Naomi Wada

Morphological changes in the masseter muscle and its motoneurons during postnatal development.

It has been suggested that the morphological properties of the masseter muscle are changed by the masticatory activity pattern. In the rat, the activity pattern of the muscle alters from sucking to biting around 3 weeks after birth. The working hypothesis in this study is that the unique alteration in masticatory activity has an important influence on the development of the masseter muscle and its motoneurons.

Characteristics of H- and M-waves recorded from rat forelimbs

The Hoffman reflex (H-reflex) is a useful tool for studying the functional aspects of the spinal cord without anesthesia and/or damage to the body. H-reflex studies are performed mainly in the hindlimbs. The purpose of the present study was to evaluate the characteristics of the H-reflex in the forelimbs and hindlimbs in rats anesthetized with ketamine-HCl. H- and M-waves were recorded from the interosseous muscles after electrical stimulation of the n. lateral plantar of the hindlimb and n. medialis of the forelimb. Hmax/Mmax values were significantly smaller in the forelimbs than in the hindlimbs. Furthermore, paired-pulse attenuation tended to be stronger in the forelimbs than in the hindlimbs. These findings suggest that control by descending and/or propriospinal pathways is stronger in the forelimbs than in the hindlimbs in rats.

The role of vertebral column muscles in level versus upslope treadmill walking—An electromyographic and kinematic study

To gain insight into the neural mechanisms controlling vertebral column movement and its role in walking, we performed kinematic and electromyographic (EMG) studies on cats during level and upslope treadmill walking. Kinematic data of the limbs and vertebral column were obtained with a high-speed camera synchronized with EMG recordings from levels T10, L1, and L5 of m. longissimus dorsi (Long). During a single-step cycle at all upslope angles, vertebral movement in the lateral (left-right), cranial-caudal (forward-backward), and dorsal-ventral (upward-downward) directions was observed. Lateral movements were produced by forelimb take-off and hindlimb landing, and forward and upward movements were produced by hindlimb extension. During the single-step cycle, each of the three epaxial muscles, m. multifidus, m. iliocostalis, and Long, showed two bilateral EMG bursts. The onset of the EMG bursts coincided with the left-right movements, suggesting that epaxial muscle activity depresses lateral movement. The termination of the EMG bursts correlated with the forward and downward phase of the step cycle, suggesting that contraction of the epaxial muscles produces forward and downward movements. EMG bursts of the epaxial muscles increase the stiffness and produce inwardly movements to decrease the lateral movements of the vertebral column and the termination of EMG bursts control the movements into cranial and ventral direction of the vertebral column. The results suggest that the rhythmic EMG bursts in the epaxial muscles are produced by pattern generators, and the timing of EMG bursts among the different levels of the epaxial muscles are altered by walking condition input via peripheral afferents and descending pathways.

Neuronal pathways for spinal reflexes activated by group I and group II muscle afferents in the spinal segment (Co1) innervating the tail in the low spinalized cat

Abstract We studied neuronal pathways for spinal reflexes activated by group-I and group-II muscle afferents in the spinal segments innervating the tail in unanesthetized and spinalized (L1) cats. Experiments were performed on 25 adult cats of both sexes. The effects of stimulating nerves innervating six tail muscles on both sides were recorded from tail motoneurons in the first coccygeal spinal segment (Co1) using glass microelectrodes. Stable recordings were obtained from 150 tail motoneurons. Stimulation of group-I muscle afferents (stimulus intensity <1.8 T) often produced EPSPs (82/150) after stimulating nerves innervating neighboring tail muscles. Motoneurons innervating the long-tendoned muscles, M. extensor caudae lateralis and M. flexor caudae longus (ECL and FCL), received heteronymous monosynaptic connections from group-I muscle afferents innervating the ipsilateral tail muscles. The motoneurons innervating segmental muscles, M. extensor caudae medialis and M. flexor caudae brevis (ECM and FCB), received heteronymous monosynaptic connections from group-I muscle afferents innervating tail muscles on both sides. The motoneurons innervating tail muscles originated from the Ossa coxae, M. abductor caudae externus and M. abductor caudae internus (ACE and ACI), received monosynaptic connection from group-I muscle afferents innervating most of the tail muscles on both sides. Crossed disynaptic inhibitory pathways activated by primary muscle afferent inputs were observed in ECM, ACE, FCL, and FCB motoneurons. The effects of group-II afferent inputs were not dependent on the kind of motoneuron, and alternative excitatory and inhibitory pathways were not clearly observed in the tail motoneuron pool. It is suggested that variability of the neuronal pathways from group-I and -II muscle afferents to tail motoneurons corresponds to functional relationships among tail muscles, depending on the tail movements.

Neuronal pathways from low-threshold hindlimb cutaneous afferents to motoneurons innervating trunk muscles in low-spinal cats

Abstract Postsynaptic potentials (PSPs) evoked in motoneurons innervating the back and abdominal muscles in the lumbar part of the body by stimulating hindlimb cutaneous afferents were investigated in unanesthetized decerebate and spinal cats. Various types of PSP: pure excitatory postsynaptic potential (EPSP), pure inhibitory postsynaptic potential (IPSP), and mixed PSP (i.e., EPSP followed by IPSP, EPSP/IPSP; and IPSP followed by EPSP, IPSP/EPSP) were observed. The weak stimulation at 2 times threshold (2T) produced predominantly the EPSP, while at 5T the incidence of IPSP or EPSP followed by IPSP was increased. In about 20–50% of the various groups of motoneurons, PSPs evoked by ipsi- and contralateral nerves were qualitatively and quantitatively similar. For the other motoneurons, PSPs evoked by ipsi- and contralateral nerves were markedly different with respect to magnitude and/or polarity. These findings suggest that, within each motoneuron pool, some neurons act to increase stiffness of the trunk or to move vertically in response to an increased activity of cutaneous afferents, while the other motoneurons act to produce lateral bending of the trunk.

Expression mapping of cytotoxic T-lymphocyte antigen-2α gene transcripts in mouse brain

Cytotoxic T-lymphocyte antigen-2α (CTLA-2α), an inhibitor peptide homologous to the proregion of mouse cathepsin L, was originally discovered and expressed in mouse-activated T-cells and mast cells. Expressed recombinant CTLA-2α is shown to exhibit selective inhibition to cathepsin L-like cysteine proteinases. However, its in vivo targets in mammalian tissues are yet to be identified. We carried out in situ hybridization studies to examine the expression pattern of CTLA-2α mRNA and determine the specific cell types synthesizing CTLA-2α in the mouse brain. CTLA-2α mRNA was detected in various neuronal populations within the telencephalon in cerebral cortices, olfactory system, septum, basal ganglia, amygdala and highest levels were observed in the hippocampus. Within the diencephalon high density of positive cells was found in mediodorsal and lateral posterior thalamic nuclei and medial habenular nucleus (MHb). In the hypothalamus, high density of CTLA-2α mRNA labeling was seen in the suprachiasmatic nucleus (Sch), optic tract, arcuate nucleus, and median eminence. The fasciculus retroflexus and its termination in the mesencephalic interpeduncular nucleus were also densely labeled. Other mesencephalic expression sites were the superior colliculus, periaqueductal gray, paramedian raphe nucleus, and inferior colliculus. In the rhombencephalon, strong labeling was detected in the pontine, vestibular, and reticular nuclei. Intense expression was also noted within cerebellar cortex in Purkinje neurons and at a moderate level in granule cell layer, stellate, and basket cells. A possible function of this novel inhibitor peptide in relation to learning, memory, and diseases is discussed.

Dendritic and axonal localization of cytotoxic T-lymphocyte antigen-2 alpha protein in mouse brain

Cytotoxic T-lymphocyte antigen-2 alpha (CTLA-2alpha) is a novel cysteine proteinase inhibitor protein originally discovered and expressed in mouse activated T-cells and mast cells. Expressed recombinant CTLA-2alpha is shown to exhibit selective inhibition of cathepsin L-like cysteine proteinases. We have recently reported the expression pattern of CTLA-2alpha mRNA in mouse brain by in situ hybridization, demonstrating that it is mainly enriched within neuronal populations. In this study we present the distribution profile of the protein by immunohistochemical analysis. Results showed that CTLA-2alpha protein is preferentially localized in dendritic and axonal compartments. In telencephalon, strong labeling was detected in dendrites in the cerebral cortices, stratum radiatum and stratum lacunosum moleculare and within axonal fibers of stratum lucidum where mossy fibers emanating from all parts of the granule cell layer of dentate gyrus terminate at pyramidal neurons and interneurons. In diencephalon, moderate staining was found in all thalamic nuclei but was strong in medial habenular nucleus and the hypothalamic nuclei including suprachiasmatic nucleus, optic chiasm, arcuate nucleus and median eminence. In mesencephalon, strong immunoreactivity was detected in superior colliculus, inferior colliculus and paramedian raphe nucleus. In the rhombencephalon, the pontine nucleus and transverse fibers of the pons revealed strong staining but were moderate in vestibular nuclei. Strong immunoreactivity was also observed in the internal white matter, granule cell layer and Purkinje cell layer within cerebellum. On Western blot analysis, a band of 14 kDa for CTLA-2alpha from protein extracts of the cerebrum, cerebellum, pons and medulla was detected. The distribution pattern and functional considerations of CTLA-2alpha in the brain are discussed.

Neuronal pathways from low-threshold muscle and cutaneous afferents innervating tail to trunk muscle motoneurons in the cat.

Abstract. We studied neuronal pathways from low-threshold muscle (group I, II) and cutaneous afferents (group Aαβ) innervating the tail to motoneurons innervating trunk muscles (m. iliocostalis lumborum and m. obliquus externus abdominus) in 18 spinalized cats. Stimulation of group I muscle afferents produced excitatory postsynaptic potentials or excitatory postsynaptic potentials followed by inhibitory postsynaptic potentials in all motoneurons innervating the m. iliocostalis lumborum which showed effects (32%), and predominantly inhibitory postsynaptic potentials in motoneurons innervating the m. obliquus externus abdominus (47%). Stimulation of group I+II afferents produced significant increases of the incidence of motoneurons showing postsynaptic potentials (the motoneurons innervating the m. iliocostalis lumborum, 87%; the motoneurons innervating the m. obliquus externus abdominus, 82%). The effects of low threshold cutaneous afferents were bilateral, predominantly producing inhibitory postsynaptic potentials in motoneurons innervating both muscles. These results suggest that neuronal pathways from muscle afferents to back muscle motoneurons mainly increase the stiffness of the trunk to maintain its stability, while those to abdominal muscles help to extend the dorsal column by decreasing their activities. The results also indicate that neuronal pathways from cutaneous afferents to trunk motoneurons functionally disconnect the tail from the trunk.

Electromyographic activity of m. longissimus and the kinematics of the vertebral column during level and downslope treadmill walking in cats.

Electromyographic (EMG) burst patterns of m. longissimus and the kinematics of the vertebral column were assessed in cats during treadmill walking for six downslope grades (5 degrees-30 degrees). The EMG bursts during downslope walking were weak between 5 degrees and 20 degrees. At steeper grades (>20 degrees), EMG bursts were large. Bursts at T10 facilitated inward movements, and those at L1 decreased forward movements, while those at L5 decreased backward movements during downslope walking at steeper grades.

Hoffmann reflex in a rat bipedal walking model

The rat bipedal walking model (RBWM) refers to rats that acquired anatomical and functional characteristics for bipedal walking after the completion of a long-term motor training program. We recorded the Hoffmann reflex (H-reflex) of the forelimb and hindlimb in RBWM and control (not trained, normal) rats to evaluate the effects of bipedal walking on central nervous system (CNS) activity. The H-reflex recorded from the hindlimbs of the RBWM was significantly inhibited compared with that in the control. Furthermore, the inhibition of the H-reflex recorded from both forelimbs and hindlimbs by paired pulse stimulation tended to be enhanced in RBWM. These results indicate that bipedal walking or bipedal walking training cause functional changes in spinal reflex pathways in the CNS.