Motor learning changes the axon initial segment of the spinal motoneuron.

We are studying the mechanisms of H-reflex operant conditioning, a simple form of learning. Modelling studies in the literature and our previous data suggested that changes in the axon initial segment (AIS) might contribute. To explore this, we used blinded quantitative histological and immunohistochemical methods to study in adult rats the impact of H-reflex conditioning on the AIS of the spinal motoneuron that produces the reflex. Successful, but not unsuccessful, H-reflex up-conditioning was associated with greater AIS length and distance from soma; greater length correlated with greater H-reflex increase. Modelling studies in the literature suggest that these increases may increase motoneuron excitability, supporting the hypothesis that they may contribute to H-reflex increase. Up-conditioning did not affect AIS ankyrin G (AnkG) immunoreactivity (IR), p-p38 protein kinase IR, or GABAergic terminals. Successful, but not unsuccessful, H-reflex down-conditioning was associated with more GABAergic terminals on the AIS, weaker AnkG-IR, and stronger p-p38-IR. More GABAergic terminals and weaker AnkG-IR correlated with greater H-reflex decrease. These changes might potentially contribute to the positive shift in motoneuron firing threshold underlying H-reflex decrease; they are consistent with modelling suggesting that sodium channel change may be responsible. H-reflex down-conditioning did not affect AIS dimensions. This evidence that AIS plasticity is associated with and might contribute to H-reflex conditioning adds to evidence that motor learning involves both spinal and brain plasticity, and both neuronal and synaptic plasticity. AIS properties of spinal motoneurons are likely to reflect the combined influence of all the motor skills that share these motoneurons. KEY POINTS: Neuronal action potentials normally begin in the axon initial segment (AIS). AIS plasticity affects neuronal excitability in development and disease. Whether it does so in learning is unknown. Operant conditioning of a spinal reflex, a simple learning model, changes the rat spinal motoneuron AIS. Successful, but not unsuccessful, H-reflex up-conditioning is associated with greater AIS length and distance from soma. Successful, but not unsuccessful, down-conditioning is associated with more AIS GABAergic terminals, less ankyrin G, and more p-p38 protein kinase. The associations between AIS plasticity and successful H-reflex conditioning are consistent with those between AIS plasticity and functional changes in development and disease, and with those predicted by modelling studies in the literature. Motor learning changes neurons and synapses in spinal cord and brain. Because spinal motoneurons are the final common pathway for behaviour, their AIS properties probably reflect the combined impact of all the behaviours that use these motoneurons.

Estimates of persistent inward currents in lower limb muscles are not different between inactive, resistance-trained, and endurance-trained young males.

Persistent inward currents (PICs) increase the intrinsic excitability of α-motoneurons. The main objective of this study was to compare estimates of α-motoneuronal PICs between inactive, chronic resistance-trained, and chronic endurance-trained young individuals. We also aimed to investigate whether there is a relationship in the estimates of α-motoneuronal PIC magnitude between muscles. Estimates of PIC magnitude were obtained in three groups of young individuals: resistance-trained (n = 12), endurance-trained (n = 12), and inactive (n = 13). We recorded high-density surface electromyography (HDsEMG) signals from tibialis anterior (TA), gastrocnemius medialis (GM), soleus (SOL), vastus medialis (VM), and vastus lateralis (VL). Then, signals were decomposed with convolutive blind source separation to identify motor unit (MU) spike trains. Participants performed triangular isometric contractions to a peak of 20% of their maximum voluntary contraction. A paired-motor-unit analysis was used to calculate ΔF, which is assumed to be proportional to PIC magnitude. Despite the substantial differences in physical training experience between groups, we found no differences in ΔF, regardless of the muscle. Significant correlations of estimates of PIC magnitude were found between muscles of the same group (VL-VM, SOL-GM). Only two correlations (out of 8) between muscles of different groups were found (TA-GM and VL-GM). Overall, our findings suggest that estimates of PIC magnitude from lower-threshold MUs at low contraction intensities in the lower limb muscles are not influenced by physical training experience in healthy young individuals. They also suggest muscle-specific and muscle group-specific regulations of the estimates of PIC magnitude.NEW & NOTEWORTHY Chronic resistance and endurance training can lead to specific adaptations in motor unit activity. The contribution of α-motoneuronal persistent inward currents (PICs) to these adaptations is currently unknown in healthy young individuals. Therefore, we studied whether estimates of α-motoneuronal PIC magnitude are higher in chronically trained endurance- and resistance-trained individuals. We also studied whether there is a relationship between the estimates of α-motoneuronal PIC magnitude of different lower limb muscles.

Strength versus endurance trained master athletes: Contrasting neurophysiological adaptations.

Neural factors play a critical role in the age-related decline in maximal strength and rate of force development (RFD). However, it is uncertain how the age-related attenuation in neuromuscular function may be mitigated in strength or endurance trained master athletes. In this study we applied evoked spinal motoneuron recordings to examine descending motor drive, i.e., efferent drive from supraspinal and spinal centres during maximal voluntary contraction (MVC; V-wave) and H-reflex excitability measured at 10 % MVC in older (>65 yrs) and younger (<35 yrs) strength athletes (n = 21), endurance athletes (n = 17) and untrained control participants (n = 30). Both strength (b = 0.09 [0.01-0.18], p = 0.038) and endurance training (b = 0.14 [0.04-0.23], p = 0.006) were associated with a high V-wave amplitude. This was likely explained by an elevated H-reflex excitability (b = 0.23 [0.11-0.35], p < 0.001) in endurance trained participants, which failed to be seen in strength trained participants. These contrasting neurophysiological properties were accompanied by different physiological traits; strength training was associated with high maximal strength (b = 107.5 [84.6 to 130.4] kg, p < 0.001) and RFD (b = 3171 [2248 to 4094] N‧s-1, p < 0.001), whereas endurance training was associated with elevated maximal oxygen uptake (V̇O2max; b = 13.6 [8.0-19.2] ml‧kg-1‧min-1, p < 0.001). This pattern was apparent irrespective of age, although all traits were negatively associated with advanced age (p < 0.05). In conclusion, strength trained individuals demonstrate higher descending motor drive (elevated V-wave responses), compared to age-matched untrained individuals. Endurance trained individuals also showed elevated V-wave responses, uniquely accompanied by enhanced α-motoneuron excitability and/or reduced pre/postsynaptic inhibition (elevated H-reflex responses). Since a high descending motor drive is a key component of strong muscle contractions, strength training should be emphasized to sustain the ability to carry out force-dependent tasks at older age.

Culture of Human iPSC-Derived Motoneurons in Compartmentalized Microfluidic Devices and Quantitative Assays for Studying Axonal Phenotypes.

In order to use induced Pluripotent Stem Cells (iPSCs) to model neurodegenerative diseases, efficient and homogeneous generation of neurons in vitro represents a key step. Here we describe a method to obtain and characterize functional human spinal and cranial motoneurons using a combined approach of microfluidic chips and programs designed for scientific multidimensional imaging. We have used this approach to analyze axonal phenotypes. These tools are useful to investigate the cellular and molecular bases of neuromuscular diseases, including amyotrophic lateral sclerosis and spinal muscular atrophy.

Acupuncture alleviates spinal hyperreflexia and motor dysfunction in post-ischemic stroke rats with spastic hypertonia via KCC2-mediated spinal GABAA activation.

The majority of patients simultaneously develop motor dysfunction and spastic hypertonia after ischemic strokes, which can be associated with an increasing trend in motor impairments, seriously impeding the rehabilitation process. Evidence suggests that some deficits in the KCC2 expression in the spinal cord along with maladaptive endogenous plasticity via GABAA receptors are often involved in the pathology of spastic hypertonia after a stroke. In this respect, acupuncture has been commonly used in clinical settings for post-stroke patients' rehabilitation. Nevertheless, the mechanism of the modulating activity of this alternative medicine in the spinal pathways to relieve spasticity and improve functional recovery after a stroke has still remained unclear. Utilizing laser speckle imaging, functional assessments (viz. neurologic function scale, muscular tension scale, foot balance test, and gait analysis), H-reflex recording, TTC, Western blotting, RT-qPCR, ELISA, and immunofluorescence molecular assay, the study results illustrated that acupuncture could significantly alleviate the spinal hyperreflexia, decrease muscle tone, and enhance locomotor function by elevating the GABA, KCC2, and GABAAγ2 expressions in the lumbar spine of a rat model of post-ischemic stroke with spastic hypertonia. Furthermore, the KCC2 antagonist DIOA abolished the benefits induced by this practice. Overall, the data revealed that acupuncture is a promising therapeutic approach for spastic hypertonia after a stroke, and the positive outcomes in this sense could be achieved via activating the KCC2-mediated spinal GABAA signaling pathway.

M-wave and H-reflex recruitment curves in boys and men.

The aim of the present study was to check whether the M-wave and H-reflex recruitment curves differ between prepubertal boys and men. Eleven boys (9-11 yr) and eleven men (18-35 yr) were magnetically stimulated at the tibial nerve in a prone position. M-wave and H-reflex maximal amplitudes (Hmax; Mmax ; Hmax /Mmax ), thresholds, regression slopes (Hslp ; Mslp ; Hslp /Mslp ) were extracted from M-wave and H-reflex recruitment curves and compared between the two age groups. Overall, no significant difference in M-wave and H-reflex recruitment curve parameters was found between the two populations. Nevertheless, the size of the M-wave associated with maximal H-reflex amplitude was lower in boys as compared to men when expressed relative to maximal M-wave amplitude (MHmax /Mmax : 0.18 ± 0.06 vs. 0.31 ± 0.13; p < .05). This result suggests that the development of peripheral nerve was completed in 9 to 11-year-old boys and did not affect the M-wave and H-reflex recruitment curves parameters. In neuromuscular function studies, it implies that Hmax /Mmax and Hslp /Mslp could be used indifferently to compare spinal motoneuron excitability between 9-11-year-old boys and men. Conversely, evoking H-reflexes at a given percentage of Mmax may bias the comparison between boys and men.

Motoneuron expression profiling identifies an association between an axonal splice variant of HDGF-related protein 3 and peripheral myelination.

Disorders that disrupt myelin formation during development or in adulthood, such as multiple sclerosis and peripheral neuropathies, lead to severe pathologies, illustrating myelin's crucial role in normal neural functioning. However, although our understanding of glial biology is increasing, the signals that emanate from axons and regulate myelination remain largely unknown. To identify the core components of the myelination process, here we adopted a microarray analysis approach combined with laser-capture microdissection of spinal motoneurons during the myelinogenic phase of development. We identified neuronal genes whose expression was enriched during myelination and further investigated hepatoma-derived growth factor-related protein 3 (HRP3 or HDGFRP3). HRP3 was strongly expressed in the white matter fiber tracts of the peripheral (PNS) and central (CNS) nervous systems during myelination and remyelination in a cuprizone-induced demyelination model. The dynamic localization of HPR3 between axons and nuclei during myelination was consistent with its axonal localization during neuritogenesis. To study this phenomenon, we identified two splice variants encoded by the HRP3 gene: the canonical isoform HRP3-I and a newly recognized isoform, HRP3-II. HRP3-I remained solely in the nucleus, whereas HRP3-II displayed distinct axonal localization both before and during myelination. Interestingly, HRP3-II remained in the nuclei of unmyelinated neurons and glial cells, suggesting the existence of a molecular machinery that transfers it to and retains it in the axons of neurons fated for myelination. Overexpression of HRP3-II, but not of HRP3-I, increased Schwann cell numbers and myelination in PNS neuron-glia co-cultures. However, HRP3-II overexpression in CNS co-cultures did not alter myelination.

Spinal Motoneuron TMEM16F Acts at C-boutons to Modulate Motor Resistance and Contributes to ALS Pathogenesis.

Neuronal Ca2+ entry elicited by electrical activity contributes to information coding via activation of K+ and Cl- channels. While Ca2+-dependent K+ channels have been extensively studied, the molecular identity and role of Ca2+-activated Cl- channels (CaCCs) remain unclear. Here, we demonstrate that TMEM16F governs a Ca2+-activated Cl- conductance in spinal motoneurons. We show that TMEM16F is expressed in synaptic clusters facing pre-synaptic cholinergic C-boutons in α-motoneurons of the spinal cord. Mice with targeted exon deletion in Tmem16f display decreased motor performance under high-demanding tasks attributable to an increase in the recruitment threshold of fast α-motoneurons. Remarkably, loss of TMEM16F function in a mouse model of amyotrophic lateral sclerosis (ALS) significantly reduces expression of an activity-dependent early stress marker and muscle denervation, delays disease onset, and preserves muscular strength only in male ALS mice. Thus, TMEM16F controls motoneuron excitability and impacts motor resistance as well as motor deterioration in ALS.

Expression of aromatase and estrogen receptors in lumbar motoneurons of mice.

Estrogen exerts protective roles in amyotrophic lateral sclerosis (ALS). However, the expression of aromatase (ARO) and estrogen receptors (ERs) in the motoneurons of spinal cord, has not yet been elucidated. By immunohistochemistry, we found that ARO and ERs were present in the ventral horn of adult mice lumbar spinal cord, and colocalized with SMI-32, a motoneuron specific marker. Within motoneurons, we observed that ARO is detected primarily in the cytoplasm, with fewer ARO in the nucleus; ERα and ERß mainly localized in the nucleus with less in the cytoplasm; while GPR30 is located in soma and processes. In conclusion, we found that ERs and ARO are expressed in the motoneurons of lumbar spinal cord in adult mice. These findings suggest that estrogen may be useful as a promising therapeutic agent for prevention of damage and improvement of locomotor function in ALS.

Muscle length-dependent contribution of motoneuron Cav1.3 channels to force production in model slow motor unit.

Persistent inward current (PIC)-generating Cav1.3 channels in spinal motoneuron dendrites are thought to be actively recruited during normal behaviors. However, whether and how the activation of PIC channels influences force output of motor unit remains elusive. Here, building a physiologically realistic model of slow motor unit I demonstrated that force production induced by the PIC activation is much smaller for short than lengthened muscles during the regular firing of the motoneuron that transitions from the quiescent state by either a brief current pulse at the soma or a brief synaptic excitation at the dendrites. By contrast, the PIC-induced force potentiation was maximal for short muscles when the motoneuron switched from a stable low-frequency firing state to a stable high-frequency firing state by the current pulse at the soma. Under the synaptic excitation at the dendrites, however, the force could not be potentiated by the transitioning of the motoneuron from a low- to a high-frequency firing state due to the simultaneous onset of PIC at the dendrites and firing at the soma. The strong dependency of the input-output relationship of the motor unit on the neuromodulation and Ia afferent inputs for the PIC channels was further shown under static variations in muscle length. Taken together, these findings suggest that the PIC activation in the motoneuron dendrites may differentially affect the force production of the motor unit, depending not only on the firing state history of the motoneuron and the variation in muscle length but also on the mode of motor activity.NEW & NOTEWORTHY Cav1.3 channels in motoneuron dendrites are actively involved during normal motor activities. To investigate the effects of the activation of motoneuron Cav1.3 channels on force production, a model motor unit was built based on best-available data. The simulation results suggest that force potentiation induced by Cav1.3 channel activation is strongly modulated not only by firing history of the motoneuron but also by length variation of the muscle as well as neuromodulation inputs from the brainstem.

Motoneuron responsiveness to corticospinal tract stimulation during the silent period induced by transcranial magnetic stimulation.

Based on H-reflex data, spinal mechanisms are proposed to be responsible for the first 50-80 ms of the transcranial magnetic stimulation (TMS)-induced silent period. As several methodological issues can compromise H-reflex validity as a measure of motoneuron excitability, this study used transmastoid stimulation to elicit cervicomedullary motor evoked potentials (CMEPs) during the silent period. Eleven subjects made 1-3 visits which involved 32 or 44 brief (~3 s) isometric elbow flexor contractions at 25 % of maximal torque. During each contraction, transmastoid stimulation was delivered in isolation to elicit an unconditioned CMEP and at interstimulus intervals (ISIs) ranging from 50 to 150 ms after TMS to elicit a conditioned CMEP. Stimulus intensities for TMS and transmastoid stimulation were set to elicit a silent period of ~200 ms and an unconditioned CMEP of 15, 50, or 85 % of the maximal compound muscle action potential (M max), respectively. At all ISIs and intensities of transmastoid stimulation, the conditioned CMEP was significantly smaller than the unconditioned CMEP (p < 0.001). However, suppression of the conditioned CMEP was significantly less at 85 % compared to 15 or 50 % M max (p = 0.001). Contrary to published H-reflex data, the conditioned CMEP did not recover within 50-80 ms, remaining significantly suppressed at the longest ISI tested (150 ms). These data suggest the spinal portion of the TMS-evoked silent period is considerably longer than reported previously. Transmastoid stimulation, unlike peripheral nerve stimulation, does not impact proprioceptive inflow to motoneurons. Hence, relative to the H-reflex, the CMEP will be subjected to greater afferent-mediated disfacilitation and inhibition due to the TMS-induced muscle twitch.

Mild Hyperbaric Oxygen Improves Decreased Oxidative Capacity of Spinal Motoneurons Innervating the Soleus Muscle of Rats with Type 2 Diabetes.

Rats with type 2 diabetes exhibit decreased oxidative capacity, such as reduced oxidative enzyme activity, low-intensity staining for oxidative enzymes in fibers, and no high-oxidative type IIA fibers, in the skeletal muscle, especially in the soleus muscle. In contrast, there are no data available concerning the oxidative capacity of spinal motoneurons innervating skeletal muscle of rats with type 2 diabetes. This study examined the oxidative capacity of motoneurons innervating the soleus muscle of non-obese rats with type 2 diabetes. In addition, this study examined the effects of mild hyperbaric oxygen at 1.25 atmospheres absolute with 36 % oxygen for 10 weeks on the oxidative capacity of motoneurons innervating the soleus muscle because mild hyperbaric oxygen improves the decreased oxidative capacity of the soleus muscle in non-obese rats with type 2 diabetes. Spinal motoneurons innervating the soleus muscle were identified using nuclear yellow, a retrograde fluorescent neuronal tracer. Thereafter, the cell body sizes and succinate dehydrogenase activity of identified motoneurons were analyzed. Decreased succinate dehydrogenase activity of small-sized alpha motoneurons innervating the soleus muscle was observed in rats with type 2 diabetes. The decreased succinate dehydrogenase activity of these motoneurons was improved by mild hyperbaric oxygen. Therefore, we concluded that rats with type 2 diabetes have decreased oxidative capacity in motoneurons innervating the soleus muscle and this decreased oxidative capacity is improved by mild hyperbaric oxygen.

Arm-cycling sprints induce neuromuscular fatigue of the elbow flexors and alter corticospinal excitability of the biceps brachii.

We examined the effects of arm-cycling sprints on maximal voluntary elbow flexion and corticospinal excitability of the biceps brachii. Recreationally trained athletes performed ten 10-s arm-cycling sprints interspersed with 150 s of rest in 2 separate experiments. In experiment A (n = 12), maximal voluntary contraction (MVC) force of the elbow flexors was measured at pre-sprint 1, post-sprint 5, and post-sprint 10. Participants received electrical motor point stimulation during and following the elbow flexor MVCs to estimate voluntary activation (VA). In experiment B (n = 7 participants from experiment A), supraspinal and spinal excitability of the biceps brachii were measured via transcranial magnetic and transmastoid electrical stimulation that produced motor evoked potentials (MEPs) and cervicomedullary motor evoked potentials (CMEPs), respectively, during a 5% isometric MVC at pre-sprint 1, post-sprint 1, post-sprint 5, and post-sprint 10. In experiment A, mean power output, MVC force, potentiated twitch force, and VA decreased 13.1% (p < 0.001), 8.7% (p = 0.036), 27.6% (p = 0.003), and 5.6% (p = 0.037), respectively, from pre-sprint 1 to post-sprint 10. In experiment B, (i) MEPs decreased 42.1% (p = 0.002) from pre-sprint 1 to post-sprint 5 and increased 40.1% (p = 0.038) from post-sprint 5 to post-sprint 10 and (ii) CMEPs increased 28.5% (p = 0.045) from post-sprint 1 to post-sprint 10. Overall, arm-cycling sprints caused neuromuscular fatigue of the elbow flexors, which corresponded with decreased supraspinal and increased spinal excitability of the biceps brachii. The different post-sprint effects on supraspinal and spinal excitability may illustrate an inhibitory effect on supraspinal drive that reduces motor output and, therefore, decreases arm-cycling sprint performance.

Axon-somatic back-propagation in detailed models of spinal alpha motoneurons.

Antidromic action potentials following distal stimulation of motor axons occasionally fail to invade the soma of alpha motoneurons in spinal cord, due to their passing through regions of high non-uniformity. Morphologically detailed conductance-based models of cat spinal alpha motoneurons have been developed, with the aim to reproduce and clarify some aspects of the electrophysiological behavior of the antidromic axon-somatic spike propagation. Fourteen 3D morphologically detailed somata and dendrites of cat spinal alpha motoneurons have been imported from an open-access web-based database of neuronal morphologies, NeuroMorpho.org, and instantiated in neurocomputational models. An axon hillock, an axonal initial segment and a myelinated axon are added to each model. By sweeping the diameter of the axonal initial segment (AIS) and the axon hillock, as well as the maximal conductances of sodium channels at the AIS and at the soma, the developed models are able to show the relationships between different geometric and electrophysiological configurations and the voltage attenuation of the antidromically traveling wave. In particular, a greater than usually admitted sodium conductance at AIS is necessary and sufficient to overcome the dramatic voltage attenuation occurring during antidromic spike propagation both at the myelinated axon-AIS and at the AIS-soma transitions.

Inhibition of the primary motor cortex can alter one's "sense of effort": effects of low-frequency rTMS.

Studies using force-matching tasks have suggested that when we feel a "sense of effort," cortical regions may act to increase motor commands, and thus recruit additional motor units, in order to compensate for the exerted force. We hypothesized that suppressing activity in the primary motor cortex (M1), which is the source of the motor commands, would initiate the same process, and induce the same sense of effort. In a force-matching task, grip force was applied to 'right' hand and 10 healthy participants were asked to try to exert the same amount by using 'left' hand, with no visual feedback. On some trials, low-frequency, repetitive transcranial magnetic stimulation (lf-rTMS) was used to suppress the M1 and the primary somatosensory cortex (SI) in the left hemisphere, separately. Results showed that participants tended to overestimate the level of exerted force by up to 24%. In contrast, sham stimulation of the M1 and lf-rTMS over the SI did not significantly affect participants' estimations. Further, the M1 suppression resulted in a 42% reduction in motor-evoked potentials. Thus, the M1 suppression can affect our sense of effort, suggesting that compensatory neural mechanisms that increase the MI activity may play an important role in producing senses of effort.

Comparative Study of the L5 Spinal Nerve Transection Model and Sciatic Nerve Axotomy Model as a Peripheral Nerve Injury Model in Rat / 체질인류학회지

The aim of this study was to propose new more reliable peripheral nerve transection model to overcome the defect of the traditional sciatic axotomy model by specifically transecting L5 spinal nerve just after emerging from the intervertebral foramen and confining analysis area to the L5 spinal segment. The adult male Sprague-Dawley rats, weighing 300~350 g at the time of surgery, were used for the experiments. Four different experimental groups were used. 1. Sciatic nerve transection (Sc-Tx) group: transect the sciatic nerve in the popliteal fossa where it divided into the common peroneal nerve and tibial nerve. 2. L5 spinal nerve transection (L5-Tx) group: L5 spinal nerve was specifically transected. 3. Suture (Su) group: L5 spinal nerve was transected and immediately sutured. 4. Control group: the same surgical procedure with L5 spinal nerve transection group was performed except for the excision of L5 spinal nerve. To distinguish L5 motoneurons from the other level ones, the animals were received the retrograde tracer, FluoroGold into the axotomized proximal nerve stump. Serial coronal frozen sections at 40 microm thick through the L4 to L6 spinal segment was performed and the resultant total number of sections was about 180. Approximate serial 50 sections (approximately 2 mm) could be considered as the L5 segment based on the number of the fluorescent signals (above 20). L5 spinal segment could be differentiated from L4 and L6 segment based on their morphological characteristics under Cresyl violet stain. In L5-Tx group, at 2 and 4 weeks post-transection, the number of L5 spinal motoneurons was reduced by 8%. Meanwhile, Sc-Tx and Su groups showed no statistically notable changes. In this study, the authors could propose more reliable peripheral nerve axotomy model than the conventional sciatic nerve axotomy model by specifically transecting L5 spinal nerve and confining the investigating area within the L5 spinal segment.

Comparative Study of the L5 Spinal Nerve Transection Model and Sciatic Nerve Axotomy Model as a Peripheral Nerve Injury Model in Rat / 체질인류학회지

The aim of this study was to propose new more reliable peripheral nerve transection model to overcome the defect of the traditional sciatic axotomy model by specifically transecting L5 spinal nerve just after emerging from the intervertebral foramen and confining analysis area to the L5 spinal segment. The adult male Sprague-Dawley rats, weighing 300~350 g at the time of surgery, were used for the experiments. Four different experimental groups were used. 1. Sciatic nerve transection (Sc-Tx) group: transect the sciatic nerve in the popliteal fossa where it divided into the common peroneal nerve and tibial nerve. 2. L5 spinal nerve transection (L5-Tx) group: L5 spinal nerve was specifically transected. 3. Suture (Su) group: L5 spinal nerve was transected and immediately sutured. 4. Control group: the same surgical procedure with L5 spinal nerve transection group was performed except for the excision of L5 spinal nerve. To distinguish L5 motoneurons from the other level ones, the animals were received the retrograde tracer, FluoroGold into the axotomized proximal nerve stump. Serial coronal frozen sections at 40 microm thick through the L4 to L6 spinal segment was performed and the resultant total number of sections was about 180. Approximate serial 50 sections (approximately 2 mm) could be considered as the L5 segment based on the number of the fluorescent signals (above 20). L5 spinal segment could be differentiated from L4 and L6 segment based on their morphological characteristics under Cresyl violet stain. In L5-Tx group, at 2 and 4 weeks post-transection, the number of L5 spinal motoneurons was reduced by 8%. Meanwhile, Sc-Tx and Su groups showed no statistically notable changes. In this study, the authors could propose more reliable peripheral nerve axotomy model than the conventional sciatic nerve axotomy model by specifically transecting L5 spinal nerve and confining the investigating area within the L5 spinal segment.

Effects of hypoxic exposure and endurance exercise training on the oxidative potentials of soleus muscle fibers and motoneurons in the rat / 体力科学

Thirty (n=30) seven week old male Sprague-Dawley rats were divided into six groups of five rats (n=5) in each group. The groups were designated Sc=sea level controls; St=sea level trained; Fc=hypoxic exposed (16% O<SUB>2</SUB>) controls; Ft =hypoxic exposed (16% O<SUB>2</SUB>) trained; Pc=intermittent hypoxic exposed (18%, 16%, 14%, 16%, 18% O<SUB>2</SUB> for two days each) controls; and Pt=intermittent hypoxic exercise trained. Exercise training consisted of 45min/day running on a rat treadwheel for 24 consecutive days. Fiber type distribution, succinate dehydrogenase (SDH) activity and glycogen content of the soleus muscle and the oxidative enzyme activity of the motoneurons of the soleus were measured in each group after the 24 days of hypoxic exposure and exercise training. In comparison to each training group's control the glycogen concentration of the soleus muscle was increased (P<0.05) regardless of hypoxic exposure. Only the intermittently hypoxic exercise trained group (Pt) demonstrated a fiber type shift of slow-twitch oxidative to fast-twitch oxidative glycolytic fibers. Neither hypoxia or exercise training altered the oxidative enzyme capacity of the soleus motoneurons.