Simone Marcuzzo

Treadmill training restores spatial cognitive deficits and neurochemical alterations in the hippocampus of rats submitted to an intracerebroventricular administration of streptozotocin

The intracerebroventricular infusion of streptozotocin (icv-STZ) has been largely used in research to mimic the main characteristics of Alzheimer’s disease (AD), including cognitive decline, impairment of cholinergic transmission, oxidative stress and astrogliosis. Moderate physical exercise has a number of beneficial effects on the central nervous system, as demonstrated both in animals and in human studies. This study aimed to evaluate the effect of 5-week treadmill training, in the icv-SZT model of sporadic AD, on cognitive function, oxidative stress (particularly mediated by NO) and on the astrocyte marker proteins, glial fibrillary acidic protein (GFAP) and S100B. Results confirm the spatial cognitive deficit and oxidative stress in this model, as well as astroglial alterations, particularly a decrease in CSF S100B. Physical exercise prevented these alterations, as well as increasing the hippocampal content of glutathione and GFAP per se in the CA1 region. These findings reinforce the potential neuroprotective role of moderate physical exercise. Astroglial changes observed in this dementia model contribute to understanding AD and other diseases that are accompanied by cognitive deficit.

Exercise improves motor deficits and alters striatal GFAP expression in a 6-OHDA-induced rat model of Parkinson’s disease

Astrocytic changes have been demonstrated in several neurodegenerative diseases, showing that these cells play an important role in functional recovery/maintenance against brain damage. Physical exercise is known to contribute to this process; however, the cellular mechanisms involved are not fully understood. This study investigated the effects of physical exercise on motor deficits and the expression of glial fibrillary acidic protein (GFAP) in a model of Parkinson’s disease (PD). Rats were divided into four groups: sham sedentary (SS) and sham trained (ST); lesioned sedentary (LS) and lesioned trained (LT). 6-OHDA was infused unilaterally into the medial forebrain bundle. Behavioral tasks were applied to evaluate motor abilities. Tyrosine hydroxylase (TH—in substantia nigra) and GFAP (in striatum) immunoreactivities (ir) were semi-quantified using optical density. The animals submitted to treadmill training completed fewer pharmacological-induced rotations when compared with sedentary animals and they also showed ameliorated motor impairments. Interestingly, although no change in TH-ir, the exercise led to restored striatal GFAP expression in the LT group while there was no effect in the ST group. This study is the first study to show data indicating the recovery of GFAP expression post-exercise in this model and further research is necessary to determine the precise action mechanisms of exercise on astrocytes in the PD.

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.

Dendritic spines in the posterodorsal medial amygdala after restraint stress and ageing in rats

Several evidences suggest that the posterodorsal medial amygdala (MePD) can be a relevant part of the rat neural circuitry for the regulation of hypothalamic neuroendocrine secretion and for ontogenetically different behavioral displays. The dendritic spine density of Golgi-impregnated neurons from the MePD was evaluated in young rats following acute or chronic restraint stress and in aged animals (24 months old). Compared to the control group, a single 1 h restraint stress session promoted a decreased spine density (p<0.01) whereas a single 6 h restraint stress session or daily 6-h restraint sessions for 28 consecutive days did not lead to the same effect (p>0.05). Aged rats showed no difference in this dendritic spine parameter when compared to young adults (p>0.05). These results indicate that short-term stress (1 h) can affect MePD dendritic spines and that neural plasticity is involved with adaptive responses onwards in restrained rats. On the other hand, brain structural modifications related with ageing appear not to influence the number of certain postsynaptic sites in the MePD of rats.

Effect of Treadmill Exercise on Serotonin Immunoreactivity in Medullary Raphe Nuclei and Spinal Cord Following Sciatic Nerve Transection in Rats

The serotoninergic system modulates nociceptive and locomotor spinal cord circuits. Exercise improves motor function and changes dopaminergic, noradrenergic, and serotonergic central systems. However, the direct relationship between serotonin, peripheral nerve lesion and aerobic treadmill exercise has not been studied. Using immunohistochemistry and optic densitometry, this study showed that the sciatic nerve transection increased the serotoninergic immunoreactivity in neuronal cytoplasm of the magnus raphe nuclei of trained and sedentary rats. In the dorsal raphe nucleus the increase only occurred in sedentary-sham-operated rats. In the spinal cord of trained, transected rats, the ventral horn showed significant changes, while the change in dorsal horn was insignificant. Von Frey’s test indicated analgesia in all exercise-trained rats. The sciatic nerve functional index indicated recovery in the trained group. Thus, both the aerobic treadmill exercise training and the nervous lesion appear to contribute to changes in serotonin immunoreactivity.

Enriched environment prevents memory deficits in type 1 diabetic rats.

Studies have shown that an enriched environmental (EE) enhances hippocampal neurogenesis and dendritic branching in rodents, improving the performance in learning and memory task. Diabetes, however, is associated with memory deficits and decreasing in cell proliferation in the hippocampal dentate gyrus (DG), possibly related with higher glucocorticoid levels. Thus, our objective was to investigate the influence of EE on the memory deficits and cell proliferation of diabetic rats. For this, we reared rats for 2 months during early stages of life in standard environments (control rats) or EE. At adulthood, control and EE groups were divided and half of them induced to diabetes by a single injection of streptozotocin, 60 mg/kg, via i.p. Memory deficit was evaluated in these groups in the novel object-placement recognition task 11 days after diabetes induction. BrdU label cells were detected by immunohistochemistry after 3 days of administration to correlate cell proliferation in the DG area and performance in the memory task. Our results showed that EE decreased memory deficits in diabetic-induced rats (p < 0.05). Although cell proliferation in the DG was lower in the diabetic rats, enriched environment did not interfere in this parameter. These findings suggest that enriched environment is able to prevent or delay the development of memory deficits caused by diabetes in rats.

Inflammatory response and oxidative stress in developing rat brain and its consequences on motor behavior following maternal administration of LPS and perinatal anoxia.

Cerebral palsy (CP) is a disorder of locomotion, posture and movement that can be caused by prenatal, perinatal or postnatal insults during brain development. An increased incidence of CP has been correlated to perinatal asphyxia and maternal infections during gestation. The effects of maternal exposure to low doses of bacterial endotoxin (lipopolysaccharide, LPS) associated or not with perinatal anoxia (PA) in oxidative and inflammatory parameters were examined in cerebral cortices of newborns pups. Concentrations of TNF‐α, IL‐1, IL‐4, SOD, CAT and DCF were measured by the ELISA method. Other newborn rats were assessed for neonatal developmental milestones from day 1 to 21. Motor behavior was also tested at P29 using open‐field and Rotarod. PA alone only increased IL‐1 expression in cerebral cortex with no changes in oxidative measures. PA also induced a slight impact on development and motor performance. LPS alone was not able to delay motor development but resulted in changes in motor activity and coordination with increased levels of IL‐1 and TNF‐α expression associated with a high production of free radicals and elevated SOD activity. When LPS and PA were combined, changes on inflammatory and oxidative stress parameters were greater. In addition, greater motor development and coordination impairments were observed. Prenatal exposure of pups to LPS appeared to sensitize the developing brain to effects of a subsequent anoxia insult resulting in an increased expression of pro‐inflammatory cytokines and increased free radical levels in the cerebral cortex. These outcomes suggest that oxidative and inflammatory parameters in the cerebral cortex are implicated in motor deficits following maternal infection and perinatal anoxia by acting in a synergistic manner during a critical period of development of the nervous system.

Olfactory and respiratory lamina propria transplantation after spinal cord transection in rats: Effects on functional recovery and axonal regeneration

Spinal cord injury (SCI) has very poor clinical prospects, resulting in irreversible loss of function below the injury site. Although applied in clinical trials, olfactory ensheathing cells transplantation (OEC) derived from lamina propria (OLP) is still a controversial repair strategy. The present study explored the efficacy of OLP or respiratory lamina propria (RLP) transplantation and the optimum period after SCI for application of this potential therapy. Adult male rats were submitted to spinal cord transection and underwent acute, 2-week or 4-week post-injury transplantation with pieces of OLP (containing OECs) or RLP (without OECs). After grafting, animals with OLP and RLP showed discrete and similar hindlimb motor improvement, with comparable spinal cord tissue sparing and sprouting in the lesion area. Acute transplantation of OLP and RLP seems to foster limited supraspinal axonal regeneration as shown by the presence of neurons stained by retrograde tracing in the brainstem nuclei. A larger number of 5-HT positive fibers were found in the cranial stump of the OLP and RLP groups compared to the lesion and caudal regions. Calcitonin gene-related peptide fibers were present in considerable numbers at the SCI site in both types of transplantation. Our results failed to verify differences between acute, 2-week and 4-week delayed transplantation of OLP and RLP, suggesting that the limited functional and axon reparative effects observed could not be exclusively related to OECs. A greater understanding of the effects of these tissue grafts is necessary to strengthen the rationale for application of this treatment in humans.

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.

Effects of fetal exposure to lipopolysaccharide, perinatal anoxia and sensorimotor restriction on motor skills and musculoskeletal tissue: implications for an animal model of cerebral palsy.

Cerebral palsy (CP) is a disorder of locomotion, posture and movement that can be caused by prenatal, perinatal or postnatal insults during brain development. An increased incidence of CP has been correlated to perinatal asphyxia and maternal infections during gestation. The effects of maternal exposure to low doses of bacterial endotoxin (lipopolysaccharide, LPS) on motor behavior and hind leg muscle morphology were examined in young adult rats. Prenatal exposure to LPS was also studied in association with perinatal anoxia (PA) and/or combined with subsequent sensorimotor restriction (SR) and all possible combinations of the three conditions. Rats exposed to LPS, PA and SR alone or combined (LPS + PA, LPS + SR, PA + SR, and LPS + PA + SR) showed deficits in balance and coordination when tested on the Rotarod. The SR groups, with or without other insults, (SR, LPS + SR, PA + SR, and LPS + PA + SR) exhibited the greatest motor deficits, characterized by the reduced ability to perform the horizontal ladder and suspended bar tests on postnatal day 29 (P29) and P45. Histological assessment revealed substantial morphological alterations in the slow ankle extensor soleus muscle of all SR rats. Soleus myofibers presented a reduction in cross-sectional area (CSA), an increase in sarcomere length and a decrease in sarcomere density. The CSA of the fast flexor tibialis anterior muscle was only decreased by the association of all treatments (LPS, PA, SR), but no differences were found in sarcomere length and density when compared to control. A slow-to-fast fiber type transition was only observed in the soleus and tibialis anterior muscles in the SR groups. These results suggest that exposure to LPS during the prenatal period, PA, SR alone or in combination has various degrees of consequences on motor behavior and muscle morphology. These data corroborate the concept that early experience-dependent movements play the most important role in shaping motor behavior and that reduced or anomalous sensorimotor experience can contribute to the development of aberrant motor behavior and muscle morphology.