Recovery of damaged skeletal muscle in mdx mice through low-intensity endurance exercise.

The lack of dystrophin in mdx mice leads to cycles of muscle degeneration and regeneration processes. Various strategies have been proposed in order to reduce the muscle-wasting component of muscular dystrophy, including implementation of an exercise programme. The aim of this study was to examine how low-intensity endurance exercise affects the degeneration-regeneration process in dystrophic muscle of male mdx mice. Mice were subjected to low-intensity endurance exercise by running on a motorized Rota-Rod for 5 days/week for 6 weeks. Histomorphological analysis showed a significant reduction of measured inflammatory-necrotic areas in both gastrocnemius and quadriceps muscle of exercised mdx mice as compared to matched sedentary mdx mice. The degenerative-regenerative process was also evaluated by examining the protein levels of connexin 39 (Cx39), a specific gene expressed in injured muscles. Cx39 was not detected in sedentary wild type mice, whereas it was found markedly increased in sedentary mdx mice, revealing active muscle degeneration-regeneration process. These Cx39 protein levels were significantly reduced in muscles of mdx mice exercised for 30 and 40 days, revealing together with histomorphological analysis a strong reduction of degeneration process in mice subjected to low-intensity endurance exercise. Muscles of exercised mdx mice did not show significant changes in force and fatigue resistance as compared to sedentary mdx mice. Overall in this study we found that specific low-intensity endurance exercise induces a beneficial effect probably by reducing the degeneration of dystrophic muscle.

Therapeutic potential of carbon monoxide in multiple sclerosis.

Carbon monoxide (CO) is produced during the catabolism of free haem, catalyzed by haem oxygenase (HO) enzymes, and its physiological roles include vasodilation, neurotransmission, inhibition of platelet aggregation and anti-proliferative effects on smooth muscle. In vivo preclinical studies have shown that exogenously administered quantities of CO may represent an effective treatment for conditions characterized by a dysregulated immune response. The carbon monoxide-releasing molecules (CORMs) represent a group of compounds capable of carrying and liberating controlled quantities of CO in the cellular systems. This review covers the physiological and anti-inflammatory properties of the HO/CO pathway in the central nervous system. It also discusses the effects of CORMs in preclinical models of inflammation. The accumulating data discussed herein support the possibility that CORMs may represent a novel class of drugs with disease-modifying properties in multiple sclerosis.

Creatine ingestion effects on oxidative stress in a steady-state test at 75% VO(2max).

AIM: The present study was carried out with the aim to analyze the role of creatine on oxidative stress during exercise, i.e. whether creatine is a pro-oxidative or an antioxidant substance. METHODS: In a randomized double-blind study involving 30 adult males, we examined plasma lactate, oxidative stress markers (malondialdehyde, MDA) and glutathione redox ratio (GSSG·GSH-1), antioxidative systems (vitamins A, E, C), and ergospirometric responses (respiratory quotient and relative oxygen uptake) before and after 30 min steady-state tests 75% VO2max (placebo and creatine). RESULTS: Ergospirometric tests, hematocrit values, blood lactate as well as vitamins A, E and C concentrations did not show significant differences between creatine and placebo testing. Conversely, oxidative stress markers MDA and GSSG·GSH-1 increased during placebo trials much more than in creatine trials. CONCLUSION: This is the first report documenting that a creatine loading, i.e., a 0.3 g/kg/die of creatine ingestion for 5 consecutive days, could reduce the oxidative stress, whereas its consumption may not have a clear metabolic advantage in certain aerobic activities.

Comparison of neuronal activities of external cuneate nucleus, spinocerebellar cortex and interpositus nucleus during passive movements of the rat's forelimb.

In this paper we examined the neuronal activities of external cuneate nucleus, spinocerebellar Purkinje cells and interpositus nucleus during passive forelimb movements in anesthetized rats with the aim of identifying common or different patterns of activation across structures. By means of principal components analysis, we identified two main patterns of discharge which explained most of the dataset variance. One component characterized the movement-related activity of external cuneate and spinocerebellar cortical neurons, while the other reflected neuronal activity of the interpositus nucleus. We also found that both principal components were related to global forelimb kinematics but, while most of the variance of the activity of external cuneate cells and spinocerebellar Purkinje cells was explained by the limb axis orientation and orientation velocity, interpositus neurons' firing was best related to length and length velocity. This difference in the forelimb kinematics representation observed in external cuneate nucleus and spinocerebellar cortex compared with the interpositus nucleus is discussed with respect to the specific role that these structures may play also during active control of limb movements.

Spatial anisotropy in the encoding of three-dimensional passive limb position by the spinocerebellum.

In an earlier study, we found that the encoding of limb position in the sagittal plane across the population of spinocerebellar Purkinje cells was anisotropic with a preferential gradient along horizontal direction. The aim of this study was to extend to a three-dimensional (3D) workspace the analysis of the relationships between Purkinje cells activity and rat's forelimb spatial position. In anesthetized animals, the extracellular activity of 121 neurons was recorded while a robot passively placed the limb in 18 positions within a cubic workspace (3x3x3 cm). In order to characterize the relationship between spatial locations and Purkinje cell activity we performed a backward stepwise regression starting from a model with three independent variables representing the antero-posterior, the medial-lateral and the vertical axes of workspace. Regression analysis showed that the firing of most cells was modulated exclusively along the antero-posterior (25%) or the medial-lateral (38%) axis, while a small portion was related only to the vertical axis (8%), indicating a generalized nonuniform sensitivity of Purkinje cells to limb displacement in 3D space.

Neuronal Encoding of Self and Others' Head Rotation in the Macaque Dorsal Prefrontal Cortex.

Following gaze is a crucial skill, in primates, for understanding where and at what others are looking, and often requires head rotation. The neural basis underlying head rotation are deemed to overlap with the parieto-frontal attention/gaze-shift network. Here, we show that a set of neurons in monkey's Brodmann area 9/46dr (BA 9/46dr), which is involved in orienting processes and joint attention, becomes active during self head rotation and that the activity of these neurons cannot be accounted for by saccade-related activity (head-rotation neurons). Another set of BA 9/46dr neurons encodes head rotation performed by an observed agent facing the monkey (visually triggered neurons). Among these latter neurons, almost half exhibit the intriguing property of encoding both execution and observation of head rotation (mirror-like neurons). Finally, by means of neuronal tracing techniques, we showed that BA 9/46dr takes part into two distinct networks: a dorso/mesial network, playing a role in spatial head/gaze orientation, and a ventrolateral network, likely involved in processing social stimuli and mirroring others' head. The overall results of this study provide a new, comprehensive picture of the role of BA 9/46dr in encoding self and others' head rotation, likely playing a role in head-following behaviors.

Blink reflex abnormalities in children with Tourette syndrome.

Tourette syndrome (TS) is a common disorder which typically occurs during childhood or early adolescence. There is no definitive diagnostic test for TS. The objective of this study was to demonstrate whether neurophysiological abnormalities of the blink reflex can be observed in children with TS. We enrolled 15 children with TS, diagnosed according to DSM IV Diagnostic Criteria, and 15 controls. The blink reflex was elicited by stimulating the supraorbital nerve in order to measure the early response (R1), homolateral and contralateral R2 (late) responses, amplitude of R1 and duration of R2. The mean duration of R2 was significantly longer in TS patients than in the controls (P < 0.001, Student's t-test). An abnormal pattern of the blink reflex can be, even in childhood, an early neurophysiologic marker of TS, which is not related to the duration of TS or to the age of onset.

Control of head stability during gait initiation in young and older women.

Transition tasks between static and dynamic situations may challenge head stabilization and balance in older individuals. The study was designed to investigate differences between young and older women in the upper body motion during the voluntary task of gait initiation. Seven young (25+/-2.3 years) and seven older healthy women (78+/-3.4 years) were required to stand on a force platform and initiate walking at their self-selected preferred speed. Angles of head, neck and trunk were measured by motion analysis in the sagittal plane and a cross-correlation analysis was performed on segments pairs. Variability of head and neck angular displacements, as indicated by average standard deviation, was significantly greater in the older than in the young participants. The young women maintained dynamic stability of the upper body, as forward flexion of the trunk was consistently counteracted by coordinated head-neck extension. Differently, movement patterns employed by the older women also included a rigid motion of all upper body segments leaning forward as a single unit. These results demonstrated that older women perform the transition from standing to walking with greater variability in the patterns of upper body motion compared to young women.

Organization of bilateral spinal projections to the lateral reticular nucleus of the rat.

The present study was carried out to analyze the topography of bilateral spinal projections to the lateral reticular nucleus (LRN). We used retrograde transport of fluorescent tracers Fast Blue and Diamidino Yellow to identify spinal neurons projecting to the ipsilateral and/or contralateral LRN, as well as orthograde transport of Phaseolus vulgaris leucoagglutinin to identify the LRN areas where spinoreticular axons terminate. Orthograde labeling confirmed that bilateral spinoreticular projections coming from cervical and upper-thoracic segments terminate in the magnocellular division of LRN, while those coming from the lower-thoracic, lumbar and sacral segments end in the parvocellular division of the nucleus; only a sparse spinal input has been observed in the subtrigeminal division of LRN. Retrograde labeling showed that labeled neurons were present at all spinal levels and in particular large numbers in the cervical and lumbar enlargements. Retrogradely single-labeled cells were located, with contralateral predominance, in all segments of the spinal cord, within laminae IV, V, VI, VIII, and X, whereas in laminae III and VII labeled neurons were mainly observed ipsilaterally. Furthermore, a small fraction of double-labeled cells (7.4%) was observed throughout the spinal cord, mainly in laminae III, IV, VII and VIII.

Orienting movements in area 9 identified by long-train ICMS.

The effect of intracortical microstimulation has been studied in several cortical areas from motor to sensory areas. The frontal pole has received particular attention, and several microstimulation studies have been conducted in the frontal eye field, supplementary eye field, and the premotor ear-eye field, but no microstimulation studies concerning area 9 are currently available in the literature. In the present study, to fill up this gap, electrical microstimulation was applied to area 9 in two macaque monkeys using long-train pulses of 500-700-800 and 1,000 ms, during two different experimental conditions: a spontaneous condition, while the animals were not actively fixating on a visual target, and during a visual fixation task. In these experiments, we identified backward ear movements, goal-directed eye movements, and the development of head forces. Kinematic parameters for ear and eye movements overlapped in the spontaneous condition, but they were different during the visual fixation task. In this condition, ear and eye kinematics have an opposite behavior: movement amplitude, duration, and maximal and mean velocities increase during a visual fixation task for the ear, while they decrease for the eye. Therefore, a top-down visual attention engagement could modify the kinematic parameters for these two effectors. Stimulation with the longest train durations, i.e., 800/1,000 ms, evokes not only the highest eye amplitude, but also a significant development of head forces. In this research article, we propose a new vision of the frontal oculomotor fields, speculating a role for area 9 in the control of goal-directed orienting behaviors and gaze shift control.

Origin of spinal projections to the anterior and posterior lobes of the rat cerebellum.

The present study was carried out to analyze the topography of spinal projections to the anterior and posterior lobes of the cerebellum and to investigate whether projections to the two lobes come from different spinocerebellar neurons or from branching axons of the same cells. We used orthograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) to identify the cerebellar areas where spinocerebellar axons terminate and retrograde double-labeling techniques to estimate the incidence of spinocerebellar neurons projecting to both anterior and posterior lobes via axon collaterals. Orthograde labeling confirmed that the rat, like other mammalian species, has spinocerebellar projections to two different regions of cerebellar cortex, i.e., lobules I-V of the anterior lobe and lobule VIII of the posterior lobe, with the highest incidence in lobules II, III, and VIII. We did not observe a clear difference in the distribution of afferents coming from different spinal segments to either of the two lobes. The double-labeled cells were located primarily in the lower thoracic and upper lumbar segments, almost exclusively in Clarke's column and in the dorso-lateral part of lamina 7 (in the region of the spinal border cells). It is likely that most or all of the spinocerebellar neurons in these structures project to both anterior and posterior lobes. Therefore, the two lobes of the cerebellum are likely to receive common information from these cells, but different information from the separate populations of spinocerebellar neurons that project only to one lobe or the other.

Cerebellar influences on accessory oculomotor nuclei of the rat: a neuroanatomical, immunohistochemical, and electrophysiological study.

With the aim to evaluate a possible neocerebellar control on eye movements, the projections from the cerebellar lateral nucleus (LN) to the accessory oculomotor nuclei (i.e., the nucleus of posterior commissure, the nucleus of Darkschewitsch, and the interstitial nucleus of Cajal), the putative neurotransmitters subserving this pathway, and the nature of the synaptic influences exerted by these projections were studied in adult rats. We used the orthograde transport of horseradish peroxidase conjugated with wheat germ agglutinin (WGA-HRP) to identify the mesencephalic areas where cerebellofugal fibers terminate, and retrograde labeling with the fluorescent dye fluoro-gold to estimate the incidence of cerebellar neurons projecting to the accessory oculomotor nuclei. Orthograde labeling showed that only a small contingent of cerebellofugal fibers reaches the contralateral accessory oculomotor nuclei. The retrogradely labeled cells were located primarily in the small-celled part of LN. By immunohistochemistry, we observed that all the cells retrogradely labeled from the accessory oculomotor area were also stained by using glutamate or aspartate antisera, but none of them were double-stained with a GABA antiserum. Electrical stimulation of the contralateral LN elicited changes in firing rate of a significant fraction of cells belonging to the accessory oculomotor nuclei (36.4% in the nucleus of posterior commissure, 47.1% in the nucleus of Darkschewitsch, and 44.6% in the interstitial nucleus of Cajal). In 57.8% of the cases, the responses were excitations, most of which had latencies and response characteristics compatible with a monosynaptic linkage. The remaining 42.2% of the cases were inhibitions with latencies ranging between 5 and 22 ms. Extracellular field potential recordings within the contralateral accessory oculomotor nuclei were interpreted as arising from impulses propagating along excitatory axons projecting in a bundle from the cerebellum. Stimulation of LN area in rats following intranuclear injection of kainic acid was not capable of evoking short latency excitations, so these responses can be considered to depend on the activation of LN efferents. The LN projection on accessory oculomotor nuclei could be part of the final precise control exerted by the neocerebellum on those brain structures concerned with movements of the eyes.

Substantia nigra influences on the reticulospinal neurons: an electrophysiological and ionophoretic study in cats and rats.

The influences exerted by the substantia nigra on reticulospinal neurons and the nature of the synaptic transmitter subserving these projections have been studied in adult cats and rats. Nigral stimulation evokes discharge changes in a significant number of reticulospinal cells (32.4% in cats and 39.1% in rats) on both ipsi- and contralateral sides. The responses were short-latency inhibitions (69.5% in cats and 76.5% in rats), short-latency excitations (22.2% in cats and 23.6% in rats) and in the remaining few cases long-latency excitations. Short-latency excitations, quite similar to the nigra-induced ones, were the predominant response pattern (95.2% in cats and 96.9% in rats) elicited on reticulospinal cells following stimulation of cerebral peduncle. The stimulation of substantia nigra in rats submitted to chronic ablation of sensorimotor cortex elicited only inhibitory responses while stimulation of nigral area in rats with intranigral injection of kainic acid was still capable of evoking short-latency excitations but not short-latency inhibitions. Therefore, the former can be ascribed to activation of corticoreticular fibers running in the cerebral peduncle whereas the latter can be considered as depending on activation of nigral efferents. The nigra-influenced cells were both 'fast' and 'slow' reticulospinal neurons and resulted mainly located in most rostral regions of reticular formation. Ionophoretic application of GABA suppressed the spontaneous firing of reticulospinal cells while no effect was observed following application of dopamine. The nigra-induced inhibitions were abolished by GABA-antagonist bicuculline and not by dopamine-antagonist fluphenazine.(ABSTRACT TRUNCATED AT 250 WORDS)

Projections from the intracerebellar nuclei to the ventral midbrain tegmentum in the rat.

The present study was undertaken to provide anatomical evidence, in the rat, for a direct projection from the cerebellum towards structures, other than the red nucleus, which belong to the ventral midbrain tegmentum, by using the retrograde as well as the anterograde horseradish peroxidase transport method. Following unilateral injection in the ventral midbrain tegmentum of horseradish peroxidase, free or conjugated to wheat germ agglutinin, sparing the red nucleus, retrogradely labeled neurons were found in the contralateral cerebellar lateral nucleus and, at lower density, in the interpositus nucleus. No labeled neurons were found in the fastigial nucleus of either side. Anterogradely labeled axons from lectin coupled horseradish peroxidase injection sites in the lateral and interpositus nuclei reached the contralateral ventral midbrain tegmentum. Terminal labeling was observed in the entire red nucleus as well as in the lateral division of the ventral tegmental area of Tsai, in the dorsal region of the substantia nigra pars compacta, and in the medial part of the retrorubral field. No terminal labeling was found in the caudal linear nucleus, interfascicular nucleus, peripeduncular nucleus, rostral linear nucleus of the raphe, substantia nigra pars lateralis and the substantia nigra pars reticulata. Terminal labeling was also not observed in the ventral midbrain tegmentum following horseradish peroxidase injection in lateral and interpositus nuclei of rats pretreated with kainic acid. In conclusion, it is noteworthy that, besides the red nucleus, the sole structures of ventral midbrain tegmentum receiving cerebellar efferents are those with a higher density of dopaminergic cells.

Interpositus nucleus influences on pyramidal tract neurons in the cat.

The influences of the interpositus nucleus on pyramidal tract neurons were investigated by stimulating, in unanesthetized cats, interpositus nucleus foci which activated single muscles in limbs, while recording unitary discharges of pyramidal tract neurons located in foci (area 4 gamma) from which contraction was obtained in the same muscles as those excited from interpositus nucleus (agonist pyramidal tract neurons), in their antagonist (antagonist pyramidal tract neurons), or in heteronymous muscles (heteronymous pyramidal tract neurons). It was found that agonist pyramidal tract neurons were inhibited from the interpositus nucleus, whereas antagonist pyramidal tract neurons displayed a pure excitatory or an excitatory-inhibitory pattern, and the heteronymous neurons were not significantly influenced. A direct activation of interposito-thalamic efferents could be responsible for these effects. In fact, unitary discharge changes of pyramidal tract neurons, elicited from interpositus nucleus stimulation, persisted after chronic intermediate cortex ablation and dentate nucleus lesions, and disappeared following coagulations in the ventrolateral nucleus of the thalamus. These results suggest that interpositus nucleus efferents, which activate a given muscle, via the rubrospinal pathway, could inhibit the discharge of pyramidal neurons controlling that muscle, via collaterals direct to the thalamic ventrolateral nucleus.

Pyramidal input to the intracerebellar nuclei of the cat.

In unanesthetized cats it has been found that pyramidal volleys elicited upon medullary pyramidal tract stimulation were capable of modifying the discharge of 41% of intracerebellar nuclear cells, via pontocerebellar systems impinging predominantly on the lateral cerebellar cortex. The incidence of responsive cells was 80% in the dentate nucleus compared with 10% in the fastigial nucleus, 11% in the anterior and 12% in the posterior division of the interpositus nucleus. The response was in 59% of the cases excitation followed by inhibition, in 30% of the cases a pure excitation and in 11% of the cases a pure inhibition. Excitation, pure or followed by inhibition, had a mean latency of 5.78 ms and a mean duration of 12.21 ms, while inhibition displayed a mean latency of 9.03 ms and a mean duration of 34.64 ms. The possible functional significance of the pyramidal input to the lateral cerebellum is briefly discussed in relation to a possible convergence of pyramidal and associative impulses in single cerebellar neurons.

Effects of cerebellar dentate nucleus GABAergic cells on rat inferior olivary neurons.

The present study was undertaken to analyze the effects on unitary activity of inferior olive (IO) neurons elicited by activation of cerebellar lateral nucleus (LN), in rats submitted to the chronic destruction of MDJ structures, i.e. in animals in which the LN-evoked effects in IO should be depended only on activation of GABAergic cells of LN. It has been observed that about two-thirds of the olivocerebellar neurons are significantly affected by LN stimulation, and > 68% of those cells were inhibited. Two-thirds of the inhibitory responses were compatible with a monosynaptic linkage, whereas the remaining inhibitions were probably due to polisynaptic linkages. The majority of LN-induced inhibitions was abolished or greatly reduced following application of GABA antagonists.

c-fos expression in the accessory oculomotor nuclei following neocerebellar stimulation.

The present study was carried out to determine whether, in the rat, the electric activation of the projection from the cerebellar lateral nucleus (LN) to the accessory oculomotor nuclei (AON; nucleus of posterior commissure, nucleus of Darkschewitsch, interstitial nucleus of Cajal) is capable of inducing c-fos expression. In particular, we compared the effects of a continuous LN stimulation at low-frequency (tonic stimulation) with those induced by high frequency pulse trains (phasic stimulation). The observed results show that the stimulation of LN induces c-fos expression in a significant proportion of neurones in the contralateral AON. Phasic stimulation was slightly more effective than tonic stimulation in producing c-fos expression.

Effects of Gabapentin and Topiramate in primary rat astrocyte cultures.

We studied in vitro the effects of anticonvulsant drugs Gabapentin and Topiramate on the production of reactive oxygen species and nitric oxide (NO), the activity of glutamine synthetase (GS), and cell viability in primary cultures of rat cortical astrocytes which are intimately involved in the normal functioning of neurons. We investigated the effects of these drugs at concentrations within the therapeutic range (1, 10 and 50 microg/ml). We observed that, in cultured astrocytes, Gabapentin induced a weak increase in the biosynthesis of NO, a mild decrease in GS activity and cell viability, and minor induction of a stress condition. Topiramate was observed to induce even greater stressor effects on these cells.

Cortical control of cerebellar dentato-rubral and dentato-olivary neurons.

The cortical input of 117 dentate nucleus neurons projecting either to the red nucleus (73 cells) or to the inferior olive (44 units) was studied electrophysiologically in rats. The majority of cells in both groups responded to electrical stimulation of discrete sites of the contralateral motor cortex. However, activation latencies from the same cortical focus were shorter for neurons projecting to the red nucleus than for olivary-projecting neurons. Principal components analysis pointed out significant differences between the two neuronal subgroups also in the temporal pattern of activity. These results suggest that a motor command might be transmitted through parallel independent channels to cerebellar neurons projecting to different regions of the brainstem.