons. MK receptors were expressed in Schwann cells after injury, suggesting crosstalk between axons and Schwann cells. MK was also present in nerve terminals and influenced ACh receptor clustering during neuromuscular development in Xenopus. Thus, MK may also be involved in reinforcing and maintaining the synapse. All these findings indicate the therapeutic potential of MK for promoting repair of the nervous system after injury. LINKED ARTICLES: This article is part of a themed section on Midkine. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-4.

Midkine-deficient mice delayed degeneration and regeneration after skeletal muscle injury.

Midkine (MK), a heparin-binding growth factor, was previously found to be expressed in the rat myotube-forming stage. We investigated MK gene-deficient (Mdk(-/-)) mice in terms of skeletal muscle degeneration and regeneration after injury by bupivacaine injection into the tibialis anterior muscle. Injured muscles showed intense inflammatory cell infiltration. Myotubes, myofibers with centrally located nuclei in their cytoplasm, were significantly smaller in Mdk(-/-) mice than in wild type (Mdk(+/+)) mice 7 days after injury (p=0.02). The distribution of myotube sizes showed quantitative differences between the two groups at 5 and 7 days, but not at 14 days. Many small myotubes were found in the regenerative area of Mdk(-/-) mice compared with that of Mdk(+/+)mice 5 and 7 days after injury. The expression of Iba1, a macrophage marker, was significantly lower in Mdk(-/-) mice 3 days after injury (p=0.01). The number of desmin-positive cells like myoblasts in Mdk(-/-) mice was significantly fewer than that in Mdk(+/+) mice 3 days after injury. Our results suggested that deletion of MK results in a delay in regeneration, preceded by decelerated migration of macrophages to the damaged area, and that MK has a role in cell differentiation and maturation after skeletal muscle injury.

Disruption of the midkine gene (Mdk) delays degeneration and regeneration in injured peripheral nerve.

Midkine (MK) is a growth factor implicated in the development and repair of various tissues, especially neural tissues. MK acts as a reparative neurotrophic factor in damaged peripheral nerves. A postulated role of MK in the degeneration and regeneration of sciatic nerves was explored by comparing wild-type (Mdk(+/+)) mice with MK-deficient (Mdk(-/-)) mice after freezing injury. In the Mdk(-/-) mice, a regenerative delay was observed, preceded by a decelerated Wallerian degeneration (WD). The relative wet weight of the soleus muscle slowly declined, and recovery was delayed compared with that in the Mdk(+/+) mice. In the regenerating nerve, unmyelinated axons were unevenly distributed, and some axons contained myelin-like, concentrically lamellated bodies. In the endplates of soleus muscles, nerve terminals containing synaptic vesicles disappeared in both mice. In Mdk(-/-) mice, the appearance of nerve terminals was delayed in synaptic vesicles of terminal buttons after injury. The recovery of evoked electromyogram was delayed in Mdk(-/-) mice compared with Mdk(+/+) mice. Our results suggested a delay in axonal degeneration and regeneration in Mdk(-/-) mice compared with Mdk(+/+) mice, and the delayed regeneration was associated with a delayed recovery of motor function. These findings show that a lack of MK following peripheral nerve injury is a critical factor in degeneration and regeneration, and manipulation of the supply of MK may offer interesting therapeutic options for the treatment of peripheral nerve damage.