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Biomedical subjects

K Sugaya

Publications and source records attributed to K Sugaya.

At least 91 records · Page 5Linked to original sources
PubMed1995

Inhibitory influence from the nucleus reticularis pontis oralis on the micturition reflex induced by electrical stimulation of the pontine micturition center in cats.

An inhibitory influence from the nucleus reticularis pontis oralis on the micturition induced by electrical stimulation of the pontine micturition center was investigated in decerebrate cats at the supracollicular level. Monopolar electrical stimulation (30-50 microA, 0.2 ms, 50 Hz, 3 s) of the nucleus locus coeruleus alpha (pontine micturition center) produced micturition similar to that induced by distending the bladder by saline infusion. The intravesical pressure increase and micturition induced by electrical stimulation of the nucleus locus coeruleus alpha were suppressed by electrical stimulation of the nucleus reticularis pontis oralis in the intensity range 20-100 microA in a stimulus-intensity-dependent manner. These findings indicate that the nucleus reticularis pontis oralis has an inhibitory influence on the functions of the micturition center.

Animals↗
PubMed1995

Regulation of early T cell development by the engagement of TCR-beta complex expressed on fetal thymocytes from TCR-beta-transgenic scid mice.

Transgenic expression of the beta-chain of T cell antigen-receptor (TCR) is known to induce the generation of CD4+ CD8+ thymocytes in the immunodeficient scid mouse, in which thymocyte development is otherwise arrested at CD4- CD8- cells. It is not clear, however, whether or not the thymocyte development is controlled by ligand engagement of the TCR-beta complex on the cell surface. In the present study, we have examined how the engagement by Ab of the TCR-beta complex expressed on the TCR-beta-transgenic scid fetal thymocytes can regulate the generation of CD4+ CD8+ thymocytes. Organ cultures of CD4- CD8- day 14 fetal thymocytes from the TCR-beta-transgenic scid mice resulted in the generation of CD4- CD8+ and then CD4+ CD8+ cells. The initial step from CD40- CD8- cells to CD4- CD8+ cells was enhanced by the addition of anti-TCR-beta Ab, whereas the subsequent step from CD4- CD8+ cells to CD4+ CD8+ cells was markedly inhibited by anti-TCR-beta Ab. These results indicate that ligand engagement of the TCR-beta complex can positively and negatively regulate the early thymocyte development. Moreover, the finding that engagement of TCR-beta complex inhibits the generation of CD4+ CD8+ cells suggests that the induction of CD4+ CD8+ thymocytes by the TCR-beta transgene is not an immediate consequence of cell-surface engagement of the TCR-beta complex but requires liberation from the continued TCR-beta signaling.

Animals↗
PubMed1995

A boundary of long-range G + C% mosaic domains in the human MHC locus: pseudoautosomal boundary-like sequence exists near the boundary.

The human genome is composed of long-range G+C% (GC%) mosaic structures related to chromosome bands. We found the human MHC locus to be an example of megabase-level GC% mosaic structures and predicted a possible boundary of the megabase-level domains within an undercharacterized 450-kb region harboring the junction of MHC classes II and III. Chromosome walking of the 450-kb region and base-compositional analysis precisely located the boundary of the mosaic domains, disclosing a sharp GC% transition. Near the transition point there was a 20-kb dense Alu cluster, a 30-kb dense LINE-1 cluster, and a sequence highly homologous with the pseudoautosomal boundary of the short arms of human sex chromosomes (PAB1X and PAB1Y); PAB1X and PAB1Y are the interface between sex-specific and pseudoautosomal regions. Many PAB1XY-like sequences (PABLs) were detected by hybridization against genomic DNA, and the new sequences defined the complete form of PABLs to be about 650 nt.

Base Composition↗
PubMed1995

Separation and characterization of 25-hydroxyvitamin D3 3-sulfate in human plasma by high-performance liquid chromatography.

The separation and characterization of 25-hydroxyvitamin D3 3-sulfate in human plasma are carried out by high-performance liquid chromatography. The vitamin D sulfate fraction is obtained from a plasma specimen (three volunteers) by the combined use of a Sep-Pak C18 cartridge, for solid-phase extraction, and a lipophilic gel (piperidinohydroxypropyl Sephadex LH-20), for ion-exchange chromatography. 25-Hydroxyvitamin D3 3-sulfate is identified in the examined three specimens in two ways: its chromatographic behavior and that of its fluorescent labeled derivative using 4-[4-(6-methoxy-2-benzoxazolyl)phenyl]-1,2,4-triazoline-3,5- dione; and data obtained from the solvolysis reaction.

Calcifediol↗
PubMed1994

Three genes in the human MHC class III region near the junction with the class II: gene for receptor of advanced glycosylation end products, PBX2 homeobox gene and a notch homolog, human counterpart of mouse mammary tumor gene int-3.

Cosmid walking of about 250 kb from MHC class III gene CYP21 to class II was conducted. The gene for receptor of advanced glycosylation end products of proteins (RAGE, a member of immunoglobulin superfamily molecules), the PBX2 homeobox gene designated HOX12, and the human counterpart of the mouse mammary tumor gene int-3 were found. The contiguous RAGE and HOX12 genes were completely sequenced, and the human int-3 counterpart was partially sequenced and assigned to a Notch homolog. This human Notch homolog, designated NOTCH3, showed both the intracellular portion present in the mouse int-3 sequence and the extracellular portion absent in the int-3. It thus corresponds to the intact form of a Notch-type transmembrane protein. About 20 kb of dense Alu clustering was found just centromeric to the NOTCH3.

Amino Acid Sequence↗
PubMed1994

Effects of MK-801 and CNQX, glutamate receptor antagonists, on bladder activity in neonatal rats.

This study was undertaken to examine the role of N-methyl-D-aspartate (NMDA) and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) glutamatergic receptors in the regulation of urinary bladder activity in the neonatal rat. Experiments were conducted using an in vitro spinal cord-bladder (SB) preparation from 1- to 5-day-old rats or awake neonatal rats 6 and 7 days old. SB preparations were isolated under hypothermic anesthesia. Isovolumetric bladder contractions occurred spontaneously, were induced by electrical stimulation (ES) of the bladder wall or were evoked reflexly by perineal tactile stimulation (PS). MK-801 (3-30 microM), an NMDA receptor antagonist, enhanced the amplitude of spontaneous, ES- and PS-evoked contractions. Removal of the spinal cord after MK-801 abolished PS-evoked reflex contractions but did not change the amplitude of spontaneous and ES-evoked contractions. Removal of the spinal cord in the absence of MK-801 increased the amplitude of spontaneous and ES-evoked contractions, indicating that the bladder is subject to a tonic inhibitory control originating in the spinal cord. 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX, 3-30 microM), an AMPA receptor antagonist, decreased the amplitude of PS-evoked contractions and the frequency of spontaneous contractions in the SB preparation. Removal of the spinal cord after CNQX enhanced the amplitude of spontaneous and ES-evoked contractions but abolished PS-evoked contractions. The frequency of spontaneous contractions which decreased after CNQX increased to near control levels after removal of the spinal cord. In awake neonatal rats, intraperitoneal injection of MK-801 (3 mg/kg) induced spontaneous micturition. A large dose of CNQX (30 mg/kg) decreased PS-evoked micturition volume. These results suggest that NMDA glutamatergic receptors are involved in a lumbosacral spinal inhibitory mechanism controlling bladder activity; whereas AMPA glutamatergic receptors are involved in the perineal-to-bladder reflex pathway in neonatal rats.

6-Cyano-7-nitroquinoxaline-2,3-dione↗
PubMed1994

Electron microscopic and histochemical studies of the mononuclear odontoclast of the human.

BACKGROUND: Osteoclasts and odontoclasts are multinucleated giant cells which resorb hard tissue by the ruffled borders. Recently, the authors reported the presence of a mononuclear osteoclast with a ruffled border in vitro. However, its presence in vivo has not been shown. To demonstrate the presence of a mononuclear odontoclast in humans, the present study used human deciduous teeth. METHODS: After fixation and decalcification, tartrate-resistant acid phosphatase (TRACPase) activity was detected with the azo dye method, and then TRACPase-positive cells were observed on resorbing areas of teeth. TRACPase-positive cells could be distinguished from other cells by light microscopy, and the cells for investigation were serially sectioned by alternating semithin and ultrathin sections to observe their ultrastructure and three-dimensional organization. RESULTS: TRACPase activity was detected in both multinucleated odontoclasts and a mononuclear cell from serial sections. By electron microscopy, most of the multinucleated odontoclasts had ruffled borders and clear zones. A mononuclear TRACPase-positive cell with a ruffled border and clear zone was reconstructed three-dimensionally by NIKON COSMO-ZONE 2SA. The reconstruction showed that this cell had one irregularly shaped nucleus and a wide ring-shaped clear zone and a small ruffled border. Under the ruffled border, this cell formed a small lacuna on the dentin surface. The results suggested that this cell was a mononuclear odontoclast. CONCLUSIONS: The present study concludes that cells with ruffled borders and clear zones observed by transmission electron microscopy can be identified as odontoclasts or osteoclasts irrespective of the number of nuclei.

Acid Phosphatase↗
PubMed1994

Cat and dog: higher center of micturition.

The nervous control of micturition in the pontine level was studied using cats and dogs as research specimens. Two techniques for this purpose were employed: (1) electrical and chemical stimulations research and (2) nerve tracing research. This investigation suggests that two distinct pontine centers exist, the commonly known pontine micturition center (PMC) which is responsible for urine emptying, and a second, very important pontine center which functions as the urine storage facilitator (PUSFC). The PMC, which triggers bladder contractions and micturition, is located in the nucleus locus coerleus alpha (LCa). The second center (PUSFC) is located in the nucleus locus subcoerleus (LSC). Micturition is controlled by close coordinate operation of both centers in the pontine level.

Animals↗
PubMed1994

Recombination in the class III region of the mouse major histocompatibility complex.

The sites of meiotic recombination in the class II region of the mouse major histocompatibility complex (MHC) are clustered at hotspots. To search for hotspots in the class III region, we mapped recombinational breakpoints of 79 Ab:H2-D recombinants with 11 DNA markers; these included Tnx, the gene for an extracellular matrix protein, tenascin X, the Notch-related Int3 gene, and a microsatellite marker, D17Mit13, none of which had previously been mapped precisely. The results gave the gene order of Eb-61.1-Int3-Tnx-Cyp21/C4-Bf-Hsp68c-D17Mit13+ ++-Tnfa/Tnfb-D. The crossover sites in 40 of the 79 recombinants were confined within the Eb/Int3:Tnx/Cyp21 interval. The result demonstrated that an unequal distribution of recombination is a general feature of the mouse MHC, suggesting the presence of a recombinational hotspot within the Int3:Tnx interval.

Animals↗
PubMed1994

Automated collection of conditioned medium from organotypically cultured brain slice.

For the analysis of neural activity over a long period of time, the organotypic culture of mammalian brain slices provides excellent specimens. To effectively utilize the slice culture, we developed a device for automatic sampling of the culture medium. This device is a computer-controlled combination of a multichannel peristaltic pump to remove the media from the glass culture containers, a fraction collector designed to allow quick freezing of the samples, and a multichannel syringe pump to deliver new media to the containers. Using this device, substances released as results of neural activities can be collected at regular intervals over several days. We monitored the circadian release of arginine-vasopressin from cultured suprachiasmatic nuclei. We also monitored tonic releases of lactate dehydrogenase from cultured hippocampi phasically treated with an excitotoxin and from those transiently deprived of oxygen/glucose.

Animals↗
PubMed1994

Nitric oxide synthase gene expression in cholinergic neurons in the rat brain examined by combined immunocytochemistry and in situ hybridization histochemistry.

The expression of mRNA for the calmodulin-dependent form of brain nitric oxide synthase (NOS) was examined in cholinergic cells of the rat brain using a method combining in situ hybridization histochemistry with immunocytochemistry for choline acetyltransferase (ChAT) in the same brain sections. We constructed a riboprobe specific for brain NOS by subcloning a 493 bp fragment of the coding region which displayed low homology to other forms of NOS. The general distribution of NOS mRNA was in excellent agreement with previous studies using the full-length probe or NADPH diaphorase histochemistry. NOS mRNA was observed in many brain structures and relative levels were quantitated using grain counting procedures in a number of cholinergic and non-cholinergic neuronal groups throughout the brain. In the forebrain, ChAT-immunoreactive cells or cell groups were observed in medial septum (MS), vertical limbs of diagonal band (DBV) and horizontal limbs of diagonal band (DBH), nucleus basalis magnocellularis (NBM), substantia innominata (SI), and striatum (ST). In the brainstem, the cholinergic groups studied included those located in the pedunculopontine tegmental nucleus (PPTN), the laterodorsal tegmental nucleus (LDTN), the nucleus parabigeminalis and several motor nuclei. For NOS mRNA quantitation, silver grains overlying ChAT-stained neuronal profiles in sections on emulsion-dipped slides were counted digitally. In the LDTN and PPTN, virtually all the ChAT-positive cells expressed NOS mRNA at high levels. In MS, DBV and SI, about 30-50% of the ChAT-positive cells expressed NOS mRNA at low-to-moderate levels. Less than 20% of ChAT-positive neurons in the other cholinergic populations studied expressed NOS mRNA; the NBM was one of these low-expressing populations. Many scattered non-cholinergic cells expressing NOS mRNA were found in the striatum and cerebral cortex. In other non-cholinergic regions, high NOS mRNA expression was observed in the islands of Calleja, thalamic and hypothalamic nuclei, several amygdaloid nuclei, regions related to the optic tract, the interpeduncular nucleus, and the supramammillary nucleus. The heterogeneous distribution of NOS mRNA implies complex roles for nitric oxide neurotransmission in brain function, including for the cholinergic phenotype. Additionally, given the postulated involvement of nitric oxide in neurodegeneration, the widely varying levels of expression of NOS within identified central cholinergic neurons may relate to differential vulnerability of this phenotype in disease or aging.

Amino Acid Oxidoreductases↗
PubMed1994

Micturition reflexes in the in vitro neonatal rat brain stem-spinal cord-bladder preparation.

An in vitro neonatal (1-7 day) rat brain stem-spinal cord-bladder (BSB) preparation was used to examine the central control of micturition. Isovolumetric bladder contractions occurred spontaneously or were induced by electrical stimulation of the ventrolateral brain stem, spinal cord, bladder wall (ES-BW), or by perineal tactile stimulation (PS). Transection of the spinal cord at the L1 segment increased the amplitude of ES-BW- and PS-evoked contractions, and subsequent removal of the spinal cord further increased spontaneous and ES-BW-evoked contractions but abolished PS-evoked contractions. Hexamethonium (1 mM), a ganglionic blocking agent, mimicked the effect of cord extirpation. Tetrodotoxin (1 microM) blocked ES-BW- and PS-evoked contractions but enhanced spontaneous contractions. Bicuculline methiodide (10-50 microM), a gamma-aminobutyric acid A receptor antagonist, increased the amplitude of spontaneous, ES-BW- and PS-evoked contractions. These results indicate that PS-evoked contractions are mediated by spinal reflex pathways, whereas spontaneous and ES-BW-evoked contractions that are elicited by peripheral mechanisms are subject to a tonic inhibition dependent on an efferent outflow from the spinal cord. PS-evoked micturition is also subject to inhibitory modulation arising from sites rostral to the lumbosacral spinal cord. Although electrical stimulation of bulbospinal excitatory pathways can initiate bladder contractions in the neonatal rat, these pathways do not appear to have an important role in controlling micturition during the first postnatal week.

Animals↗
PubMed1994

Tissue kallikrein in rat and mouse neurons.

1. The distribution of tissue kallikrein in the rat brain was investigated by an immunohistochemical technique using antiserum against rat urinary kallikrein. The kallikrein-immunoreactive cells were widespread and scattered in both the cerebral cortex and brain stem, and the immunoreactive substance appeared to be preferentially localized around the neuronal cell body and their processes. 2. Furthermore, in order to define whether brain kallikrein levels vary in the aging process, the hydrolyzing activity in the cerebrum L-prolyl-L-phenylalanyl-L-arginine-4-methyl-coumaryl-7-amide (Pro-Phe-Arg-MCA) was measured in young (7-week old) and old (36-week old) female senescence accelerated mice (SAM-P/8 and SAM-R/1). The brain kallikrein levels of 7- as well as 36-week old SAM-P/8 were found to be lower than those of 7-week old SAM-R/1 (a control strain). Also, the brain kallikrein level of 36-week old SAM-R/1 animals was markedly reduced when compared with the 7-week old mice, suggesting that the reduced amount of cerebral kallikrein reflects the aging of the brain.

Aging↗