H Majczynski

Different patterns of fore-hindlimb coordination during overground locomotion in cats with ventral and lateral spinal lesions.

The effect of large, low thoracic (T10–T11), partial spinal lesions involving the ventral quadrants of the spinal cord and, to a different extent, the dorsolateral funiculi, on fore-hindlimb coordination was examined in cats walking overground at moderate speeds (40–100 cm/s). Three different forms of impairment of fore-hindlimb coordination depending on the extent of the lesions, were observed. Lesions sparing the dorsolateral or the ventral funiculus on one side preserved the equality of the fore- and hindlimb locomotor rhythms but changed the coupling between the movements of both girdles as compared to intact animals. Larger lesions in which, in addition to the ventral quadrants of the spinal cord, also major parts of the dorsolateral funiculi were destroyed elicited episodes of rhythm oscillations in both girdles, which appeared at the background of a small difference in these rhythms. Lesions destroying almost the whole spinal cord induced a permanent difference (about 200 ms) in the step cycle duration of the fore- and the hindlimbs. However, even in these animals some remnant form of fore-hindlimb coordination was found. The results suggest that dorsolateral funiculi play a major role in preserving the equality of rhythms in the foreand the hindlimbs, while lesions of the ventral quadrants change the coupling between limbs.

Unrestrained walking in cats with partial spinal lesions.

In four cats with partial spinal lesions, performed at a low thoracic level, involving ventral quadrants and, to a different extent, the dorsolateral funiculi, several parameters of locomotion were analyzed during unrestrained walking at moderate speed (0.3-1.0 m/s). Special attention was paid to the analysis of support patterns and the durations of support phases in step cycles. The operated subjects displayed a much greater variability of support patterns than intact cats as well as changes in the relative duration of some support phases. The most striking difference was an increase in the relative duration of support on two homolateral limbs accompanied by a reduction of support on diagonal limbs. These changes were mainly due to an impairment of fore-hindlimb coordination as shown by an increase in the phase shifts between the movements of diagonal limbs. Other parameters of locomotion were essentially unaltered, except for cats in which the lesion destroyed bilaterally major portions of the dorsolateral funiculi.

EMG activity in response to static and dynamic facial expressions

The EMG activity associated with static and dynamic facial expressions (morphs with happy or angry emotions) were compared. We hypothesized that dynamic faces would (a) enhance facial muscular reactions and (b) evoke higher intensity ratings. Our analysis showed that dynamic expressions were rated as more intense than static ones. Subjects reacted spontaneously and rapidly to happy faces with increased zygomaticus major EMG activity and decrease corrugator supercilii EMG activity - showing greater changes in response to dynamic than to static stimuli in both muscles. In contrast, angry faces evoked no alteration of EMG activity in zygomaticus muscles and only small changes in the corrugator muscle EMG, and there was no difference between the responses to static and dynamic stimuli. It may be concluded that the dynamic property facilitates processing of facial expressions of emotions.

The upright posture improves plantar stepping and alters responses to serotonergic drugs in spinal rats

•  Locomotor training of rats held in an upright posture has been used recently to restore locomotion after spinal cord injury. Our results show that the upright posture alone improves locomotor recovery in spinal rats. •  This improvement is reversed by the removal of cutaneous afferent feedback from the paw, showing that sensory feedback from the foot facilitates the spinal central pattern generator (CPG) for locomotion. •  5‐HT2 and 5‐HT1A/7 agonists improve locomotion in the horizontal posture but can impair locomotion in the upright posture, suggesting that a proper balance of afferent feedback from the foot and 5‐HT receptor activation is necessary for optimal locomotor recovery. •  Our results provide new insights into the organization of the CPG for locomotion and the evolution of hominid bipedalism. The potent effects of cutaneous afferents from the paw revealed here must be taken into account in the design of strategies to restore locomotion after spinal cord injury.

Grafting of fetal brainstem 5-HT neurons into the sublesional spinal cord of paraplegic rats restores coordinated hindlimb locomotion

In rodent models of spinal cord injury, there is increasing evidence that activation of the locomotor central pattern generator (CPG) below the site of injury with 5-hydroxytryptamine (5-HT) agonists improves locomotor recovery and restores coordination. A promising means of replacing 5-HT control of locomotion is to graft brainstem 5-HT neurons into the spinal cord below the level of the spinal cord injury. However, it is not known whether this approach improves limb coordination because recovery of coordinated stepping has not been documented in detail in previous studies employing this transplantation strategy. Here, adult rats with complete spinal cord transections at the T9/10 level were grafted with E14 fetal neurons from the medulla at the T10/11 vertebra level one month after injury. The B1, B2 and B3 fetal anlagen of brainstem 5-HT neurons, a grouping that included the presumed precursors of recently described 5-HT locomotor command neurons, were used in these grafts. EMG and video recordings of treadmill locomotion evoked by tail stimulation showed full recovery of inter- and intralimb coordination in the grafted rats. We showed, using systemically applied antagonists, that 5-HT₂ and 5-HT₇ receptors mediate the improved locomotion after grafting, but through actions on different populations of spinal locomotor neurons. Specifically, 5-HT₂ receptors control CPG activation as well as motoneuron output, while 5-HT₇ receptors contribute primarily to activity of the locomotor CPG. These results are consistent with the roles for these receptors during locomotion in intact rodents and in rodent brainstem-spinal cord in vitro preparations.

CD44 is expressed in non-myelinating Schwann cells of the adult rat, and may play a role in neurodegeneration-induced glial plasticity at the neuromuscular junction

CD44 is a multifunctional cell surface glycoprotein which regulates cell-cell and cell-matrix interactions in a variety of tissues. In particular, the protein was found to be expressed in glial cells of developing, but not adult, peripheral nerves, where it takes part in signaling mediated by ErbB class of receptors for neuregulins. Here, we demonstrate, using high resolution morphological methods, tissue fractionation and RT-PCR, that CD44 is strongly expressed in terminal Schwann cell (TSC) at the neuromuscular junction (NMJ) of the adult rat skeletal muscle. As CD44 is also expressed by Schwann cells of the non-myelinated Remak bundles of the proximal peripheral nerves, it appears to be a marker of non-myelinating Schwann cell subpopulation. The analysis of transgenic rats bearing a mutated superoxide-dismutase gene (SOD1(G93A)) causing familial amyotrophic lateral sclerosis (ALS) revealed that TSC activation and morphological plasticity at the NMJ, caused by ongoing denervation-reinnervation is associated with a strong increase in CD44 expression therein. Notably, CD44 immunoreactivity is present in fine axon-escheating processes of the glial cells that guide reinnervation. In addition, we found that both in normal and SOD1(G93A) muscle, CD44 expressed in TSC partially colocalizes with immunoreactivities of neuregulin receptors ErbB2 and ErbB3. The colocalization appears to reflect a physical interaction, as evidenced by co-immunoprecipitation and fluorescence resonance energy transfer (FRET) analysis between CD44 and ErbB3. Importantly, TSC activation upon ALS-like neurodegeneration results in significant increase in molecular proximity of CD44 and ErbB3, which may have an impact on glial plasticity at the NMJ.

Unrestrained walking in intact cats

In five freely moving cats walking with speeds of 0.4-1.0 m/s several parameters of locomotion were investigated. Special attention was paid to the analysis of support patterns and the duration of support phases. The animals used almost exclusively (in 88 to 99% of steps) the 3-2-3-2-3-2-3-2 support pattern in which phases of support on three limbs alternated with phases of support on two limbs, homolateral and diagonal. The relative duration of support phases showed a tendency to decrease with increased locomotor velocity, except for the supports on diagonal limbs which slightly increased. The mean duration of the majority of support phases was similar and ranged between 12.2 and 14.5% of the step cycle. Phases of support on both hind- and one forelimb were somewhat (about 5%) shorter. It is concluded that the relative stability of support patterns and of the duration of support phases during walking observed in the present experiment may serve as a template for comparing changes in the gait produced by various CNS lesions.

Unrestrained walking in cats with medial pontine reticular lesions

The early postoperative effects of lesions, aimed to destroy the caudal pole of the nucleus reticularis pontis oralis (NRPO) and the rostral pole of the nucleus reticularis pontis caudalis (NRPC), were tested in freely moving cats, walking at moderate speed (0.4-1.0 m/s). In cats in which these structures were partly or completely destroyed, the main effect of lesions was an impairment of fore-hindlimb coordination, as shown by a change in the relationships between the lateral and diagonal time shift durations and the step cycle duration. In the second week after the surgery the values of the slopes of linear regressions relating these variables were markedly changed as compared to the preoperative data. The results suggest that the NRPO and NRPC are involved in maintaining the proper forehindlimb coordination during unrestrained locomotion in cats.

Comparison of two methods for quantitative assessment of unrestrained locomotion in the rat

Changes in locomotor movements induced by central and peripheral nerve injury or obtained as a result of pharmacological treatment are increasingly being investigated in rats. Several methods have been used to assess changes in the main locomotor indices, most of which are based on video recordings, usually with low time resolution, or on X-ray cinematographic recordings. Other methods are based on qualitative visual locomotor scoring systems like the BBB scale. We have analyzed locomotor indices in freely moving rats using two methods that can give quantitative results and which may be readily automated. One is based on detecting the onsets of swing and stance phases with contact electrodes (CE), while the second is based on recording the bursts of electromyographic activity (EMG) from the flexor and extensor muscles of each limb during the swing and stance phases, respectively. Besides the investigation of spontaneous locomotion in intact rats, our study also included an examination of locomotion on a ladder using EMG recording and analysis of locomotor disturbances following spinal cord hemisection, for which combined application of the two methods appeared to be useful. Overall, the EMG method appears to be more versatile than the CE method, although the use of both methods in parallel is recommended.

Changes in contractile properties of motor units of the rat medial gastrocnemius muscle after spinal cord transection

The effects of complete transection of the spinal cord at the level of Th9/10 on contractile properties of the motor units (MUs) in the rat medial gastrocnemius (MG) muscle were investigated. Our results indicate that 1 month after injury the contraction time (time‐to‐peak) and half‐relaxation time were prolonged and the maximal tetanic force in most of the MUs in the MG muscle of spinal rats was reduced. The resistance to fatigue also decreased in most of the MUs in the MG of spinal animals. Moreover, the post‐tetanic potentiation of twitches in MUs diminished after spinal cord transection. Criteria for the division of MUs into three types, namely slow (S), fast fatigue resistant (FR) and fast fatigable (FF), applied in intact animals, could not be directly used in spinal animals owing to changes in contractile properties of MUs. The ‘sag’ phenomenon observed in unfused tetani of fast units in intact animals essentially disappeared in spinal rats and it was only detected in few units, at low frequencies of stimulation only. Therefore, the MUs in spinal rats were classified as fast or slow on the basis of an adjusted borderline of 20 ms, instead of 18 ms as in intact animals, owing to a slightly longer contraction time of those fast motor units with the ‘sag’. We conclude that all basic contractile properties of rat motor units in the medial gastrocnemius muscle are significantly changed 1 month after complete spinal cord transection, with the majority of motor units being more fatigable and slower than those of intact rats.