Patrícia Severo do Nascimento

Beneficial effects of treadmill training in experimental diabetic nerve regeneration.

OBJECTIVES: We investigated the effects of treadmill training (10 weeks) on hindlimb motor function and nerve morphometric parameters in diabetic rats submitted to sciatic nerve crush. MATERIALS AND METHOD: Wistar rats (n = 64) were divided into the following groups: non-diabetic; trained non-diabetic; non-diabetic with sciatic nerve crush; trained non-diabetic with sciatic nerve crush; diabetic; trained diabetic; diabetic with sciatic nerve crush or trained diabetic with sciatic nerve crush. Diabetes was induced by streptozotocin injection (50 mg/kg, iv). Hindlimb motor function was evaluated weekly by assessing sciatic functional indices, and the proximal and distal portions of the sciatic nerve were used for morphometric analysis. RESULTS: At 13 weeks post-injury, the distal nerve portion of all injured groups and the proximal nerve portion of the diabetic with sciatic nerve crush group presented altered morphometric parameters such as decreased myelinated fiber diameter (∼7.4±0.3µm vs ∼4.8±0.2µm), axonal diameter (∼5±0.2µm vs ∼3.5±0.1µm) and myelin sheath thickness (∼1.2±0.07µm vs ∼0.65±0.07µm) and an increase in the percentage of area occupied by endoneurium (∼28±3% vs ∼60±3%). In addition, in the non-diabetic with sciatic nerve crush group the proximal nerve portion showed a decreased myelinated fiber diameter (7.4±0.3µm vs 5.8±0.3µm) and myelin sheath thickness (1.29±0.08µm vs 0.92±0.08µm). The non-diabetic with sciatic nerve crush, trained non-diabetic with sciatic nerve crush, diabetic with sciatic nerve crush and trained diabetic with sciatic nerve crush groups showed normal sciatic functional index from the 4th, 4th, 9th and 7th week post-injury, respectively. Morphometric alterations in the proximal nerve portion of the diabetic with sciatic nerve crush and non-diabetic with sciatic nerve crush groups were either prevented or reverted to values similar to the non-diabetic group by treadmill training. CONCLUSION: Diabetic condition promoted delay in sciatic nerve regeneration. Treadmill training is able to accelerate hindlimb motor function recovery in diabetic injured rats and prevent or revert morphometric alterations in proximal nerve portions in non-diabetic and diabetic injured rats.

Beneficial effects of treadmill training in a cerebral palsy-like rodent model: walking pattern and soleus quantitative histology.

The aim of the present study was to investigate whether treadmill locomotor training could have beneficial effects on deficits consequent to perinatal anoxia, sensorimotor restriction or a combination of both. Fifty-six newborn male Wistar rats were divided into four groups: control, anoxic, sensorimotor-restricted and anoxic-sensorimotor-restricted. Rats were exposed to anoxia in the first two postnatal days (P0 and P1) and/or hind-limb sensorimotor restriction from P2 to P28 for 16 h/day. Control and experimental rats underwent treadmill training for three weeks (from P31 to P52). Body weight and walking patterns (stride length and foot angle) were measured weekly during treadmill locomotor training. Soleus muscle cross-sectional mean area and fiber density were measured using planar morphometry. Anoxia per se did not cause gait or muscle deficits. Body weight, stride length and soleus fiber cross-sectional mean area, however, were increased in trained anoxic rats. Sensorimotor-restricted animals, either with or without perinatal anoxia, showed deficits in body weight gain, decreased stride length, wider foot angle and soleus fiber atrophy. In the sensorimotor-restricted group, treadmill training improved body weight gain and stride length, and decreased the percentage of the atrophic fibers. However, in the anoxic-sensorimotor-restricted group, training improved stride length only. Three weeks of treadmill training were able to improve stride length in restricted and anoxic-restricted animals, although body weight deficit and the degree of degradation in muscle histology were reduced only in the restricted group.

The Beneficial Effects of Treadmill Step Training on Activity- Dependent Synaptic and Cellular Plasticity Markers After Complete Spinal Cord Injury

Several studies have shown that treadmill training improves neurological outcomes and promotes plasticity in lumbar spinal cord of spinal animals. The morphological and biochemical mechanisms underlying these phenomena remain unclear. The purpose of this study was to provide evidence of activity-dependent plasticity in spinal cord segment (L5) below a complete spinal cord transection (SCT) at T8–9 in rats in which the lower spinal cord segments have been fully separated from supraspinal control and that subsequently underwent treadmill step training. Five days after SCT, spinal animals started a step-training program on a treadmill with partial body weight support and manual step help. Hindlimb movements were evaluated over time and scored on the basis of the open-field BBB scale and were significantly improved at post-injury weeks 8 and 10 in trained spinal animals. Treadmill training also showed normalization of withdrawal reflex in trained spinal animals, which was significantly different from the untrained animals at post-injury weeks 8 and 10. Additionally, compared to controls, spinal rats had alpha motoneuronal soma size atrophy and reduced synaptophysin protein expression and Na+, K+-ATPase activity in lumbar spinal cord. Step-trained rats had motoneuronal soma size, synaptophysin expression and Na+, K+-ATPase activity similar to control animals. These findings suggest that treadmill step training can promote activity-dependent neural plasticity in lumbar spinal cord, which may lead to neurological improvements without supraspinal descending control after complete spinal cord injury.

Treadmill training improves motor skills and increases tyrosine hydroxylase immunoreactivity in the substantia nigra pars compacta in diabetic rats.

The aim of this study was to evaluate the effects of treadmill training on motor skills and immunoreactivity to tyrosine hydroxylase in the substantia nigra pars compacta and ventral tegmental area from diabetic rats induced by streptozotocin. Male Wistar rats were divided into three groups: control, diabetic and trained diabetic. Treadmill training was performed for 8weeks. Blood glucose concentrations and body weight were evaluated 48h after diabetes induction and every 30days thereafter. Motor skills were evaluated on the rotarod and open field tests. Then, animals were transcardially perfused and the brains were post-fixed, cryoprotected and sectioned in a cryostat. Immunohistochemistry for tyrosine hydroxylase analyses was done in the ventral tegmental area and in the substantia nigra. Motor skills showed that diabetic animals had a decrease in the latency to fall and enhanced number of falls in the rotarod test compared to control and trained diabetic animals. In the open field, diabetic animals had a decrease in the number of crossed squares, rearings and spent a less time moving compared to control and trained diabetic animals. In diabetic animals, optical densitometry of immunohistochemistry showed that tyrosine hydroxylase reaction decreased in the ventral tegmental area and in the neurons and process in the substantia nigra. In the later region, that decrease was reversed by treadmill training. In conclusion, we demonstrated that treadmill training can reverse the loss of the motor skills, which was correlated to tyrosine hydroxylase immunoreactivity in the substantia nigra of diabetic animals without pharmacological treatment.

Different effects of anoxia and hind-limb immobilization on sensorimotor development and cell numbers in the somatosensory cortex in rats

Cerebral palsy (CP) is a group of movement and posture disorders attributed to insults in the developing brain. In rats, CP-like motor deficits can be induced by early hind-limb sensorimotor restriction (SR; from postnatal days P2 to P28), associated or otherwise with perinatal anoxia (PA; on P0 and P1). In this study, we address the question of whether PA, early SR or a combination of both produces alterations to sensorimotor development. Developmental milestones (surface righting, cliff aversion, stability on an inclined surface, proprioceptive placing, auditory startle, eye opening) were assessed daily from P3 to P14. Motor skills (horizontal ladder and beam walking) were evaluated weekly (from P31 to P52). In addition, on P52, the thickness of the somatosensory (S1) and cerebellar cortices, and corpus callosum were measured, and the neuronal and glial cell numbers in S1 were counted. SR (with or without PA) significantly delayed the stability on an inclined surface and hastened the appearance of the placing reflex and impaired motor skills. No significant differences were found in the thickness measurements between the groups. Quantitative histology of S1 showed that PA, either alone or associated with SR, increased the number of glial cells, while SR alone reduced neuronal cell numbers. Finally, the combination of PA and SR increased the size of neuronal somata. We conclude that SR impairs the achievement of developmental milestones and motor skills. Moreover, both SR and PA induce histological alterations in the S1 cortex, which may contribute to sensorimotor deficits.

Physical exercise down-regulated locomotor side effects induced by haloperidol treatment in Wistar rats

Extra-pyramidal symptoms (EPS) such as akinesia, dystonia, gait alteration and tremors are observed when dopamine D2-receptors are blocked by pharmacological agents such as haloperidol. These alterations produce a Parkinson disease-like state (PLS). Physical exercise has been proven to improve gait and locomotor symptoms in Parkinsons disease; we sought to elucidate the effects of physical exercise on PLS induced by chronic administration of haloperidol in rats. We used 48 rats distributed into four groups: Control, Exercise, Haloperidol, and Hal+Exe. All the animals received a daily injection of saline or haloperidol for 30 days, and the exercise groups underwent a daily 30-minute exercise protocol for 20 days. The animals were subjected to the ink-paw test, bar test and open-field test throughout the training period. The haloperidol-induced akinesia increased throughout the days of injections, but exercise was shown to alleviate it. The assessment showed shortened stride length and increased stance width with the use of haloperidol, which were significantly alleviated by exercise. These results indicate that exercise could be an interesting approach towards reducing unwanted EPS caused by haloperidol.

Treadmill training induces plasticity in spinal motoneurons and sciatic nerve after sensorimotor restriction during early postnatal period: new insights into the clinical approach for children with cerebral palsy.

The aim of the present study was to investigate whether locomotor stimulation training could have beneficial effects on the morphometric alterations of spinal cord and sciatic nerve consequent to sensorimotor restriction (SR). Male Wistar rats were exposed to SR from postnatal day 2 (P2) to P28. Control and experimental rats underwent locomotor stimulation training in a treadmill for three weeks (from P31 to P52). The cross‐sectional area (CSA) of spinal motoneurons innervating hind limb muscles was determined. Both fiber and axonal CSA of myelinated fibers were also assessed. The growth‐related increase in CSA of motoneurons in the SR group was less than controls. After SR, the mean motoneuron soma size was reduced with an increase in the proportion of motoneurons with a soma size of between 0 and 800 μm2. The changes in soma size of motoneurons were accompanied by a reduction in the mean fiber and axon CSA of sciatic nerve. The soma size of motoneurons was reestablished at the end of the training period reaching controls level. Our results suggest that SR during early postnatal life retards the growth‐related increase in the cell body size of motoneurons in spinal cord and the development of sciatic nerve. Additionally, three weeks of locomotor stimulation using a treadmill seems to have a beneficial effect on motoneurons’ soma size.

Treadmill step training-induced adaptive muscular plasticity in a chronic paraplegia model.

The purpose of this study was to provide evidence that treadmill step training is capable of attenuating muscle atrophy and may regulate brain derived neurotrophic factor (BDNF) in soleus muscle after complete spinal cord transection (SCT) at T8-T9 in rats. Five days after SCT, spinal animals started a 9-week step-training program on a treadmill with partial body weight support and manual step help. The muscular trophism was studied by analyzing muscle weight and myofiber cross-sectional area of the soleus, while Western blot analysis was used to detect BDNF expression in the same muscle. Step training, initiated immediately after SCT in rats, may partially impede/revert muscular atrophy in chronic paralyzed soleus muscle. Moreover, treadmill step training promoted upregulation of the BDNF in soleus muscle, which was positively correlated with muscle weight and myofiber cross-sectional size. These findings have important implications for the comprehension of the neurobiological substrate that promotes exercise-induced effects on paralyzed skeletal muscle and suggests treadmill training is a viable therapeutic approach in spinal cord injuries.

Diabetes increases mechanical sensitivity and causes morphological abnormalities in the sural nerve that are prevented by treadmill training

In this investigation we evaluated the effects of treadmill training on mechanical sensitivity and sural nerve morphology in diabetic rats.

Exercise training improves the soleus muscle morphology in experimental diabetic nerve regeneration

In this study we evaluate the effects of exercise training (10 weeks) on soleus muscle morphology in diabetic nerve regeneration after injury by sciatic nerve crush.