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Biomedical subjects

Y Shinoda

Publications and source records attributed to Y Shinoda.

At least 73 records · Page 4Linked to original sources

Axon collaterals of mossy fibers from the pontine nucleus in the cerebellar dentate nucleus.

1. Single axons of pontine nucleus neurons (PN axons) receiving cerebral input were stained intra-axonally with horseradish peroxidase (HRP) in the cerebellum of cats. The axonal trajectory of single PN axons was reconstructed from serial sections of the cerebellum and the brain stem. 2. Axons were penetrated in the white matter near the dentate nucleus, and, after electrophysiological identification, PN axons were injected iontophoretically with HRP. The identification criteria for the PN axons were 1) their direct responses to stimulation of the contralateral pontine nucleus (PN), 2) their synaptic activation from the contralateral cerebral cortex, and 3) the decrease in threshold for evoking direct spikes in stimulation of the PN by conditioning stimuli applied in the cerebral cortex. 3. Two hundred thirty-three axons were electrophysiologically identified as PN axons receiving the input from the cerebral cortex. Ninety-six of them were stained successfully with HRP, and reconstructions were made from 40 well-stained PN axons. All of them gave rise to mossy fibers and terminated in the granular layer of the cerebellar cortex as typical mossy fiber rosettes. Out of these, 22 gave axon collaterals to the dentate nucleus. Virtually all of the axon branches observed in the dentate nucleus were axon collaterals of mossy fibers from the PN to the cerebellar cortex. In 7 of these 22 PN axons, cell bodies were retrogradely labeled with HRP, and all of them were found in the contralateral PN. 4. The stained-stem axons arising from the PN ran medially in the pons, crossed the midline, and then ascended dorsocaudally in the branchium pontis. After passing in the white matter anterior to or lateral to the dentate nucleus, they entered into the cerebellar cortex. On their way, one to three axon collaterals were given off from parent axons to the dentate nucleus. The diameter of these collaterals was very thin (mean, 0.6 microns), compared with the large diameter of the parent axons (mean, 2.1 microns). 5. Some axon collaterals were very simple and had only one terminal branch with or without short branchlets, whereas others were more complex, and single axon collaterals ramified before forming a terminal arborization. Axon collaterals of single PN axons mainly spread mediolaterally or dorsoventrally in the frontal plane but had a very narrow rostrocaudal extension. 6. Terminal branches usually bore swellings en passant along their length and one terminal swelling at their end. The number of swellings per axon collateral ranged 23-180 (116 +/- 52, mean +/- SD).(ABSTRACT TRUNCATED AT 400 WORDS)

Afferent Pathways↗

[Effects of oxiracetam on the decrease in population spikes in hypoxic and low glucose media].

Evoked potentials were recorded in rat cerebral cortical slices. The amplitude of the evoked potential was reduced by perfusion with hypoxic (0-25%) or low glucose (0-5 mM) media in a concentration-dependent manner, and the evoked potentials disappeared under severe conditions (below 15% O2, below 3 mM glucose). We investigated the protective effects of oxiracetam on the decrease in evoked potentials under hypoxic (15% O2) and low glucose (3 mM glucose) conditions. Drugs were perfused from 45 min before hypoxic or low glucose perfusion to the end of the experiment. Oxiracetam (10(-6)-10(-5) M) dose-dependently minimized the amplitude reduction of evoked potentials and prolonged their disappearance time. At a concentration of 10(-5) M, oxiracetam protected against the disappearance of evoked potential in 5 of the 6 samples under hypoxic conditions and in all 6 samples under low glucose conditions. Indeloxazine (5 x 10(-6)-10(-5) M) and bifemelane (5 x 10(-6)-10(-5) M) prevented the reduction of the amplitude of evoked potentials under low glucose conditions. However, these drugs had no effect at a concentration of 10(-6) M. These data indicate that oxiracetam has a protective effect against neuronal dysfunction and that this effect develops at a lower concentration than those of indeloxazine and bifemelane.

Animals↗

[Biochemical studies of oxiracetam (CT-848) on cholinergic neurons].

Effects of oxiracetam on cholinergic neurons were investigated by biochemical methods. 1) Oxiracetam did not inhibit 3H-QNB binding in the cerebral cortex and hippocampus. 2) In the 3H-QNB binding study, oxiracetam did not change the inhibition-concentration curve for the muscarinic agonist carbachol and had no effect on GppNHp-induced inhibition of oxotremorine binding. 3) Oxiracetam (10-100 microM) enhanced K(+)-induced ACh release from slices of rat hippocampus. 4) In the in vitro perfusion studies, oxiracetam (10-100 microM), but not aniracetam and piracetam, enhanced choline-acetyltransferase (ChAT) activity in the hippocampal slices. 5) Repeated administration of oxiracetam (100 or 500 mg/kg, p.o., once daily) to old rats significantly enhanced ChAT activities in the cerebral cortex, hippocampus and striatum, while it did not influence the Bmax and Kd for 3H-QNB binding in the hippocampus. 6) Oxiracetam did not affect the acetylcholinesterase activity in mouse brain homogenate. These results suggest that oxiracetam enhances precholinergic functions.

Acetylcholine↗

The effects of oxiracetam (CT-848) on local cerebral glucose utilization after focal cerebral ischemia in rats.

The effects of oxiracetam on the reduction of brain metabolism induced by focal cerebral ischemia were investigated by measuring local cerebral glucose utilization (LCGU) in rats 24 hr after left middle cerebral artery occlusion. Focal cerebral ischemia reduced LCGU in the entire ipsilateral cortex, the greatest reduction being in the lateral parts of the frontoparietal cortex. LCGU was slightly reduced in the contralateral cortex; this reduction was considered to be caused by diaschisis. Oxiracetam was administered intraperitoneally for 3 days prior to middle cerebral artery occlusion. In the ipsilateral cortex, LCGU reduction was minimized in the ischemic center areas by oxiracetam at a dose of 400 mg/kg and in more extensive areas, by a dose of 800 mg/kg. Moreover, oxiracetam at a dose of 800 mg/kg enhanced metabolism impaired by diaschisis in the caudal areas of the contralateral cortex. These findings suggest that oxiracetam minimizes the reduction of brain function induced by ischemia and may therefore be useful in the treatment of cerebrovascular disease.

Animals↗

Immunohistochemical localization of type IV collagen and laminin in the gingival capillary basement membrane of the diabetic rat.

Distribution of type IV collagen and laminin in the gingival capillary basement membrane from streptozotocin-induced diabetic rats was investigated using immunoelectron microscopy. Both type IV collagen and laminin were found throughout the basement membrane. Quantitative analysis revealed that the immunoreactive area for laminin did not change with age, and the width of laminin deposition remained constant, even when diabetes was induced in the animals. However, the immunoreactive area for type IV collagen thickened with age. Further, the width of type IV collagen in the basement membrane increased markedly 36 weeks after diabetes was induced. It was concluded that the thickening of the gingival capillary basement membrane in experimentally induced diabetic rats was due an increase of type IV collagen deposition.

Age Factors↗

Appearance of NMDA receptors triggered by anoxia independent of voltage in vivo and in vitro.

Using rat hippocampus we have studied the pattern of neuronal death, abnormal discharge and loss of electrical excitability in slices prepared from animals subjected to bilateral, four-vessel cerebral anoxia and in slices prepared from normal animals that are subjected to anoxia in the recording chamber. As others have reported, pyramidal neurons in area CA1 are lost first after anoxia, while CA3 neurons have an intermediate sensitivity, and those in dentate are relatively anoxia-resistant. After anoxic damage to the intact animal, neurons in both CA1 and CA3 show abnormal bursting discharges in response to synaptic activation for several days, and then the response in CA1 decreases in amplitude and finally the area become unexcitable. While antagonists for N-methyl-D-aspartate (NMDA) receptors have essentially no effect on synaptic responses in control animals, they reduce the bursting responses and greatly depress the small responses in CA1 as neurons are becoming unexcitable after anoxia. With intracellular recording CA1 neurons from animals made transiently anoxic, in contrast to controls, show prolonged synaptic responses, the later components of which are blocked by NMDA antagonists. When slices from normal animals are subjected to anoxia such that excitability is totally lost over a period of about 10 min, there is no significant membrane depolarization during the anoxic episode and recovery of excitability occurs with reoxygenation. However, a period of hyperexcitability and bursting follows and electrical excitability is lost in CA1 but not CA3 neurons after about 90 min.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Monoclonal antibodies toward scallop (Patinopecten yessoensis) testis and wheat germ calmodulins.

A monoclonal antibody (IM7) toward scallop testis calmodulin and another one (PBE2) toward wheat germ calmodulin were produced. Ca2+ was required for IM7 to react with scallop calmodulin. IM7 reacted with the C-terminal region (Asp78-Lys148) of the calmodulin. As observed on competitive ELISA, IM7 reacted with chicken calmodulin, but not with Euglena gracilis or wheat calmodulin, troponin C, myosin light chains, or parvalbumin. It is assumed that the cluster of Thr143, Thr146, and Ser147 in the C-terminal region acts as the antigenic site. IM7 (and Fab of IM7) inhibited the activities of myosin light chain kinase and cAMP-phosphodiesterase. PBE2 reacted with wheat germ calmodulin irrespective of the presence or absence of Ca2+, the antigenic site being in the N-terminal region (Ala1-Met37). It reacted with wheat and spinach calmodulins, but not with scallop, chicken, or Euglena calmodulin, troponin C, myosin light chains, or parvalbumin. PBE2 had no effect on the activities of myosin light chain kinase and cAMP-phosphodiesterase.

3',5'-Cyclic-AMP Phosphodiesterases↗

Parietal projection of thalamocortical fibers from the ventroanterior-ventrolateral complex of the cat thalamus.

Anterograde labelling following focal injections of Phaseolus vulgaris leucoagglutinin was used to identify the parietal distribution of thalamocortical (TC) fibers from the ventroanterior-ventrolateral (VA-VL) complex of the cat thalamus. In injections in the ventrolateral or the caudal part of the VA-VL complex, labelled TC fibers were distributed in layers I, III and IV of the parietal areas 5a and/or 5b, whereas in injections located more rostrally or dorsomedially, labelled TC fibers were almost confined to layer I.

Animals↗

Preliminary crystallographic study of phospholipase A2 from the venom of Trimeresurus flavoviridis (habu snake).

Crystallization and a preliminary crystallographic study of Trimeresurus flavoviridis (habu snake) phospholipase A2 (PLA2) were carried out. Although crystals were obtained from various solutions, crystals suitable for X-ray analysis could be obtained from polyethylene glycol solutions only when a repeated seeding technique was applied starting from twinned crystals. The crystal is monoclinic with space group P21, with a = 44.1, b = 55.7, c = 48.8 A, and beta = 92.4 degrees. An asymmetric unit contains a dimer consisting of two identical subunits made of 122 amino acids. The crystal reflects X-rays beyond 2.5 A. A Pt derivative gave a good isomorphous crystal.

Animals↗

Distribution of terminals of thalamocortical fibers originating from the ventrolateral nucleus of the cat thalamus.

Anterograde labelling following focal injections of Phaseolus vulgaris leucoagglutinin was used to identify the threedimensional cortical distribution of thalamocortical (TC) fibers from the ventrolateral nucleus of the thalamus of the cat. The labelled TC fibers were distributed usually in layers I and III of the motor cortex and the terminals in layer III tended to aggregate into patches about 1-1.5 mm wide in a mediolateral direction. These patches were arranged in longitudinal strips about 2-5 mm long in a rostrocaudal direction and were separated by gaps of terminal free area.

Animals↗

Comparison of the branching patterns of lateral and medial vestibulospinal tract axons in the cervical spinal cord.

The morphology of single physiologically-identified lateral and medial vestibulospinal tract (LVST and MVST) axons was analysed, using intracellular staining with horseradish peroxidase (HRP) and three-dimensional reconstruction of axonal trajectories in the cat. Axons were penetrated in the cervical cord at C1-C8 with a microelectrode filled with 7% HRP. These axons were identified as vestibulospinal axons by their monosynaptic responses to stimulation of the vestibular nerve and further classified as either LVST or MVST axons by their responses to stimulation of the LVST and MVST. The stained axons could be traced over distances of 3-16 mm rostrocaudally. Within these lengths, both LVST and MVST axons were found to have multiple axon collaterals at different segments in the cervical cord. Up to seven collaterals were given off from the stems of MVST axons and LVST axons. The LVST axons included both neurones terminating at the cervical cord and those projecting further caudally to the thoracic or lumbar cord. Each collateral of these LVST axons, after entering into the gray matter, ramified successively in a delta-like fashion and terminated mainly in lamina VIII and in the medial part of lamina VII. Many boutons of both terminal and en passant types seemed to make contact with the cell bodies and proximal dendrites of neurones in the ventromedial nucleus (VM). Each collateral had a narrow rostrocaudal extension (0.2-1.6 mm, average 0.8 mm) in the gray matter in contrast to a much wider intercollateral interval (average 1.5 mm), so that there were gaps free from terminal boutons between adjacent collateral arborizations. The morphology of axon collaterals of MVST axons was very similar to that of LVST axons. The rostrocaudal extent of single axon collaterals was very restricted (0.3-2.1 mm) in contrast to the wide spread in a mediolateral or a dorsoventral direction. MVST axons had intensive projections to the upper cervical cord with multiple axon collaterals. One to seven collaterals of single MVST axons were found at C1-C3. Terminals of MVST axons were distributed in laminae VII, VIII and IX, including the VM, the nucleus spinalis n. accessorii (SA), and the commissural nucleus. Many terminals seemed to make contact with retrogradely-labelled motoneurones of neck muscles. Both axosomatic and axodendritic contacts were observed on motoneurones in various sizes. Some collaterals gave rise to terminal arborizations in both the VM and the SA. These results suggest that single LVST and MVST axons may control excitability of multiple dorsal axial muscles concurrently with their multiple axon collaterals at multisegmental levels.

Animals↗

Identification of troponin-I of crayfish myofibrils.

Crayfish (Procambarus clarkii) myofibrils contain two basic proteins of molecular weights of 25,000 and 23,000. Both of the two proteins inhibit actomyosin ATPase as the vertebrate troponin-I does. These results differ from the previous one that troponin-I of crayfish (Astacus leptodactylus) showed a single band on polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAGE).

Adenosine Triphosphatases↗

Excitatory inputs to cerebellar dentate nucleus neurons from the cerebral cortex in the cat.

1. In anesthetized cats, we investigated excitatory and inhibitory inputs from the cerebral cortex to dentate nucleus neurons (DNNs) and determined the pathways responsible for mediating these inputs to DNNs. 2. Intracellular recordings were made from 201 DNNs whose locations were histologically determined. These neurons were identified as efferent DNNs by their antidromic responses to stimulation of the contralateral red nucleus (RN). Stimulation of the contralateral pericruciate cortex produced excitatory postsynaptic potentials (EPSPs) followed by long-lasting inhibitory postsynaptic potentials (IPSPs) in DNNs. The most effective stimulating sites for inducing these responses were observed in the medial portion (area 6) and its adjacent middle portion (area 4) of the precruciate gyrus. Convergence of cerebral inputs from area 4 and area 6 to single DNNs was rare. 3. To determine the precerebellar nuclei responsible for mediation of the cerebral inputs to the dentate nucleus (DN), we examined the effects of stimulation of the pontine nucleus (PN), the nucleus reticularis tegmenti pontis (NRTP) and the inferior olive (IO). Systematic mapping was made in the NRTP and the PN to find effective low-threshold stimulating sites for evoking monosynaptic EPSPs in DNNs. Stimulation of either the PN or the NRTP produced monosynaptic EPSPs and polysynaptic IPSPs in DNNs. Using a conditioning-testing paradigm (a conditioning stimulus to the cerebral peduncle (CP) and a test stimulus to the PN or the NRTP) and intracellular recordings from DNNs, we tested cerebral effects on neurons in the PN and the NRTP making a monosynaptic connection with DNNs. Conditioning stimulation of the CP facilitated PN- and NRTP-induced monosynaptic EPSPs in DNNs. This spatial facilitation indicated that the excitatory inputs from the cerebral cortex to DNNs are at least partly relayed via the PN and the NRTP. 4. Stimulation of the contralateral IO produced monosynaptic EPSPs and polysynaptic IPSPs in DNNs. These monosynaptic EPSPs were facilitated by conditioning stimulation of the CP, strongly suggesting that the IO is partly responsible for mediating excitatory inputs from the cerebral cortex to the DN. A comparison was made between the latencies of IO-evoked IPSPs in DNNs and the latencies of IO-evoked complex spikes in Purkinje cells. Such a comparison indicated that the shortest-latency IPSPs evoked from the IO were not mediated via the Purkinje cells and suggested the pathway mediated by inhibitory interneurons in the DN.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

[The neuromuscular blocking activity of isepamicin sulfate (HAPA-B) compared with other aminoglycoside antibiotics].

Effects of isepamicin sulfate (HAPA-B), a new aminoglycoside antibiotic, on the neuromuscular transmission were studied in rats and compared with those of amikacin (AMK) or other aminoglycoside antibiotics. The HAPA-B, as well as other aminoglycoside antibiotics, depressed the twitch response of diaphragm to phrenic nerve stimulation in vitro. The depression effects of different drugs were compared and graded in the order of strengths of blocking action as: netilmicin (NTL) greater than gentamicin (GM) greater than streptomycin (SM) greater than kanamycin (KM) greater than AMK greater than HAPA-B. The IC50 (concentration which inhibited the response by 50%) of HAPA-B was 3.6 X 10(-3) g/ml. The neuromuscular blockade produced by HAPA-B was reversed by CaCl2, KCl or caffeine but not by neostigmine. D-Tubocurarine or MgCl2 augmented the neuromuscular effects of HAPA-B. Intramuscular (400 mg/kg) and intravenous (100 mg/kg) injections of HAPA-B did not affect the twitch response of gastrocnemius muscle to sciatic nerve stimulation in situ. Intravenous injection of 200 mg/kg caused death in some rats and depression of the twitch response in others. Intravenous AMK produced no significant effect on the twitch response at 50 mg/kg and caused death at 100 mg/kg. GM and SM caused death or significant degree of depression of the twitch response at intravenous doses of 50 mg/kg. In experiments of intravenous drug infusion for 60 minutes, the twitch response was depressed by HAPA-B at 400 mg/kg/hr and by AMK at 200 and 400 mg/kg/hr. In conclusion, HAPA-B has a neuromuscular blocking action presumably at the nerve terminal. However, its action was the weakest among the aminoglycoside antibiotics tested.

Amikacin↗