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K Hama

Publications and source records attributed to K Hama.

At least 73 records · Page 4Linked to original sources

Catecholaminergic neurons containing GABA-like and/or glutamic acid decarboxylase-like immunoreactivities in various brain regions of the rat.

The coexistence of immunoreactivities for tyrosine hydroxylase (TH) and glutamic acid decarboxylase (GAD) and/or gamma-aminobutyric acid (GABA) was revealed in various brain regions in colchicine-injected and untreated rats, using the peroxidase-antiperoxidase method. Consecutive 40 micron thick Vibratome sections were incubated in different antisera and those cells which were bisected by the plane of sectioning so as to be included at the paired surfaces of two adjacent sections were identified. The coexistence of the immunoreactivities for TH and GAD or GABA in the same cell could thus be determined by observing the immunoreactivity of the two halves of the cell incubated in two different antisera. In the olfactory bulb, retina, diencephalon, mesencephalic central grey and cerebral cortex, many TH-like immunoreactive neurons also showed GAD-like or GABA-like immunoreactivity, whereas in the substantia nigra, ventral tegmental area and locus ceruleus none of TH-like immunoreactive neurons showed either GAD-like or GABA-like immunoreactivity. In the olfactory bulb, retina and cerebral cortex, the majority of the TH-like immunoreactive neurons were also GAD-like or GABA-like immunoreactive. In the diencephalon of colchicine-injected rats, at least one-third of the TH-like immunoreactive neurons were GAD-like immunoreactive. Using serial 0.5 micron thick plastic-embedded sections, it was shown that immunoreactivities for three antigens, GAD, GABA and TH could occur in the same neurons in the olfactory bulb. These observations indicate the possible coexistence of two classical transmitters. GABA and catecholamine, in various brain regions of the rat.

Animals↗

Tyrosine hydroxylase-immunoreactive intrinsic neurons in the rat cerebral cortex.

Using specific antisera against the catecholamine synthesizing enzyme, tyrosine hydroxylase (TH), in combination with the peroxidase-antiperoxidase method and/or the avidin-biotin complex method, we have found a new group of TH immunoreactive (TH-I) neurons in the rat cerebral cortex. Numerous TH-I cells were observed all over the isocortex, that is, frontal, temporal, parietal and occipital regions, and in some parts of the allocortex such as the anterior cingulate cortex, the retrosplenial cortex and anterior part of the insular cortex. In contrast, they were rare in the perirhinal cortex, posterior part of the insular cortex, piriform cortex, entorhinal cortex and hippocampal formation. TH-I cells were situated throughout all cortical layers, but were most concentrated in layer II/III. Although TH-I cells were heterogeneous in shape, the majority were bipolar. All TH-I cells so far examined appeared to be of the nonpyramidal type. The majority of these intrinsic TH-I neurons also contained the GABA-like immunoreactivity and thus could be regarded as a subpopulation of cortical GABAergic neurons.

Animals↗

Intramembrane particles and terminal responses following denervation of frog muscle spindles.

The sizes and population density of intramembrane particles in the non-myelinated sensory nerve terminals of frog muscle spindles were studied in comparison with changes in the responses to spindles to stretch 1-28 days after denervation. The sciatic nerve in the right leg was cut at the dorsal part of the hip joint. Spindle receptors in the left leg were used as controls. Muscle spindles were isolated from semitendinosus and iliofibularis muscles. The mean density of particles, which occurred mainly on the protoplasmic (P) face, declined rapidly to 60% of the control values 2 days after denervation, 30% after 4 days and to less than 10% after 7 days. The mean diameter of particles changed from 8.0 +/- 1.5 nm in the control to 12.8 +/- 2.5 nm 7 days after denervation. No significant difference between the results for the trunks (greater than 2 micron in diameter) and the branches (less than 0.5 micron) of the non-myelinated axons was observed. The peak rate of afferent discharges during a ramp-and-hold stretch survived up to 4 days after denervation, whereas the static component disappeared after 3 days. It is suggested that the decline to less than 20% of the control density of particles removes the mechanosensitivity of the sensory endings.

Animals↗

Three-dimensional structure of astrocytes in the rat dentate gyrus.

Structural features of astrocytes in the rat dentate gyrus were studied by means of light and high-voltage electron microscopy of Golgi-impregnated materials, conventional electron microscopy, combined Golgi-electron microscopy, and immunohistochemistry for glial fibrillar acidic protein. Astrocytes in the dentate gyrus were of the protoplasmic type and were classified into six subtypes based on the location of their somata; i.e., astrocytes in the polymorph layer, in the subgranular zone, in the granular cell layer, at the border of the granular cell layer and the molecular layer, in the molecular layer, and subjacent to the pia surface. Stereoscopic observations of 5-micron-thick sections of Golgi-impregnated materials revealed three-dimensional structural details of astroglial processes not apparent in either the light microscope or in conventional thin-section electron microscopy. Most of them were basically thin sheets, varying in shape and size in accordance with their sites. In the granule cell layer, thin veillike sheets or lamellae, originating from three kinds of astrocytes (subtypes 2, 3, and 4), intervened between granule cell somata, whereas in the plexiform and molecular layers small leafletlike appendages originating from astrocytes (subtypes 1, 2, 4, and 5) intermingled with one another, making spongelike conglomerates. Thus astrocytes in the subgranular zone and those at the border of the granule cell layer and the molecular layer showed prominent regional differentiation of their processes among layers. In addition to these sheetlike processes, thin threadlike processes were also common.

Animals↗

The transverse tubular system of rat myocardium: its morphology and morphometry in the developing and adult animal.

The three dimensional arrangements of the T system in the developing and adult animal were investigated by means of high voltage electron microscope stereoscopy using Golgi treated materials. The rat myocardial T system was composed of three major group elements: the transverse tubules, longitudinal tubules and flattened cisternae, which were classified according to their orientation and to their morphological features. It was found, as the growth of the rats proceeded, that the longitudinal tubules increased in number and that the transverse tubules were arranged more regularly and densely at the level of the z band. The flattened cisternae transiently increased in number during the 2-9 weeks, and then decreased gradually. Electron microscopy also revealed that all the transverse, longitudinal tubules and flattened cisternae of the T system had the chance of forming a coupling with the sarcoplasmic reticulum irrespective of its morphology and orientation to the myofibrils. Quantitative analysis of the rat T system from the stereo images indicated that the surface area (0.299 micron2/micron3) was considerably greater than previously reported.

Age Factors↗

Use of high concentrations of glutaraldehyde for immunocytochemistry of transmitter-synthesizing enzymes in the central nervous system.

Aldehyde fixatives containing high concentrations of glutaraldehyde, usually used for conventional electron microscope studies, were successfully used for immunocytochemistry of transmitter synthesizing enzymes, glutamate decarboxylase and tyrosine hydroxylase, in the rat central nervous system. Although a high concentration of glutaraldehyde could cause tremendous non-specific staining, this was almost completely absent after treating sections with 1% sodium borohydride for 30 min. Furthermore, it was shown that a high concentration of glutaraldehyde might cause no appreciable reduction of the antigenicities of glutamate decarboxylase and tyrosine hydroxylase when compared with fixatives containing a low concentration of glutaraldehyde. It is suggested that fixatives containing high concentrations of glutaraldehyde are very useful, not only for conventional electron microscope studies, but also for light and electron microscope immunocytochemistry of some antigens, including glutamate decarboxylase and tyrosine hydroxylase.

Aldehydes↗

Detection of delta 9-THC in saliva by capillary GC/ECD after marihuana smoking.

A method is described for the determination of delta 9-tetrahydrocannabinol (delta 9-THC) in the saliva by the use of a combination of moving-precolumn injector and glass capillary gas chromatograph with electron capture detector (GC/ECD). There were no interfering peaks due to impurities around the peak of pentafluoropropyl derivative of delta 9-THC (delta 9-THC-PFP). This GC/ECD method was linear over the range of 5-200 ng/ml of delta 9-THC-PFP. The lower detection limit was approximately 1 ng/ml. delta 9-THC content in the saliva after experimental marihuana smoking was measured by this method. It was demonstrated that for at least 4 h after smoking the level of delta 9-THC was sufficient for detection.

Adult↗

Observations on the fine structure of nodal, Purkinje and working myocardial cells isolated from rabbit hearts.

Sinoatrial and atrioventricular nodal cells as well as Purkinje and working myocardial cells isolated enzymatically from rabbit hearts were examined by transmission electron microscopy. The cardiac cell fine structure remained well organized in the storage solution after isolation. In normal Tyrode solution, the nodal cells changed from spindle shaped to spherical, whereas the shapes of Purkinje and working myocardial cells remained unchanged. The nodal cell fine structure became disorganized with respect to its sarcomeric arrangement in normal Tyrode solution. This was assumed to result from a combination of several factors seen in the nodal cells: the loss of the normal anchoring of the myofibrils at the previous intercalated discs, breakdown of the Z bands and the alteration of the integrity of the intermediate filaments. Also, extensive restoration of plasma membrane damaged by of the intermediate filaments. Also, extensive restoration of plasma membrane damaged by the isolation procedure was observed in this solution. This might correspond to the retention of the normal physiological properties of the plasma membrane of the nodal cells despite gross morphological changes undergone in normal Tyrode solution.

Animals↗

Tetrahydro-beta-carbolines in human urine and rat brain--no evidence of formation by alcohol drinking.

1-Methyl-1,2,3,4-tetrahydro-beta-carboline (1-MeTHBC) and 6-methoxy-1-methyl-1,2,3,4-tetrahydro-beta-carboline (6-M-1-MeTHBC) were identified and quantified in the urine of 131 healthy volunteers by gas chromatography/mass spectrometry (GC/MS) with a chemically-bonded glass-capillary column. 1-MeTHBC was present naturally in the human urine, whereas 6-M-1-MeTHBC was observed in only 20% of all subjects at a much lower level. No significant relationship was observed between these substances and alcohol consumption in the subjects. Various foods and alcoholic beverages were found to contain 1-MeTHBC but not 6-M-1-MeTHBC; thus, the urinary 1-MeTHBC detected could be of dietary origin. Salsolinol, on the other hand, was significantly elevated in the urine without increase of THBCs by long-term moderate drinking. 1-MeTHBC and 6-M-1-MeTHBC were also analysed in the brains of rats following inhalation of ethanol and acetaldehyde for four days. No evidence of formation of these compounds was observed in any regions of the brain.

Acetaldehyde↗

GABAergic neurons containing CCK-8-like and/or VIP-like immunoreactivities in the rat hippocampus and dentate gyrus.

The coexistence of cholecystokinin-octapeptide-like (CCK-L) and/or vasoactive-intestinal-polypeptide-like immunoreactive (VIP-LI) materials and glutamate decarboxylase (GAD) was studied in the rat hippocampus and dentate gyrus by means of immunohistochemistry. Consecutive 40-micron-thick sections were incubated in different antisera and those cells which were bisected by the plane of sectioning so as to be included at the paired surfaces of two adjacent sections were identified. The coexistence of the immunoreactivities for these peptides and GAD in the same cell could thus be determined by observing the immunoreactivity of the two halves of the cell, incubated in two different antisera. Almost all of the CCK-LI neurons were also GAD immunoreactive, whereas only about 10% of the GAD-immunoreactive neurons were CCK-LI. The percentages of GAD-immunoreactive neurons which were also immunoreactive for CCK were dependent on the laminar area in which they were found: i.e., 15-20% in the stratum radiatum of the hippocampus, about 10% in the stratum pyramidale, and about 6% in the stratum oriens. In contrast to the CCK-LI neurons, only about 40% of the VIP-LI neurons were identified to be also GAD immunoreactive, which might correspond to only part of the GAD-immunoreactive neurons. Furthermore the coexistence of VIP-LI and CCK-LI materials was recognized in about 10% of the CCK-LI neurons or about 35% of the VIP-LI neurons, indicating that some GABAergic neurons (presumably about 1%) in the rat hippocampus and dentate gyrus may contain both CCK-LI and VIP-LI materials.

Animals↗

Coexistence of immunoreactivities for glutamate decarboxylase and tyrosine hydroxylase in some neurons in the periglomerular region of the rat main olfactory bulb: possible coexistence of gamma-aminobutyric acid (GABA) and dopamine.

The coexistence of immunoreactivities for glutamate decarboxylase (GAD) and tyrosine hydroxylase (TH) was revealed in some neurons in the periglomerular region and in the superficial part of the external plexiform layer of the rat main olfactory bulb. In neurons showing the immunoreactivity for either GAD or TH, about 10-55% showed both immunoreactivities.

Animals↗

Gap junctions between non-pyramidal cell dendrites in the rat hippocampus (CA1 and CA3 regions): a combined Golgi-electron microscopy study.

Non-pyramidal cells in the rat hippocampus were examined with a combined Golgi-electron microscopic method. The somata of non-pyramidal cells were ovoid, about 15 X 30 micron, and several smooth and/or varicose dendrites extended from them. With electron microscopy, Golgi-impregnated gold-toned non-pyramidal cells showed distinctive fine structural features. The somata displayed large nuclei and an extensive perikaryal cytoplasm. The nuclei showed extensive cytoplasmic invaginations, little heterochromatin, conspicuous nucleoli, and intranuclear rods composed of filamentous bundles. The perikaryal cytoplasm was rich in cell organelles such as well-developed cisternae of rough endoplasmic reticulum and Golgi apparatus, and numerous clusters of free ribosomes and mitochondria. Many synaptic boutons, most of which formed asymmetrical synapses, impinged upon the somata and dendrites. Gap junctions were seen on varicose dendrites of Golgi-impregnated non-pyramidal cells. These gap junctions were patch-like, about 0.1-0.6 micron in diameter, and situated in the stratum radiatum or stratum oriens of the CA1 and CA3 regions 70-230 micron from the soma. They displayed a characteristic cytoplasmic semidense material undercoating the junctional membranes. The gap junctions were usually formed between impregnated and unimpregnated varicose dendrites. Thirteen of a total of 22 gap junctions involving the impregnated dendrites were situated singly, whereas the remaining nine were on four impregnated dendrites in clusters of two or three side by side. In the latter cases, two pairs of junctions were formed between pairs of dendrites running parallel to each other, and each of the other two pairs was formed among three dendrites, appearing to make a dendritic network bridged by gap junctions. One gap junction was seen between two impregnated dendrites originating from two identified Golgi-impregnated non-pyramidal cells. These observations revealed unequivocally that non-pyramidal cells in the hippocampus form gap junctions with one another on their dendrites.

Animals↗

Light and electron microscopy study of fusion of facial prominences. A distinctive type of superficial cells at the contact sites.

The contact site between the medial nasal prominence (MNP) and the lateral nasal prominence (LNP) during the period of primary palate formation in the mouse embryo was examined by light and electron microscopy. Throughout this period, a distinctive type of superficial cell was observed at the contact site. These superficial cells had a large nucleus and abundant cytoplasm as well as structural features characteristic of embryonic cells. At earlier stages, these cells were seen at the transitional region between the surface ectoderm and the epithelia of the nasal pit at the end of the isthmus, where initial contact of opposing MNP and LNP took place. At later stages, these superficial cells appeared to bridge the gap between MNP and LNP at the contact sites, which extended to the bottom of the valley formed by MNP and LNP. These cells were also observed on the surface near the contact sites, that is, the presumptive fusion area. These superficial cells displayed well-developed junctional complexes (intermediate and gap junctions, and desmosomes). Many filaments were observed subjacent to the plasma membranes of these superficial cells, some of which were associated with junctional complexes. These observations suggest that this kind of distinctive superficial cell may play critical roles in the contact of MNP and LNP throughout the fusion process.

Animals↗

A study of the myenteric plexus of the congenital aganglionosis rat (spotting lethal).

The entire bowel of a mutant strain of rats, the congenital aganglionosis rat (spotting lethal), was investigated using the acetylcholinesterase reaction and immunohistochemical staining for tyrosine hydroxylase and substance P in whole-mount preparations. The histology of the bowel of mutant rats was also studied by light- and electron microscopy. In all examined mutant rats, a constricted region of intestine followed a dilated region of the bowel. In 29 cases constricted segments extended from rectum to distal ileum; in 3 cases from rectum to middle colon. In controls the myenteric plexus appeared as a mesh-work consisting of ganglion strands and internodal strands, showing a rather regular ladder-like pattern from duodenum to rectum. The myenteric plexus of mutants was very different from that of controls, showing conspicuous regional differences. Even in the duodenum, where there was no macroscopical disorder, the plexus showed an irregular pattern, the meshes varying greatly in size and shape. Ganglion strands were shorter than those in controls. The plexus in the dilated segment gradually decreased in density, finally disappearing above the proximal terminal of the constricted segment. In some areas below this transition, i.e., the anal portion of the constricted segment in 29 cases (long constricted segment type only), there were neither ganglion cells nor nerve fibers except for scarcely distributed tyrosine hydroxylase-immunoreactive nerve fibers. In the distal part of the upper colon some fine nerve bundles appeared to run irregularly. These nerve bundles gradually increased in number and mixed with thicker nerve bundles in the lower portion of the colon. Finally, at the level of the rectum, nerve bundles of various sizes interlaced irregularly with one another to form a network. However, this network was free from ganglion cells.

Acetylcholinesterase↗