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M Mishina

Publications and source records attributed to M Mishina.

At least 127 records · Page 7Linked to original sources
PubMed1995

[Changes in regional cerebral blood flow in nondominant hemisphere during speech task in aphasics.: a PET activation study].

To investigate the patients with language disorder and to detect the activated areas in language processing, we measured the changes in the regional cerebral blood flow (CBF) during speech task (¿counting¿ task) using O-15 water PET activation technique in six normal subjects (age 58.3 +/- 8.1), mean +/- SD), ten fluent aphasics (age 60.3 +/- 12.5) and six nonfluent aphasics (age 50.5 +/- 8.3). In ¿counting¿ task, the subjects were instructed to count the number aloud from 1 to 10 and repeat the sequence over and over in the native language (Japanese) at the rate of one number per 2.5 sec. The data were analyzed with stereotactic intersubject averaging analysis for the normal subjects. Apart from it, the regions of interest (ROI) analysis (a circular ROI of 12 mm diameter) was performed in the language-related areas: posteroinferofrontal areas (PIF), posterosuperotemporal area (PST), and Rolandic areas (related to the mouth and lips) were significantly activated bilaterally in the ¿counting¿ task. PIF were activated with a dominance in the left side. In the resting state, rCBF in the left PIF and left PST were activated with a dominance in the left side. In the resting state, rCBF in the left PIF and left PST was reduced in both fluent and nonfluent aphasics. In nonfluent aphasics, the magnitude of activation in the right PIF by ¿counting¿ task was significantly greater than normal subjects and patients with fluent aphasia (ANOVA). It suggests the importance of the right PIF in the simple language processing in the nonfluent aphasia. In the sixteen aphasic patients, the increase in rCBF in the right PST during the ¿counting¿ task was negatively correlated with the score of comprehension (WAB) in the Spearman ranked correlation (p < 0.05). It suggests that, while the activity of the right PST is minimal for the ¿counting¿ in normal subjects, it plays an important and compensatory role in aphasic patients.

Adult↗
PubMed1995

[Regional cerebral glucose metabolism associated with ataxic gait--an FDG-PET activation study in patients with olivopontocerebellar atrophy].

We studied changes in regional cerebral glucose metabolism during bipedal gait in 7 patients with olivopontocerebellar atrophy (OPCA) and 7 control subjects without neurological disease. All OPCA patients in this study were able to walk on the treadmill without a support or handrails. We used 18F-2-fluoro-2-deoxy-D-glucose (18F-FDG) and positron emission tomography (PET) to evaluate glucose metabolism under two different conditions; 30 minutes' treadmill walking, and supine resting. The two sets of PET images were three-dimensionally registered to the magnetic resonance image (MRI) in each subject. Then, the PET images were normalized by the global value. Regions of interest (ROIs) were drawn on the cerebellar vermis, cerebellar hemispheres, pons, and thalamus, and FDG uptake was obtained to calculate the activation ratio (= [FDG uptake under walking]/[FDG uptake under resting]) for each region. We used unpaired t-test for statistical analysis to compare the OPCA patients and the normal control subjects. Normalized resting FDG uptake had no significant difference between control subjects and OPCA patients in any region. Activation ratio of OPCA patients was significantly decreased in the cerebellar vermis compared with the control subjects. In the control subjects, FDG uptake had little difference between resting and walking in the cerebellar hemisphere, pons and thalamus. On the other hand, the FDG uptake of OPCA patients was moderately increased by walking in these regions. In the cerebellar vermis, in spite of no significant difference between the normals and the patients during resting, the activation ratio in OPCA patients was significantly decreased. We speculate that the reduction of activation ratio in the cerebellar vermis reflects the dysfunction caused by degeneration. The result suggests that the PET activation study can demonstrate cerebellar dysfunction in the early phase of OPCA, in which other neuro-imaging methods cannot detect the tissue atrophy, hypometabolism or hypoperfusion in the resting state. In the cerebellar hemisphere, pons and thalamus, the activation ratio was nearly equal to one in control subjects, while it was larger in OPCA patients. We speculate that the instability during the ataxic gait increases the inputs from the vestibular, somatosensory and visual systems to these regions and outputs from these regions to the other neural systems. In conclusion, PET activation study is a useful and noninvasive technique for investigating the brain function associated with human gait.

Adult↗
PubMed1994

Expression and characterization of the alpha 2 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-selective glutamate receptor channel in a baculovirus system.

Using a baculovirus expression vector system, the alpha 2 subunit of the mouse alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA)-selective glutamate receptor (GluR) channel was expressed in Spodoptera frugiperda insect cells. Immunoblotting using the antibody made to the synthetic peptide corresponding to the C-terminus of GluR alpha 2 and [35S]methionine/[35S]cysteine metabolic radiolabeling revealed the major 102-kDa and the minor 98-kDa protein bands. Metabolic radiolabeling with tunicamycin suggested that the two bands correspond to glycosylated and unglycosylated forms, respectively. The recombinant GluR alpha 2 proteins expressed in insect cells were also identified by immunofluorescence staining. The results of [3H]AMPA binding assay using whole cells suggested that, in infected Sf21 cells, binding sites of the GluR alpha 2 proteins were possibly located on the extracellular side. Scatchard analysis of AMPA binding showed the following parameters: Kd = 16 nM, Bmax = 1.9 x 10(5) binding sites per cell or 1 pmol/mg protein in the total particulate fraction. The ligand binding characteristics of the receptors expressed in insect cells were examined. From the effect of various agonists on [3H]AMPA binding of the receptors expressed in insect cells, the rank order potency of agonists was quisqualate > AMPA > L-glutamate > kainate. Thus, the baculovirus-insect cell expression system provides high-efficiency expression of the receptor sufficient to permit structural and functional analyses.

Animals↗
PubMed1994

Involvement of the glutamate receptor delta 2 subunit in the long-term depression of glutamate responsiveness in cultured rat Purkinje cells.

An antisense oligonucleotide against the glutamate receptor delta 2 subunit mRNA, which is selectively expressed only in Purkinje neurons, suppressed the induction of long-term depression (LTD) of glutamate responsiveness in the rat cerebellar culture. LTD of glutamate response is induced by pairing glutamate application and depolarization of a Purkinje cell. Treatment of the culture with the antisense oligonucleotide exerted no appreciable effect on basic physiological and morphological properties of Purkinje cells, except for LTD induction and reduction of delta immunoreactivity which was intense in distal dendrites. Sense and missense oligonucleotides, which were used as controls, did not block LTD induction. These results suggest that the glutamate receptor delta 2 subunit is involved in the cerebellar LTD.

Action Potentials↗
PubMed1994

Ligand-binding properties and N-glycosylation of alpha 1 subunit of the alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate(AMPA)-selective glutamate receptor channel expressed in a baculovirus system.

The alpha 1 subunit of the mouse alpha-amino-3-hydroxy-5-methyl-4-isoxazole- propionate(AMPA)-selective glutamate receptor channel has been expressed in insect Spodoptera frugiperda cells using a baculovirus system. The recombinant receptor proteins were identified by immunocytochemical detection, Western-blot analysis, and [35S]methionine/[35S]cysteine metabolic labeling experiments. The effect of tunicamycin on the metabolic labeling and immunoblots suggested that the two products, a major protein species of approximately 104 kDa and a minor species of approximately 100 kDa, correspond to glycosylated and non-N-glycosylated forms, respectively, which was also supported by the enzymic deglycosylation experiments. The lack of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate-binding activity of non-N-glycosylated glutamate receptor expressed in the presence of tunicamycin suggested that N-glycosylation is required, directly or indirectly, for functional expression in insect cells for ligand binding. Scatchard analysis of alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate binding showed a single binding site with Kd 30 nM and a Bmax value of 2.6 x 10(5) binding sites/cell or 1.5 pmol/mg protein in the total particulate fraction. Among the compounds tested in the competition studies, beta-(3,5-dioxo-1,2,4-oxadiazolidin-2-yl)-L-alanine (quisqualate) was the most potent inhibitor of the 3H-labeled alpha-amino-3-hydroxy-5-methyl-4-isoxazole-propionate binding (IC50 = 30 nM), followed in decreasing order by alpha-amino-3-hydroxy-5- methyl-4-isoxazole propionate, L-glutamate, 6,7-dinitroquinoxaline-2,3-dione, 6-cyano-7-nitroquinoxaline-2,3-dione, and 2-carboxy-4-(1-methylethenyl)-3-pyrrolidineacetate (kainate). Thus, in this study we present detailed analysis of alpha-amino-3-hydroxy-5-methyl-4- isoxazole-propionate-binding activity of the homomeric (single subunit) glutamate receptor channel of mouse alpha 1 subunit and discuss possible roles of N-glycosylation of the glutamate receptor channel alpha 1 subunit.

Amino Acid Sequence↗
PubMed1994

Distinct spatiotemporal distributions of the N-methyl-D-aspartate receptor channel subunit mRNAs in the mouse cervical cord.

The distribution of five N-methyl-D-aspartate (NMDA) receptor channel subunit mRNAs in the mouse spinal cord from embryonic day 13 (E13) through postnatal day 56 (P56) was semiquantitatively examined at the cervical level via in situ hybridization with subunit-specific oligonucleotide probes. Signals for the epsilon 1 subunit mRNA were restricted to the most ventral portion of the spinal cord during embryonic stages. They extended to all laminae of the spinal cord except for the lamina 2 (substantia gelatinosa) during postnatal development. A wide expression of the epsilon 2 subunit mRNA was found in the spinal gray matter from E13 through neonatal stages, but the signals became restricted to the lamina 2 by P21. No significant signals for the epsilon 3 subunit mRNA were detected in the spinal cord at any developmental stages. The epsilon 4 subunit mRNA was distributed widely in the spinal cord during embryonic and early postnatal periods but decreased nearly to background levels by P21. In contrast to the differential distribution of the epsilon subunit mRNAs, the zeta 1 subunit mRNA was found ubiquitously at each developmental stage examined. These findings suggest that the molecular organization of the epsilon subunits may be different between the dorsal horn and the remaining regions in the mature spinal cord, which provides a molecular basis for functional heterogeneity of the NMDA receptor channel. Moreover, this spatial heterogeneity might be generated through drastic alterations in the subunit composition of the channel complex during spinal cord development.

Animals↗
PubMed1994

Distinct spatiotemporal expressions of five NMDA receptor channel subunit mRNAs in the cerebellum.

The distribution of five NMDA receptor channel subunit mRNAs was examined in the mouse cerebellum from embryonic day 13 through postnatal day 56, by in situ hybridization with subunit-specific oligonucleotide probes. At postnatal days 21 and 56, each cerebellar neuron displayed differential expressions of the epsilon subunit mRNAs. The granule cells showed hybridizing signals for the epsilon 1 and epsilon 3 subunit mRNAs, the molecular layer neurons for the epsilon 4 subunit mRNA, and the cerebellar nucleus neurons for the epsilon 1 and epsilon 4 subunit mRNAs, whereas the Purkinje cells did not express any epsilon subunit mRNAs. At early postmitotic stages of development, the epsilon 2 subunit mRNA appeared in each cerebellar neuron, including the Purkinje cells, and the epsilon 4 subunit mRNA appeared in neurons of the molecular layer and the cerebellar nuclei. The expression patterns in the cerebellum altered drastically during the first 2 postnatal weeks; the epsilon 1 and epsilon 3 subunit mRNAs appeared in the granule cells and the cerebellar nucleus neurons, whereas the epsilon 2 subunit mRNA disappeared from each neuron and the signal levels of the epsilon 4 subunit mRNA decreased remarkably. In contrast to the differential expressions of the four epsilon subunit mRNAs, intense signals for the zeta 1 subunit mRNA were observed in each cerebellar neuron from early postmitotic stages through the mature stage. These findings suggest that anatomical organization of the epsilon subunits is heterogeneous in the cerebellum both spatially and temporally, which would give rise to functional diversity of the NMDA receptor channel.

Aging↗
PubMed1994

Distinct distributions of five NMDA receptor channel subunit mRNAs in the brainstem.

The distribution of five NMDA receptor channel subunit mRNAs in the mouse brainstem at postnatal day 21 was semiquantitatively examined by in situ hybridization with subunit-specific oligonucleotide probes. The epsilon 1 subunit mRNA was observed in various brainstem nuclei. On the other hand, the epsilon 2 and epsilon 3 subunit mRNAs were restricted to particular nuclei, and the epsilon 4 subunit mRNA was detected in several nuclei at very low levels. The dorsal cochlear nucleus exhibited differential expression of the epsilon subunit mRNAs in distinct neuron types: the epsilon 2 subunit mRNA in the pyramidal cells, the epsilon 3 subunit mRNA in the granule cells, and the epsilon 1 subunit mRNA in other neurons. In the vestibular nuclei, the medial nucleus expressed the epsilon 1, epsilon 3, and epsilon 4 subunit mRNAs, whereas the lateral and superior nuclei expressed the epsilon 1 subunit mRNA. Such region-specific expressions were also discerned in the central gray, the superior and inferior colliculi, the medial accessory oculomotor nucleus, the locus ceruleus, the parabrachial nucleus, nucleus of the solitary tract, the caudal subnucleus of the trigeminal spinal tract nucleus, and the inferior olive. In contrast to the differential distributions of the four epsilon subunit mRNAs, intense signals for the zeta 1 subunit mRNA were distributed throughout the brainstem. These findings suggest that anatomical organization of the epsilon subunits is heterogeneous in various regions of the brainstem, which would give rise to functional diversity of the NMDA receptor channel in these regions.

Animals↗
PubMed1994

A novel mutation in Cu/Zn superoxide dismutase gene in Japanese familial amyotrophic lateral sclerosis.

Recently, several missense mutations in the Cu/Zn superoxide dismutase gene (SOD1) have been reported as a putative cause of chromosome-21q-linked familial amyotrophic lateral sclerosis (FALS). We have discovered a novel missense mutation (substitution of Thr for Ala4) in exon 1 (GCC to ACC) in two FALS patients from one Japanese FALS family. No mutations were found in 17 cases of sporadic ALS. The enzyme activity of recombinant fusion protein containing the Cu/Zn superoxide dismutase (SOD) with the Ala4-to-Thr mutation was significantly reduced in E. coli. On the other hand, in the expression system in insect cells using Baculovirus, the mutant SOD expressed an enzyme activity as high as wild-type SOD. These results suggest that the stability of SOD with the Ala4-to-Thr mutation is disrupted especially in the fusion protein. Autopsy was carried out on one of the two patients, and the pathological findings were typical of FALS with posterior column involvement. These results raise the possibility that mutation of the SOD1 is responsible for FALS with broader pathological involvement.

Amino Acid Sequence↗
PubMed1994

Circadian expression of NMDA receptor mRNAs, epsilon 3 and zeta 1, in the suprachiasmatic nucleus of rat brain.

The presence of the N-methyl-D-aspartate (NMDA) receptor channel subunit epsilon 3 and zeta 1 mRNAs in the rat suprachiasmatic nucleus (SCN) was detected by sensitive in situ hybridization. The daily fluctuations in the epsilon 3 and zeta 1 subunit mRNAs in their abundance were found in the SCN to be high during the day and lower during the night under 12 h light:12 h dark conditions (LD 12:12). Under constant darkness for 15 days, both the epsilon 3 and the zeta 1 mRNA levels in the SCN remained cyclic. Furthermore, after exposure of rats to light, the epsilon 3 and zeta 1 subunit mRNAs increased during the subjective night, but not during the subjective day. These results implicate the involvement of the epsilon 3 and zeta 1 subunits in neuronal signaling in the SCN and suggest that these subunits of the NMDA receptor channel are regulated by light and a circadian clock.

Animals↗
PubMed1994

Differential distributions of the NMDA receptor channel subunit mRNAs in the mouse retina.

In the retina, the epsilon 2 and zeta 1 subunit mRNAs of the NMDA receptor channel were expressed from embryonic stages and found in ganglion cell layer and whole layer of inner nuclear layer at postnatal day 21 (P21). The epsilon 1 subunit mRNA appeared postnatally and was distributed in ganglion cell layer and an inner third of inner nuclear layer at P21. These findings suggest that molecular organization of the NMDA receptor channel may alter during the retinal development.

Animals↗
PubMed1994

Distinct gene expression of the N-methyl-D-aspartate receptor channel subunit in peripheral neurons of the mouse sensory ganglia and adrenal gland.

The distribution of five N-methyl-D-aspartate (NMDA) receptor channel subunit mRNAs was examined in the mouse peripheral nervous tissues by in situ hybridization. The trigeminal and dorsal root ganglion cells showed signals for the zeta 1 subunit mRNA from embryonic stages through postnatal day 21, but not for the epsilon 1, epsilon 2, epsilon 3, and epsilon 4 subunit mRNAs at any developmental stages examined. The adrenal medulla also expressed the zeta 1 subunit mRNA alone. These findings suggest that molecular organization of the NMDA receptor channel in peripheral neurons are distinct from those in central neurons, which may result in different functional properties of the channel between them.

Adrenal Glands↗
PubMed1994

Familial amyotrophic lateral sclerosis with a mutation in the Cu/Zn superoxide dismutase gene.

Several missense mutations within exons 1, 2, 4 and 5 of the gene for Cu/Zn-binding superoxide dismutase (SOD1) have been discovered to be involved in the development of chromosome 21q-linked familial amyotrophic lateral sclerosis (FALS). We describe here an autopsied patient with FALS, in whom we have recently identified a novel missense mutation in exon 1 of the SOD1 gene. The neuropathological findings were compatible with those described previously in patients with FALS with posterior column involvement. This suggests that mutations of the SOD1 gene may be responsible for this form of FALS.

Adult↗
PubMed1994

Different spatio-temporal expressions of three otx homeoprotein transcripts during zebrafish embryogenesis.

Three zebrafish otx homeoproteins containing a homeodomain homologous to that of the Drosophila orthodenticle head gap gene product have been identified by cloning and sequencing of cDNAs. The zebrafish otx2 homeoprotein shares high amino-acid sequence identity with the mouse Otx2 homeoprotein, whereas the zebrafish otx1 and otx3 homeoproteins exhibit moderate homology with the mouse Otx1 and Otx2 homeoproteins. Three otx homeoprotein mRNAs show different spatio-temporal expression patterns during zebrafish embryogenesis as revealed by Northern blot and whole mount in situ hybridization analyses. Large amounts of the otx1 homeoprotein mRNA are found in fertilized uncleaving eggs. The otx3 homeoprotein mRNA appears in the embryonic shield, the site of the organizer. In the developing brain, three zebrafish otx mRNAs are distributed in the diencephalon and the midbrain, but their fine expression patterns are different. These results suggest that three zebrafish otx homeoproteins, alone or in combination, may play roles in very early embryogenesis, gastrulation, and the development and subdivision of the diencephalon and the midbrain.

Amino Acid Sequence↗
PubMed1994

Ion permeation properties of the cloned mouse epsilon 2/zeta 1 NMDA receptor channel.

The heteromeric mouse epsilon 2/zeta 1 N-methyl-D-aspartate (NMDA) receptor was expressed in Xenopus oocytes, and its channel properties were studied using both the outside-out patch-clamp and two-microelectrode voltage-clamp techniques. In the cloned receptor channel, permeation properties of monovalent and divalent cations, and voltage-dependent block by Mg2+ were similar to those reported previously in the native NMDA receptor channels. The sequence of single-channel conductances for alkali metals was Rb+ > Cs+ approximately K+ > Na+ > Li+, whereas the sequence of relative permeabilities was Cs+ > Rb+ > K+ approximately Na+ > Li+. The single-channel conductances measured in isotonic Ca2+, Sr2+ and Ba2+ solutions were almost equal, and approximately one-fifth of the value in the Na+ solution, although the permeabilities for these alkaline earth cations were higher than for Na+. It is likely that Ca2+, Sr2+ and Ba2+ would enter the NMDA receptor channel more easily than Na+, but would bind to a site in the channel more tightly, the net effect being a reduced value of the current.

Animals↗
PubMed1994

Circadian variation of urinary enzymes in the dog.

Circadian variations of N-acetyl-beta-D-glucosamidase (NAG, EC 3.2.1.30) and gamma-glutamyl transpeptidase (gamma-GTP, EC 2.3.2.2) were examined in dogs. Urine and blood were collected at 4-hr intervals from adult mongrel dogs (11 males, 11 females) weighing between 7 and 13 kg. The collected urine and blood were used to measure creatinine, NAG and gamma-GTP. The amount of urine was significantly higher (p < 0.05) during the period from 12:00 to 16:00. Creatinine clearance did not show any characteristic variations and creatinine excretion was significantly higher (p < 0.05) during the period from 8:00 to 16:00. No characteristic variations were observed in the amounts excreted, activity, or NAG and gamma-GTP creatinine index. There were high correlations between creatinine excretion and creatinine clearance (r = 0.693), gamma-GTP excretion and gamma-GTP creatinine index (r = 0.724), NAG excretion and NAG creatinine index (r = 0.878), and between the NAG and gamma-GTP creatinine index of each 4-hr specimen and the NAG and gamma-GTP excretion of the 24-hr specimens. These results suggest the diagnostic usefulness measuring enzyme activity in non-periodically collected urine.

Acetylglucosaminidase↗