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M Kano

Publications and source records attributed to M Kano.

At least 55 records · Page 3Linked to original sources
PubMed2000

L-serine and glycine serve as major astroglia-derived trophic factors for cerebellar Purkinje neurons.

Glial cells support the survival and development of central neurons through the supply of trophic factors. Here we demonstrate that l-serine (l-Ser) and glycine (Gly) also are glia-derived trophic factors. These amino acids are released by astroglial cells and promote the survival, dendritogenesis, and electrophysiological development of cultured cerebellar Purkinje neurons. Although l-Ser and Gly are generally classified as nonessential amino acids, 3-phosphoglycerate dehydrogenase (3PGDH), a key enzyme for their biosynthesis, is not expressed in Purkinje neurons. By contrast, the Bergman glia, a native astroglia in the cerebellar cortex, highly expresses 3PGDH. These data suggest that l-Ser and Gly mediate the trophic actions of glial cells on Purkinje neurons.

Animals↗
PubMed2000

Induction of protective immunity against pathogenic simian immunodeficiency virus by a foreign receptor-dependent replication of an engineered avirulent virus.

In AIDS vaccine strategies, live attenuated vaccines can confer good resistance against pathogenic virus infections but have the potential risk of inducing disease, whereas safer replication-negative strategies such as DNA vaccinations have so far failed to prevent the disease onset. Here, we developed a novel DNA vaccine strategy to induce restricted replication of an avirulent virus and evaluated it in a simian immunodeficiency virus (SIV) infection model. We generated a chimeric SIV, FMSIV, by replacing SIV env with ecotropic Friend murine leukemia virus (FMLV) env to confine its replication to FMLV receptor (mCAT1)-expressing cells. In primate cells lacking mCAT1, FMSIV did not replicate unless mCAT1 was introduced exogenously. Vaccination to macaques with both the FMSIV DNA and the mCAT1-expression plasmid DNA induced SIV Gag-specific cellular immune responses and resistance against pathogenic SIV(mac239) challenge more efficiently than the replication-negative control vaccination with the FMSIV DNA alone. This strategy may be useful for development of safe and effective vaccines against various kinds of pathogenic viruses.

AIDS Vaccines↗
PubMed2000

Calcium-dependent persistent facilitation of spike backpropagation in the CA1 pyramidal neurons.

Sodium-dependent action potentials initiated near the soma are known to backpropagate over the dendrites of CA1 pyramidal neurons in an activity-dependent manner. Consequently, later spikes in a train have smaller amplitude when recorded in the apical dendrites. We found that depolarization and resultant Ca(2+) influx into dendrites caused a persistent facilitation of spike backpropagation. Dendritic patch recordings were made from CA1 pyramidal neurons in mouse hippocampal slices under blockade of fast excitatory and inhibitory synaptic inputs. Trains of 10 backpropagating action potentials induced by antidromic stimulation showed a clear decrement in the amplitude of later spikes when recorded in the middle apical dendrites. After several depolarizing current pulses, the amplitude of later spikes increased persistently, and all spikes in a train became almost equal in size. BAPTA (10 mm) contained in the pipette or low-Ca(2+) perfusing solution abolished this depolarization-induced facilitation, indicating that Ca(2+) influx is required. This facilitation was present in Galpha(q) knock-out mice that lack the previously reported muscarinic receptor-mediated enhancement of spike backpropagation. Therefore, these two forms of facilitation are clearly distinct in their intracellular mechanisms. Intracellular injection of either calmodulin binding domain (100 micrometer) or Ca(2+)/calmodulin-kinase II (CaMKII) inhibitor 281-301 (10 micrometer) blocked the depolarization-induced facilitation. Bath application of a membrane-permeable CaMKII inhibitor KN-93 (10 micrometer) also blocked the facilitation, but KN-92 (10 micrometer), an inactive isomer of KN-93, had no effect. These results suggest that increases in [Ca(2+)](i) cause persistent facilitation of spike backpropagation in the apical dendrite of CA1 pyramidal neuron by CaMKII-dependent mechanisms.

Action Potentials↗
PubMed2000

Critical period for activity-dependent synapse elimination in developing cerebellum.

Synapse elimination is considered to be the final step in neural circuit formation, by causing refinement of redundant connections formed at earlier developmental stages. The developmental loss of climbing fiber innervation from cerebellar Purkinje cells is an example of such synapse elimination. It has been suggested that NMDA receptors are involved in the elimination of climbing fiber synapses. In the present study, we probed the NMDA receptor-dependent period of climbing fiber synapse elimination by using daily intraperitoneal injections of the NMDA receptor antagonist MK-801. We found that blockade of NMDA receptors during postnatal day 15 (P15) and P16, but not before or after this period, resulted in a higher incidence of multiple climbing fiber innervation and caused a mild but persistent loss of motor coordination. Neither basic synaptic functions nor cerebellar morphology were affected by this manipulation. Chronic local application of MK-801 to the cerebellum during P15 and P16 also yielded a higher incidence of multiple climbing fiber innervation. During P15-P16, large NMDA receptor-mediated EPSCs were detected at the mossy fiber-granule cell synapse, but not at the parallel fiber-Purkinje cell or climbing fiber-Purkinje cell synapse. It is therefore likely that the NMDA receptors located at the mossy fiber-granule cell synapse mediate signals leading to the elimination of surplus climbing fibers. These results suggest that an NMDA receptor-dependent phase of climbing fiber synapse elimination lasts 2 d at most. During this phase, the final refinement of climbing fiber synapses occurs, and disruption of this process leads to permanent impairment of cerebellar function.

Aging↗
PubMed2000

mGluR1 in cerebellar Purkinje cells essential for long-term depression, synapse elimination, and motor coordination.

Targeted deletion of metabotropic glutamate receptor-subtype 1 (mGluR1) gene can cause defects in development and function in the cerebellum. We introduced the mGluR1alpha transgene into mGluR1-null mutant [mGluR1 (-/-)] mice with a Purkinje cell (PC)-specific promoter. mGluR1-rescue mice showed normal cerebellar long-term depression and regression of multiple climbing fiber innervation, events significantly impaired in mGluR1 (-/-) mice. The impaired motor coordination was rescued by this transgene, in a dose-dependent manner. We propose that mGluR1 in PCs is a key molecule for normal synapse formation, synaptic plasticity, and motor control in the cerebellum.

Animals↗
PubMed2000

Modest neuropsychological deficits caused by reduced noradrenaline metabolism in mice heterozygous for a mutated tyrosine hydroxylase gene.

Tyrosine hydroxylase (TH) is the initial and rate-limiting enzyme for the biosynthesis of catecholamines that are considered to be involved in a variety of neuropsychiatric functions. Here, we report behavioral and neuropsychological deficits in mice carrying a single mutated allele of the TH gene in which TH activity in tissues is reduced to approximately 40% of the wild-type activity. In the mice heterozygous for the TH mutation, noradrenaline accumulation in brain regions was moderately decreased to 73-80% of the wild-type value. Measurement of extracellular noradrenaline level in the frontal cortex by the microdialysis technique showed a reduction in high K(+)-evoked noradrenaline release in the mutants. The mutant mice displayed impairment in the water-finding task associated with latent learning performance. They also exhibited mild impairment in long-term memory formation in three distinct forms of associative learning, including active avoidance, cued fear conditioning, and conditioned taste aversion. These deficits were restored by the drug-induced stimulation of noradrenergic activity. In contrast, the spatial learning and hippocampal long-term potentiation were normal in the mutants. These results provide genetic evidence that the central noradrenaline system plays an important role in memory formation, particularly in the long-term memory of conditioned learning.

Animals↗
PubMed2000

Relationship between delta-aminolevulinic acid dehydratase genotypes and heme precursors in lead workers.

BACKGROUND: The objective of this study was to investigate the relationships between genotypes of delta-aminolevulinic acid (ALA) dehydratase (ALAD) and disturbances in the heme biosynthetic pathway by lead exposure. METHODS: The subjects were 192 male lead workers and 125 control subjects. Blood lead concentrations (Pb-B), plasma ALA concentrations (ALA-P), and ALAD genotypes were determined for all subjects. In lead workers, ALAD activity, ALA in urine (ALA-U), and erythrocyte zinc protoporphyrin (ZP) were also determined. RESULTS: The frequency of ALAD2 (minor type of ALAD allele) was calculated to be 0.087 in all subjects. No significant relationship was found between ALAD2 frequency and Pb-B levels in lead workers. ALAD1 homozygotes showed significantly higher levels of ZP and ALA-P in comparison with those of ALAD2 carriers at Pb-B levels more than 20 microg/dL and 40 microg/dL, respectively. CONCLUSIONS: ALAD1 homozygotes might be more susceptible than ALAD2 carriers to disturbances in heme metabolism caused by lead exposure.

Adult↗
PubMed2000

The mechanism of a human reaction to vibration stress by palmar sweating in relation to autonomic nerve tone.

OBJECTIVES: To clarify the mechanism of a human reaction to vibration stress by palmar sweating in relation to the autonomic nerve tone. METHODS: The autonomic nerve tone was divided into four types by using digital photoelectroplethysmography (PTG) with auditory stimuli: normal (N), hyperreactive (I and D), and hyporeactive (P) types. Palmar sweating and digital PTG were simultaneously measured on the right palm and middle finger, respectively, in 20 healthy men. The left hand gripping the handle with a grasp strength of 49 N was exposed to vibration at a frequency of 125 Hz and acceleration magnitudes of 0 m/s2 (as a control), 30 m/s2, or 50 m/s2 for 3 min. The volume of palmar sweating was recorded before, during, and 30 min after vibration load. Three kinds of drugs related to the autonomic nervous system were orally administered to the subjects. Then 80 min after administration, the experiments were repeated. RESULTS: Of 20 subjects, 17 showed normal autonomic nerve tone (N type), and 3 hyperreactive (I type). The palmar sweating reaction to vibration in I-type subjects was greater and lasted longer than that in N-type subjects. Vibration with an acceleration of 50 m/s2 produced the greatest reaction which was about 7 times larger than that at 0 m/s2 and 2.5 times that at 30 m/s2 (P < 0.01). Sulpiride decreased palmar sweating during vibration, while prazosin and scopolamine inhibited it. CONCLUSIONS: The palmar sweating reaction to vibration stress was related to the background level of the autonomic nerve tone. The sweating volume was in direct proportion to the acceleration magnitude of vibration. The reaction of palmar sweating to vibration stress may be mediated through both the adrenergic and cholinergic fibers of the autonomic nervous system.

Adrenergic alpha-Agonists↗
PubMed2000

Viability of cryopreserved semilunar valves: an evaluation of cytosolic and mitochondrial activities.

BACKGROUND: Despite long-standing, widespread use of cryopreserved allografts, the basic cellular biology of these tissues is still yet unknown. The present investigation was undertaken to study cryopreserved heart valves from the standpoint of cytosolic esterase and mitochondrial dehydrogenase activities. METHODS: Cryopreserved porcine aortic cusps were observed in an unfixed fresh condition with a confocal laser scanning microscope using fluorescent dye. Porcine cusps and cultured human umbilical vein endothelial cells were divided into three groups, including fresh, cold-preserved, and cryopreserved specimens, and cytosolic esterase activity and mitochondrial dehydrogenase activity were analyzed in each. RESULTS: Confocal laser scanning microscope findings disclosed a widely distributed fluorescence in the cusp. Cytosolic esterase activity within human umbilical vein endothelial cells (28%+/-9.0%) after cryopreservation was significantly less than that it was in the cusps (72%+/-21%). Mitochondrial dehydrogenase activity of cryopreserved human umbilical vein endothelial cells and that of cusps fell to 44%+/-6.1% and 64%+/-17% respectively; the difference between the two values was not significant. CONCLUSIONS: Cryopreservation appeared to produce serious damage to cytosolic and mitochondrial functions of endothelial cells. The cytosolic function of cusps, mainly consisting of fibroblasts, was comparatively preserved after cryopreservation, but mitochondrial function of the cusps was more diminished.

Animals↗
PubMed2000

Heterosynaptic expression of depolarization-induced suppression of inhibition (DSI) in rat hippocampal cultures.

Depolarization-induced suppression of inhibition (DSI) is a transient suppression of the inhibitory synaptic transmission, observed in the hippocampus and the cerebellum, upon postsynaptic depolarization. Using rat hippocampal cultures, we examined whether DSI is confined to the inhibitory synapses on the depolarized neuron or, if DSI can spread to those on neighboring non-depolarized neurons. Whole-cell recordings were performed in 108 neuronal pairs with the following synaptic responses. Stimulation of one neuron evoked the inhibitory autaptic currents (IACs) recurrently in that neuron and also elicited the inhibitory postsynaptic currents (IPSCs) orthodromically in the other neuron. In 38 of 108 pairs, the postsynaptic depolarization caused transient suppression of IPSCs (homosynaptic DSI). In 11 of the 38 pairs exhibiting the homosynaptic DSI, the depolarization also induced suppression of IACs (heterosynaptic DSI). The heterosynaptic DSI, like the homosynaptic DSI, depended on depolarizing pulse duration and was blocked by a phorbol ester. These results suggest that DSI can spread to the synapses on a neighboring non-depolarized neuron in rat hippocampal cultures.

Animals↗
PubMed2000

Local calcium release in dendritic spines required for long-term synaptic depression.

We have used rats and mice with mutations in myosin-Va to evaluate the range and function of IP3-mediated Ca2+ signaling in dendritic spines. In these mutants, the endoplasmic reticulum and its attendant IP3 receptors do not enter the postsynaptic spines of parallel fiber synapses on cerebellar Purkinje cells. Long-term synaptic depression (LTD) is absent at the parallel fiber synapses of the mutants, even though the structure and function of these synapses otherwise appear normal. This loss of LTD is associated with selective changes in IP3-mediated Ca2+ signaling in spines and can be rescued by photolysis of a caged Ca2+ compound. Our results reveal that IP3 must release Ca2+ locally in the dendritic spines to produce LTD and indicate that one function of dendritic spines is to target IP3-mediated Ca2+ release to the proper subcellular domain.

2-Amino-5-phosphonovalerate↗
PubMed2000

Gq protein alpha subunits Galphaq and Galpha11 are localized at postsynaptic extra-junctional membrane of cerebellar Purkinje cells and hippocampal pyramidal cells.

Following cell surface receptor activation, the alpha subunit of the Gq subclass of GTP-binding proteins activates the phosphoinositide signalling pathway. Here we examined the expression and localization of Gq protein alpha subunits in the adult mouse brain by in situ hybridization and immunohistochemistry. Of the four members of the Gq protein alpha subunits, Galphaq and Galpha11 were transcribed predominantly in the brain. The highest transcriptional level of Galphaq was observed in cerebellar Purkinje cells (PCs) and hippocampal pyramidal cells, while that of Galpha11 was noted in hippocampal pyramidal cells. Antibody against the C-terminal peptide common to Galphaq and Galpha11 strongly labelled the cerebellar molecular layer and hippocampal neuropil layers. In these regions, immunogold preferentially labelled the cytoplasmic face of postsynaptic cell membrane of PCs and pyramidal cells. Immunoparticles were distributed along the extra-junctional cell membrane of spines, dendrites and somata, but were almost excluded from the junctional membrane. By double immunofluorescence, Galphaq/Galpha11 was extensively colocalized with metabotropic glutamate receptor mGluR1alpha in dendritic spines of PCs and with mGluR5 in those of hippocampal pyramidal cells. Together with concentrated localization of mGluR1alpha and mGluR5 in a peri-junctional annulus on PC and pyramidal cell synapses (Baude et al. 1993, Neuron, 11, 771-787; Luján et al. 1996, Eur. J. Neurosci., 8, 1488-1500), the present molecular-anatomical findings suggest that peri-junctional stimulation of the group I metabotropic glutamate receptors is mediated by Galphaq and/or Galpha11, leading to the activation of the intracellular effector, phospholipase Cbeta.

Amino Acid Sequence↗
PubMed2000

Formation of hippocampal synapses on patterned substrates of a laminin-derived synthetic peptide.

We created a new culture system which provides simple, stereotyped neuronal circuitries suitable for investigating synaptic events between mammalian central neurons. We used surface chemistry and laser-lithography to produce geometrical patterns of neuron-compatible substrate spaced by less neuron-compatible surfaces. The patterned substrates were composed of a laminin-derived synthetic peptide, PA22-2, and the spacing surfaces of either decyldimethylsilane (DDMS) or trimethylsilane (TMS). Dissociated rat hippocampal neurons survived on the patterned substrates for several days without the aid of glia and extended their neurites along the substrates. The TMS spacing surfaces appeared more favourable for the excitability development and axonal differentiation of the hippocampal neurons, but less favourable for the development of the resting conductance than the DDMS spacing surfaces. Furthermore, neurons grown on the patterned substrates frequently made synaptophysin-positive contacts with one another. Spontaneous post-synaptic currents recorded from such neurons suggest that these contacts were indeed functional synapses. When hippocampal neurons were plated at a very low density, they often formed circuitries consisting of only two neurons on the patterned substrate. Such a simple circuitry allowed us to analyse synaptic transmission in a single neuronal pair without the influence of the third neurons. With the clarity of analysis and the readiness of manipulation, our culture system would offer a powerful tool for studying development and functions of mammalian central synapses.

Animals↗
PubMed2000

Detection of lymph node metastasis of oesophageal cancer by RT-nested PCR for SCC antigen gene mRNA.

With recent development in molecular biology, reverse transcriptase polymerase chain reaction (RT-PCR) has been applied to detect occult lymph node metastasis, but there have been few reports concerning oesophageal cancer. The objective of this study is to investigate the usefulness of the squamous cell carcinoma (SCC) antigen gene as a marker with RT-nested PCR to detect occult lymph node metastases of oesophageal cancer. The SCC antigen has been widely used as a serum tumour marker and was reported as a target gene to detect tumour cells in peripheral blood in cervical cancer. In this study, 620 lymph nodes from 14 oesophageal cancer patients were analysed. The results of RT-nested PCR were compared with that of pathological and immunohistochemical examinations. In the test of sensitivity, the RT-nested PCR detected 10(1) of SCC antigen producing cells in 10(7) peripheral blood mononucleocytes and was not found in 43 control lymph nodes. The pathological examination, immunohistochemical examination and the RT-nested PCR detected 36, 45 and 65 nodes respectively. The RT-nested PCR detected statistically more lymph nodes than the pathological or immunohistochemical examination. The sensitivity and specificity seem higher in squamous cell carcinoma cases. The SCC antigen gene is one of the more useful markers for RT-nested PCR to detect occult lymph node metastases of oesophageal cancer.

Antigens, Neoplasm↗
PubMed2000

Podophyllotoxin aza-analogue, a novel DNA topoisomerase II inhibitor.

The pendant E-ring moiety of the podophyllotoxin aza-analogue 1 that is a potent inhibitor of microtubule assembly was modified in order to acquire inhibitory activity of DNA topoisomerase II. The monophenolic analogue 2 did not exhibit human topoisomerase II inhibition, while the ortho-quinone 3 that was obtained by oxidation of 2 inhibited its catalytic activity (decatenation) in a dose-dependent manner and stimulated double strand DNA breaks in supercoiled circular plasmid DNA, resulting in the production of linear DNA. These results showed that the topoisomerase II inhibition of the ortho-quinone 3 is due to stabilization of the topoisomerase II-DNA covalent binary complex. On the other hand, the ortho-quinone 3 did not inhibit the relaxation process of supercoiled DNA by topoisomerase I at concentrations up to 400 microM, nor was intercalation observed in unwinding measurements of 3. Therefore, the ortho-quinone 3 was shown to be a novel nonintercalative topoisomerase II specific inhibitor that stabilizes the cleavable complex. The present results suggest that the 4'-free hydroxyl group on the E-ring and the sugar moiety on the C-ring are not a prerequisite for topoisomerase II inhibition by podophyllotoxin derivatives.

DNA↗
PubMed2000

Relationship between effects of alcohol on psychomotor performances and blood alcohol concentrations.

Ethanol is a social drug and has been generally known to be a CNS depressant. A large fluctuation of blood alcohol concentration (BAC) is well-known to occur due to main factors such as the genetic polymorphism of the main alcohol metabolizing enzymes and the effect of blood. Few studies have substantially discussed the relationship between impaired CNS activities and BAC. In this study, focusing on the correlation of BAC, we investigated the acute effects of alcohol intake on cognitive performance in humans by objective evaluation methods consisting of the attention-demanding cognitive tasks. Tasks were administered to ten healthy male volunteers before and after ingesting established amounts of alcohol. With increased BAC, we observed prolongation of reaction time performances and lowering of a coordination performance. From the results, we concluded that cognitive performance deteriorates with an increase of BAC. Additionally, the BAC threshold that causes significant impairment of cognitive performance was estimated to be approximately 50 mg/dl (ca. 10 mM).

Adult↗