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Biomedical subjects

K Watabe

Publications and source records attributed to K Watabe.

At least 127 records · Page 7Linked to original sources
PubMed1986

Immunochemical similarity of synthetic alpha-(1----3)-branched D-glucans to dextran B1375.

Anti-dextran B1375 antibodies were raised in rabbits by injecting formalin-killed Leuconostoc mesenteroides strain NRRL B1375, and the anti-dextran serum was used to examine native dextran B1375, and synthetic linear and four alpha-(1----3)-branched alpha-(1----6)-D-glucopyranans for similarities. The antiserum reacted with the homologous dextran B1375 and also with all the synthetic linear and branched glucans. Precipitation and precipitation-inhibition studies indicated that the antiserum contained at least three groups of antibodies with different specificities, the first specific for linear alpha-(1----6)-D-glucopyranan structure, the second specific for alpha-D-glycopyranosyl-(1----3)-branching and the last specific for another, unknown structure present in the dextran B1375 molecule. Two samples of the synthetic branched glucans were shown to be immunochemically the most similar to natural dextran B1375 by inhibition experiments.

Antibodies↗
PubMed1985

The characteristics of the antidromic discharge of cat dorsal raphe neurons during repetitive activation.

Slowly discharging neurons in the cat dorsal raphe could be classified into 3 types according to the behavior of antidromic spike discharges during repetitive stimulation of the medial forebrain bundle at 10 Hz. In the types 1 and 2, the latency of antidromic discharge was gradually prolonged to reach an asymptote, whereas no marked change occurred in the type 3. The type 2 neurons, which had a slower conduction velocity, showed a greater prolongation than the type 1 neurons. The maximum length of this prolongation was not significantly correlated with the initial latency. During 10 Hz stimulation some neurons showed repeatedly a conduction block after a sequence of initial decrease and later increase in latency. The spontaneous discharge was strongly suppressed during 10 Hz stimulation. During 1 Hz stimulation just after the cessation of 10 Hz stimulation, the prolonged antidromic latency was gradually restored in parallel with the recovery of the spontaneous discharge. Circumstantial evidences seem to be in favor of the idea that hyperpolarization of the axonal and somatic membranes is mainly responsible for the observed behavior of antidromic spikes of type 1 and 2 neurons.

Animals↗
PubMed1985

Developmental microvascular architecture of the rat cerebellar cortex.

External and internal microvascular architectures of the developing rat cerebellar cortex, from embryonic day 18 to postnatal day 14 and in adults, were studied using a cerebrovascular casting method for scanning electron-microscopic observation. The external vascularization of the developing cerebellum showed the most significant alteration in vascular morphology at the stage of intensive proliferation of matrix cells in the external granular layer (EGL) from birth to postnatal day 4. It consisted of multiple luminal protrusion of the vessels, septum formation in the lumina, and small, ring-like anastomoses. Moreover, at the end of this stage, these structures of the vessels disappeared and the subarachnoid space was filled with newly-formed microvascular networks. Thereafter, architectural change of the developing pial vessels was mainly accomplished by elongation of each contorted vessel of the network. Concerning internal vascularization, a few vessels connected with the pial vessels were observed in the cerebellar plate forming a loose, simple network in the deeper neural parenchyma before the stage of foliation began. During the period of thickening of the EGL, however, there was no alteration in vascularity of the parenchyma other than the architectural changes proportionate to the newly-formed folia. It was during the synaptogenetic stage in the internal granular layer that the earliest intraneural vascular plexuses were formed. The vascular network in the molecular layer was formed after disappearance of the EGL. These findings suggest that vascular proliferation correlated with EGL-formation pertains to the pial vessels, and not the intraneural ones, which develop after neuronal cell migration in the developing cerebellum has taken place.

Animals↗
PubMed1985

An immunohistochemical study of aldolase C in normal and neoplastic nervous tissues.

Cellular localization of aldolase C isozyme was examined in the normal human brain and in nervous tissue tumors by the indirect immunoperoxidase method using an antibody to aldolase C. In normal brain tissues, staining was most prominent in astrocytes and Purkinje cells, although faint staining was also occasionally observed in some other neurons. Oligodendroglia and ependymal cells showed no distinct staining. The nervous tissue tumors we examined included 34 gliomas (ten astrocytomas and 24 anaplastic gliomas), 30 medulloblastomas, and seven neuronal tumors. Positive staining was observed in some gliomas, but not in the medulloblastomas and neuronal tumors examined. In gliomas, nine of ten astrocytomas and six of 24 anaplastic gliomas showed positive staining in their main constituent cells. In the remaining gliomas, most constituent cells were unstained or positive cells were only sporadically present. These findings indicated that aldolase C was expressed in some of astrocytic glioma cells as in the normal counterpart, more frequently in more differentiated form.

Brain↗
PubMed1985

Glial fibrillary acidic protein in medulloblastomas.

Two cases of medulloblastoma are described which show positive immunostaining for glial fibrillary acidic protein (GFAP) in many cells. The surgical and autopsy specimens were examined by the indirect immunoperoxidase method. Positive staining for GFAP was demonstrated in the small, round to polygonal cells in both surgical specimens and in the small spindle cells in the autopsy specimen of one case. In the small, round to polygonal cells positive GFAP was shown as a perinuclear brown rim or intracytoplasmic brown droplet. In the small spindle cells, the cytoplasm and the polar processes were stained. Except for GFAP staining, these positive cells were morphologically indistinguishable from the accompanying unstained cells, indicating that GFAP was expressed in the medulloblastoma cells. Considering that GFAP is specific for astrocytes, these findings suggest the potential of astrocytic differentiation in the neoplastic cells of these medulloblastomas. The findings obtained in other 28 medulloblastomas examined in parallel are also discussed briefly.

Biopsy↗
PubMed1985

Two types of rostroventral medulla neurons projecting to the trigeminal spinal nucleus as differentiated by the response to antidromic activation and the histological location.

A population of neurons in the rostroventral medulla, which send their axons to the subnucleus oralis of the trigeminal spinal nucleus of rats, could be differentiated into two types on the basis of their location and the variability of antidromic latency during repetitive stimulation at 10 Hz. Type A neurons were mostly located in the raphe magnus and were activated antidromically with a relatively long latency, which gradually increased during repetitive stimulation. By contrast, type B neurons were located in the medial portion of the gigantocellular reticular nucleus, and responded with a relatively short, stable antidromic latency.

Animals↗
PubMed1985

Plasticity in cat visual cortex restored by electrical stimulation of the locus coeruleus.

It has been proposed that the presence of noradrenaline (NA)-containing terminals and NA-related receptors within the visual cortex is necessary to maintain the high level of neuronal plasticity in the immature visual cortex of kittens. In the present study we wanted to show whether electrical stimulation of the locus coeruleus (LC), which contains the somata of these cortical NA fibers, can restore neuronal plasticity to the normally aplastic visual cortex of juvenile and adult cats. We consistently found a significant loss of binocular cells in the visual cortex of mature animals which had monocular vision for only 12 h dispersed over 6 days (2 h a day, otherwise kept in the dark) in combination with concurrent LC stimulation. This result was interpreted as indicating that endogenous NA released from NA terminals restored susceptibility to monocular vision in the mature visual cortex. We next examined how long the restored plasticity lasts in the same animals after the LC stimulation was ended. The animals revived from the first recording session were either returned to the same daily schedule of brief monocular exposure (light/dark = 2/22 h) as before, or subjected to the usual monocular lid suture and kept in a cat colony environment (light/dark = 16/8 h). The LC electrodes had been removed and no more electrical stimulation was delivered at this stage. In the animals subjected to reiteration of brief monocular exposure, the state of reduced binocularity gradually returned to normal over a period of 2-3 weeks after stopping LC stimulation. We calculated that the revived plasticity disappeared at an average rate of a 22% loss every 7 days. This result sharply contrasted with the result obtained in the animals subjected to usual monocular lid suture. In this test the state of reduced binocularity continued for at least the next 3 weeks, suggesting that the restored plasticity was sustained throughout a period of 3 weeks (longest term tested). The different results obtained in the two paradigms may be explained by the different strength of binocular imbalance in the two tests imposed on the visual cortex in which neuronal plasticity was restored partially.

Animals↗
PubMed1985

Antidromic discharge property of meso-accumbens dopaminergic VTA neurons in rats.

Three groups of meso-accumbens (Acc) neurons in the ventral tegmental area were differentiated by their antidromic discharge property; dopaminergic type 1 (n = 10), non-dopaminergic type 2 (n = 2) and unclassified (n = 2) neurons. During repetitive activation at 10 Hz, the latency of the initial segment (IS) spike, which was often not followed by the somadendritic (SD) spike, was gradually prolonged in type 1, but not in type 2 and unclassified neurons. The latency prolongation of type 1 neurons was reduced to about a half of the normal in rats treated with kainic acid plus haloperidol, but only slightly when treated with kainic acid or picrotoxin. The rate of SD invasion tended to increase after all kinds of chemical treatment. Stimulation of the medial forebrain bundle in type 1 neurons gave responses comparable to Acc stimulation. It is suggested that the latency prolongation of IS spike is produced mainly by axonal mechanism. But additional somatic mechanisms such as dopaminergic self-inhibition and GABAergic and non-GABAergic inputs from the Acc would make some contribution, and at the same time produce frequent suppression of the antidromic SD spike.

Action Potentials↗
PubMed1985

Immunohistochemical study on neuroglia identified by the monoclonal antibody against a macrophage differentiation antigen (Mac-1).

We have studied frozen sections of the developing and adult mouse central nervous system (CNS), with or without cold lesions, by immunohistochemical and histochemical methods. Using a monoclonal antibody against a macrophage differentiation antigen (Mac-1), we have shown that some neuroglia in the white matter of adult mice stained positively. In the developing CNS, with or without cold lesioning, Mac-1-positive glia were not detected. In the normal adult CNS, a small number of glia in the white matter stained faintly. After cold injury, the number of Mac-1-positive glia and their staining intensity increased for several months. Mac-1-positive glia were always negative for glial fibrillary acidic protein (GFA). Their morphology and distribution were similar to those of nucleoside diphosphatase-positive cells. Considering that the phagocytic activity of glia increases after injury to the CNS (Trachtenberg 1983) and that Mac-1 has been reported to be associated with the complement receptor (Beller et al. 1982), Mac-1-positive glia may play a role in phagocytosis in the damaged CNS.

Animals↗
PubMed1985

[Immunohistochemical and ultrastructural studies on tactile-like corpuscles in neurofibromas].

Morphological characteristics of tactile-like corpuscles (tactoid bodies, pseudomeissnerian corpuscles) which are occasionally present in neurofibromas have already been detailed. However, there has yet been controversy on the cytogenesis of these lamellated structures. Probably, one of the most important problems is whether tactile-like corpuscles are composed of Schwann cells or of perineurial cells. In this study, seven cases of neurofibromas with tactile-like corpuscles were examined by rabbit or mouse polyclonal antisera to S-100 protein which could be regarded, at least in the peripheral nervous system, as a specific marker for Schwann cells. Since recent extensive studies have revealed that S-100 protein is a mixture of two predominant dimeric components with S-100 alpha and S-100 beta subunits, and the S-100 alpha subunit is absent from Schwann cells, tactile-like corpuscles were also examined by a mouse monoclonal antibody (ASA-1) specific for the S-100 alpha subunit. For immunohistochemical analysis, peripheral nerves, Meissner corpuscles and epidermal melanocytes were also examined in parallel. By the immunoperoxidase method using rabbit or mouse polyclonal antisera to S-100 protein, tactile-like corpuscles were intensely stained in sections from all seven cases. Both the flattened cell bodies and the eccentrically located nuclei of their constituent cells were stained. In the background neurofibroma tissue surrounding the tactile-like corpuscles, spindle-shaped cells were stained variably. In three of the seven cases, numerous melanin-containing cells which often surrounded the tactile-like corpuscles were also stained. In normal peripheral nerves, positive staining was confined to Schwann cells and their processes.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1984

A 3' to 5' exonuclease activity is associated with phage 029 DNA polymerase.

Bacteriophage 029 produces its own DNA polymerase which is encoded by gene 2 [Watabe, K. and Ito, J. (1983) Nucleic Acid Res. 11, 8333]. This 029 DNA polymerase has been purified by phospho-cellulose, DEAE-cellulose, double-stranded DNA cellulose chromatography and glycerol gradient centrifugation. An exonuclease activity associated with the DNA polymerase was found through all the steps of the purification. This nuclease preferably degrades single-stranded DNA from the 3' to the 5' terminus direction, suggesting that the enzyme plays a role for proofreading during DNA replication. While DNA polymerase activity isolated from cells infected with temperature sensitive mutant of gene 2 is thermolabile, the nuclease activity is not significantly reduced at the restrictive temperature.

Bacillus subtilis↗
PubMed1984

Antibodies specific for the phi 29 terminal protein inhibit the initiation of DNA replication in vitro.

The phi 29 DNA-terminal protein serves as a primer for the initiation of DNA replication by covalently binding the first nucleotide in the DNA chain. Two distinct antibodies were used for functional analysis of this protein. One antibody was raised against sonicated phi 29 DNA-protein complex isolated from phage virions (anti-TP). The other antibody was raised against a conjugate of bovine serum albumin and a synthetic peptide corresponding to the carboxy-terminal of the phi 29 terminal protein (anti-gp3C), which was predicted from the nucleotide sequence of phi 29 DNA. Both antibodies react with native phi 29 terminal protein as determined by immunoprecipitation and enzyme-linked immunosorbent assay. Both antibodies specifically inhibit the complex-forming reaction between the phi 29 terminal protein and dAMP, the first nucleotide of phi 29 DNA.

Amino Acid Sequence↗
PubMed1984

Replication of bacteriophage phi 29 DNA in vitro: the roles of terminal protein and DNA polymerase.

phi 29 DNA replication is initiated by the formation of a covalent complex between the viral-coded terminal protein and dAMP (TP-dAMP). This initiation reaction system has been reconstituted from two phage-encoded proteins, the terminal protein and DNA polymerase. The phi 29 DNA polymerase was purified from phage-infected cells by using poly(dA) X p(dT)12-18 as an assay template. The purified polymerase has an apparent molecular mass of 68 kDa in its native form and it appears to function as a monomer. The terminal protein was purified to homogeneity from Escherichia coli cells harboring a cloned plasmid that contained a phi 29 gene 3 segment. The molecular mass of the purified terminal protein was about 30 kDa in both the denatured and the native form. The protein apparently functions as a monomer. When the terminal protein and DNA polymerase were incubated in the presence of dATP, Mg2+, and phi 29 DNA-protein as template, the terminal protein bound covalently to dAMP. This reaction did not require ATP. In addition, these two purified fractions catalyzed DNA chain elongation from both ends of phi 29 DNA, yielding the expected 9- to 12-base fragment when assayed in the presence of 2',3'-dideoxycytidine triphosphate. These results indicate that phi 29 DNA polymerase catalyzes formation of the terminal protein-dAMP complex and can also catalyze chain elongation at least 9-12 bases from both ends of phi 29 DNA.

Bacillus subtilis↗
PubMed1984

Increase of translatable mRNA for major microsomal proteins in n-alkane-grown Candida maltosa.

In an n-alkane-assimilating Candida sp., transfer from glucose- to n-alkane-containing medium induced changes in the microsomal proteins, and several distinctive polypeptides were demonstrated in the solubilized microsomal fraction derived from n-alkane-grown cells. Long-term-labeling and pulse-labeling experiments in vivo demonstrated the synthesis of the specific microsomal polypeptides. The polypeptides were synthesized as in vitro translation products directed by polyadenylated RNA extracted from n-alkane-grown cells. Two major polypeptides were partially purified from the microsomal fraction from n-alkane-grown cells, and antiserum was prepared in a rabbit. Immunoprecipitation of these two polypeptides was accompanied by an increase in the amount of translatable mRNA. The molecular weights of the polypeptides derived from long-term-labeling, pulse-labeling and in vitro translation experiments appeared to be identical.

Alkanes↗
PubMed1983

A novel DNA polymerase induced by Bacillus subtilis phage phi 29.

A novel DNA polymerase induced by Bacillus subtilis bacteriophage phi 29 has been identified. This polymerase can be separated from host DNA polymerase, by fractionation of extracts prepared from phage infected cells, using phosphocellulose chromatography. The isolated polymerase prefers poly(dA)oligo(dT) as template. The DNA polymerase isolated from the cells infected with a gene 2 temperature sensitive mutant (ts2) showed greater heat-lability than that induced by wild type phi 29. The ts2 DNA polymerase was also thermolabile for its activity in the formation of a covalent complex between phi 29 terminal protein and dAMP, the initiation step of phi 29 DNA replication. These findings indicate that gene 2 is the structural gene for a phi 29 DNA polymerase required for the complex formation step of DNA initiation.

Bacillus subtilis↗
PubMed1983

Some dorsal raphe axons of the cat bifurcate to project into bilateral medial forebrain bundles.

Seventy-one dorsal raphe (DR) neurons of the cat were antidromically activated by electrical stimulation of the medial forebrain bundle (MFB). It was found that not a small number of DR neurons send their axons contralaterally. Seven out of 67 DR neurons (10.4%) responded to stimulation of both sides of MFB. Positive collision test for the bilaterally evoked responses indicates that single axons of these DR neurons bifurcate to innervate bilateral forebrain structures.

Animals↗
PubMed1983

Revised pathophysiology on BBB damage: the edema as an ingeniously provided condition for cell motility and lesion repair.

Probably, the most important evidence concerning the breakdown of the BBB is a large inflow of hematogenous fluid into the extravascular spaces. Thus all parenchymal cell elements represent freely floating cells in this fluid medium called the edema fluid. These essential morphological alterations, such as extremely expanded extracellular space and freely floating cells within the fluid, were also observed in the developing normal fetal brain. Many neuroblasts were vigorously migrating in the fetal brain, as were the macrophages and reactive astrocytes in the edema fluid. Obviously, hematogenous cells and reactive astrocytes in the edematous lesion take part in its repair. Many astrocytes, GFAP positive, in the 3 or 4 day-old lesions, revealed mitosis. And in vitro, when we immersed these astrocytes in the protein and glucose rich medium, they demonstrated a remarkably changed morphology and were moved into the M and G1 phases, thus gaining the ability of cell motility. This was also true in the edema fluid. Brain edema is definitely a serious "pathological" condition. But it is also conceivable from a different biological aspect that as a result of BBB "opening", free extracellular space essential for cell motility and a source of their energy is ingeniously provided for these cells, and thus the lesion can be effectively repaired. The biological significance of the edema fluid was emphasized and stereotaxic morphology and cinematography, supporting the above evidence, have also been presented.

Animals↗