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S Kashiwamata

Publications and source records attributed to S Kashiwamata.

At least 37 records · Page 2Linked to original sources
PubMed1989

When does GABA-like immunoreactivity appear in the rat cerebellar GABAergic neurons?

The time of appearance of gamma-aminobutyric acid (GABA), a well-known neurotransmitter, during the development of cerebellar GABAergic neurons in rats was investigated immunocytochemically using purified anti-GABA antibody. Sprague-Dawley rats were used at embryonic days 15, 16, 18, 19 and 21, and postnatal days 0, 1, 5, 10, 15, 20 and 30. Golgi cells showed processes and GABA-like immunoreactivity at embryonic day 16 during migration. Purkinje cells were found immunoreactive at embryonic day 18, when they arrived at their destination. The reactivity of the basket cell was already apparent at postnatal day 5, and was thought to appear just after the end of migration. In all of the GABAergic neurons, GABA-like immunoreactivity was visible much earlier than the time of synapse formation and the emergence of their electrophysiological activity described in the literature. In addition, GABA-like immunoreactivity tended to shift from the soma and dendrite into the axon with development.

Aging↗
PubMed1989

Critical period of bilirubin-induced cerebellar hypoplasia in a new Sprague-Dawley strain of jaundiced Gunn rats.

Homozygous (j/j) and heterozygous (j/+) newborn Gunn rats of the Sprague-Dawley strain were photoirradiated for 24 h at scheduled postnatal days and the effects of irradiation on the cerebellar development were examined at 30 days of life. Improvement of the survival rate was the most notable effect of photoirradiation. A single 24-h dose of photoirradiation during a period of postnatal days 4-11 effectively prevented hypoplasia in the j/j rat cerebellum. No prevention by light was observed at days 3 and 12. It was found that the most effective day of irradiation on the cerebellar development of j/j rats was centered on postnatal day 7. When plasma bilirubin was assessed during the period of postnatal days 7-10, a distinct diminution of the concentration was observed, restricted to only the period of the light treatment. Although there were some differences in the effective day as well as in the degree of efficacy of phototherapy among cerebellar lobules or sublobules, day 7 was the most critical for cerebellar hypoplasia due to bilirubin.

Animals↗
PubMed1989

Glycosaminoglycan composition of PC12 pheochromocytoma cells: a comparison with PC12D cells, a new subline of PC12 cells.

PC12D cells, a new subline of conventional PC12 cells, respond not only to nerve growth factor but also to cyclic AMP by extending their neurites. These cells are flat in shape and are similar in appearance to PC12 cells that have been treated with nerve growth factor for a few days. In both cell lines, we have characterized the glycosaminoglycans, the polysaccharide moieties of proteoglycans, which are believed to play an important role in cell adhesion and in cell morphology. Under the present culture conditions, only chondroitin sulfate was detected in the media from PC12 and PC12D cells, whereas both chondroitin sulfate and heparan sulfate were found in the cell layers. The levels of cell-associated heparan sulfate and chondroitin sulfate were about twofold and fourfold higher in PC12D cells than in PC12 cells, respectively. Compared to PC12 cells, the amounts of [35S]sulfate incorporated for 48 h into chondroitin sulfate were twofold lower but those into heparan sulfate were 35% higher in PC12D cells. The amount of chondroitin sulfate released by PC12D cells into the medium was about a half of that released by PC12 cells. The ratio of [35S]sulfate-labeled heparan sulfate to chondroitin sulfate was 6.2 in PC12D cells and 2.2 in PC12 cells. These results suggest that there may be some correlation between the increase in content of glycosaminoglycans and the change in cell morphology, which is followed by neurite outgrowth.

Adrenal Gland Neoplasms↗
PubMed1989

Sex-dependent and sex-independent distribution of the beta-subunit of nerve growth factor in the central nervous and peripheral tissues of mice.

Levels of the beta-subunit of nerve growth factor (beta-NGF) were measured in the central nervous and peripheral tissues of mice using a highly sensitive, sandwich-type enzyme immunoassay system. Antiserum was raised in rabbits against the 7S form of NGF, which was purified from mouse submandibular glands. beta-NGF-specific antibody isolated on a column of Sepharose CL-4B coupled with purified beta-NGF reacted only with beta-NGF. The assay for beta-NGF was performed by incubation of F(ab')2 fragments of the antibody immobilized on a polystyrene ball with tissue extract and then with the same antibody Fab' fragments labeled with beta-D-galactosidase, followed by measurement of galactosidase activity. Our assay system was found to be highly sensitive (minimal detection limit, 0.3 pg/0.3 ml of assay mixture). Furthermore, the presence of gelatin hydrolysates and protease inhibitors during preparation of tissue extracts enabled us to determine the precise levels of beta-NGF in almost all organs of mice. The amount of beta-NGF in submandibular glands was extremely high, and its level increased rapidly until mice were 2 months of age; then, the level continued to increase slowly until mice were 1 year old (3-5 mg/g of tissue). In serum, some of the 2-month-old males, but none of the females, exhibited a fairly high level of beta-NGF (greater than 100 pg/ml).(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Glands↗
PubMed1989

Mode of bilirubin deposition in the cerebellum of developing jaundiced Gunn rats.

Bucolome is known as a potent displacer of bilirubin from the blood into tissues. The effects of the drug on newborn rats with congenital jaundice (Gunn rats) were examined on postnatal days 7, 11, 15 and 21. A single subcutaneous injection of bucolome resulted in a rapid fall of the total plasma bilirubin concentration and the lowered level persisted as long as 23 h. Concomitantly with the drop, the cerebellar bilirubin level increased within 1 h. Although cerebellar bilirubin returned to the initial level after 24 h in rats treated with bucolome on days 7 and 21, it remained almost constant or rather increased during the period of 1-24 h in rats treated on days 11 or 15. In 15-day-old rats, localized yellow staining in the cerebellum 24 h after treatment was most apparent in the granule cell layer of the ventral part of the pyramis and the dorsal part of the uvula under the dissecting microscope. These observations suggest that the period most susceptible to bilirubin deposition lies around day 15 in the cerebellum of jaundiced Gunn rats.

Aging↗
PubMed1988

Glycine-like immunoreactivity in the rat auditory pathway.

From neurophysiological and biochemical studies it has been suggested that glycine can function as a major inhibitory neurotransmitter in the central nervous system of mammals. In the present study, anti-glycine antiserum was obtained from rabbits immunized with glycine conjugated to rabbit serum albumin via glutaraldehyde and purified by affinity chromatography. The antibody thus obtained was found specific for glycine as determined by an enzyme immunoassay system. The immunocytochemical distribution of glycine in the auditory tract and internal ear was investigated with the antibody. In the central auditory pathway, glycine-like immunoreactivity was mainly located in the ventral and dorsal cochlear nuclei, trapezoid body, lateral lemniscus and inferior colliculus. In the labyrinth, immunoreactivity was detected in the vestibular ganglion and the supporting cells of the crista ampullaris and the organ of Corti, but not in the spiral ganglion. These findings suggest an important role of glycine in the auditory and vestibular pathways.

Animals↗
PubMed1988

A simple method for preparation of molecular weight marker proteins for sodium dodecyl sulfate-polyacrylamide gel electrophoresis by photopolymerizations of hemoglobin subunits.

Photoirradiation of globin, obtained from human hemoglobin, in the presence of oxygen and protoporphyrin produced a series of polymers of globin subunits ranging from dimer to dodecamer. These polymers are useful as molecular weight markers because they cover a relatively wide range of molecular weights (Mr 15,500-186,000) with a constant and narrow interval of Mr 15,500.

Biomarkers↗
PubMed1988

A possible contribution by glial cells to neuronal energy production: enzyme-histochemical studies in the developing rat cerebellum.

Recent reports have revealed that certain neurons do not survive in vitro in the presence of glucose, which is the primary substrate and exclusive source of energy in the brain. But these neurons can survive in the presence of low-molecular-weight agents such as pyruvate, which are supplied by glial cells (Selak et al. 1984). To test whether this result also holds true in vivo, we investigated the distribution of hexokinase, lipoic dehydrogenase, beta-hydroxybutyrate dehydrogenase, and glucose-6-phosphate dehydrogenase activities in the developing rat cerebellum. Hexokinase activity was relatively higher in glial cells than in neurons. After postnatal day 8, the activity of hexokinase could hardly be detected in Purkinje cells, whereas it was highest in Bergmann glial cells. Purkinje cells were the only type of neuron with high levels of lipoic dehydrogenase at all ages tested. beta-Hydroxybutylate dehydrogenase activity was also high in Purkinje cells, especially in those from young rats. Relatively high glucose-6-phosphate dehydrogenase activity was demonstrated in basket and stellate cells from adult brain. Thus, it appears that, in vivo, certain neurons utilize relatively little glucose, and it is indeed possible that glial cells may supply some substance(s) other than glucose, for example pyruvate, as the primary source of energy.

Aging↗
PubMed1988

Cyclic nucleotides and the activity of glia maturation factor in the hypoplastic cerebellum of developing jaundiced Gunn rats.

Developmental changes of cyclic nucleotides were studied in the hypoplastic cerebellum of jaundiced Gunn rats over the period of postnatal days 8 to 30. The mitogenic activity of glia maturation factor was also measured at day 15. In jaundiced homozygotes (jj), the amount of cyclic GMP on a protein basis was not significantly different from that in control heterozygotes (j+) at either day 8 or 15, but at day 30 it was reduced to about 19% of the control. On the other hand, a lowered nucleotide level on a wet weight basis in jj rats was already statistically significant at day 15. In contrast to cyclic GMP, the rates of increase of cyclic AMP on a wet weight basis were almost the same in the two groups of rats, but the nucleotide levels on a protein basis at days 15 and 30 were a little, but significantly, higher in jj rats than in j+ rats. The activity of glia maturation factor in jj rats was found to be 1.5-3 times as high as that in j+ rats. Possible implications of the present results are discussed.

Animals↗
PubMed1988

Comparative study of glial marker proteins in the hypoplastic cerebellum of jaundiced Gunn rats.

The behavior of marker proteins of glial cells [alpha-enolase, beta-S100 protein, and glial fibrillary acidic protein (GFAP)] was investigated quantitatively by using enzyme immunoassay systems during the development of cerebellar hypoplasia in jaundiced Gunn rats. A neuronal marker protein, gamma-enolase, was also measured as a reference. At postnatal day 8 corresponding to the early stage of cerebellar damage, the amount of beta-S100 on a protein basis was significantly higher in jaundiced homozygotes (jj) than in control nonjaundiced heterozygotes (j+), whereas no differences in alpha- and gamma-enolases and GFAP were observed between the two groups of rats. At days 15 and 30, which correspond, respectively, to the advanced and late stages of cerebellar damage, the three glial proteins, especially GFAP, were higher and the neuronal protein was lower in the jj rat cerebellum than in the control. These results are consistent with the reported histological observations that neuronal cells are vulnerable and damaged by bilirubin, whereas glial cells seem to be less sensitive. On the other hand, the amounts of beta-S100 and alpha-enolase per cerebellum were significantly lower in jj rats at days 15 and 30, as in the case of gamma-enolase, whereas that of GFAP remained at the same level as the control at day 15 and showed a slight but significant decrease at day 30. The possibility is suggested that beta-S100 and GFAP may be available as biochemical indicators of glial cells, especially in the early and advanced stages of cerebellar damage, respectively, but that alpha-enolase is less available.

Animals↗
PubMed1988

Pharmacological and biological effects of tin-protoporphyrin on neonatal hyperbilirubinemic Gunn rats.

Our study was undertaken to examine the pharmacological and biological effects of tin-protoporphyrin, a competitive inhibitor of heme oxygenase, on 5- or 6-day-old homozygous (j/j) Gunn rats with hereditary unconjugated hyperbilirubinemia. When j/j neonates were injected subcutaneously with 20 mumol of tin-protoporphyrin/kg of body weight, hepatic heme oxygenase activity decreased to 30% of the initial level 2 h after administration and remained low during the next 46 h. However, the reduction of serum bilirubin was more rapid and transient, reaching the minimum value (40% of the initial level) at 1 h and increasing thereafter at a rate almost comparable to that in nontreated j/j rats. The mortality rate of j/j rats was strikingly reduced by the administration of 1 to 100 mumol of tin-protoporphyrin/kg; the most effective dose was 5 mumol/kg (8% compared with 80% in non-treated j/j rats). However, the protective effect of tin-protoporphyrin on bilirubin cerebellopathy (cerebellar hypoplasia) was less marked than expected. Possible implications of our results are discussed.

Animals↗
PubMed1987

Interaction of bilirubin with human erythrocyte membranes. Bilirubin binding to neuraminidase- and phospholipase-treated membranes.

Saturable bilirubin binding to human erythrocyte membranes was measured before and after digestion with neuraminidase and phospholipases. Neuraminidase-treated erythrocyte membranes did not show any change in their binding properties, indicating that gangliosides could be excluded as candidates for saturable bilirubin-binding sites on erythrocyte membranes. Although bilirubin-binding properties of the membranes did not change after phospholipase D digestion, either, phospholipase C treatment greatly enhanced bilirubin binding. Thus it is suggested that a negatively charged phosphoric acid moiety of phospholipids on the membrane surface may play a role to prevent a large amount of bilirubin from binding to the membranes. Further saturable bilirubin binding to inside-out sealed erythrocyte membrane vesicles showed values comparable with those of the right-side-out sealed membranes, suggesting that the bilirubin-binding sites may be distributed on both outer and inner surfaces of the membranes, or may exist in the membranes where bilirubin may be accessible from either side.

Bilirubin↗
PubMed1987

Age-dependent and selective binding of beta-bungarotoxin to GABAergic neurons in the rat cerebellum.

beta-Bungarotoxin (BuTx)-binding cells were immunocytochemically examined in the developing rat cerebellum. The tissue was incubated with BuTx and then immunostained with antiserum against its toxoid. On postnatal day 6, only Golgi cells were positive for immunoreaction. Immunoreactive Golgi cells were reduced in number on day 15 and disappeared on day 25. On day 15, Purkinje cells were strongly stained, while some basket and stellate cells stained weakly. On day 25 and in adult, basket and stellate cells were more immunoreactive than Purkinje cells. Thus, age-dependent and selective binding of BuTx was restricted to gamma-aminobutyric acid (GABA)ergic neurons.

Aging↗
PubMed1987

Purification of S-100a0 protein from rat kidney.

In order to corroborate our previous finding that kidney may contain a considerable amount of S-100-like antigen, concentrations of S-100a0, S-100a and S-100b antigens in rat kidney were determined by an enzyme immunoassay system, which can quantify these 3 forms of S-100 protein separately. It was revealed that rat kidney was rich in S-100a0 antigen, while it was the least occurring form in brain. To confirm the above results, S-100-like antigen in rat kidney was purified, and its physicochemical properties were compared with those of S-100 proteins of bovine brain. Electrophoretic mobility, calcium binding ability, apparent molecular weight, amino acid composition as well as elution profiles from butyl-Sepharose and anion exchange columns were similar to those of S-100a0 protein of bovine brain. These results indicate that S-100a0 protein is not a protein unique to brain, as had long been believed.

Amino Acids↗
PubMed1987

Purification of specific antibody against aspartate and immunocytochemical localization of aspartergic neurons in the rat brain.

The distribution of L-aspartate known as a putative excitatory neurotransmitter in the central nervous system was investigated immunocytochemically in the rat brain. Anti-aspartate antiserum was raised in rabbits using L-aspartate covalently conjugated to rabbit serum albumin with glutaraldehyde as the immunogen and was found to be cross-reactive with an L-glutamate conjugate. Monospecific anti-L-aspartate antibody was successfully purified using affinity gels coupled with several amino acids including L-aspartate and L-glutamate and with the L-glutamate conjugate. Putative aspartergic neurons were generally immunoreactive to the purified antibody, but epithelia of the choroid plexus were also stained. These results show that the antibody is a useful tool for the immunocytochemical demonstration of possible aspartergic neurons in the central nervous system, although the immunochemical expression of L-aspartate not used as a neurotransmitter must be taken into consideration.

Animals↗
PubMed1987

High-molecular-weight cadmium-binding proteins in rat liver cytosol: isolation and partial characterization of an approximately 50-kDa protein.

The time-dependent changes in the chromatographic pattern of subcutaneously injected cadmium associated with non-metallothionein cadmium-binding proteins were studied in the rat liver cytosol. Prior to the induction of cadmium-thionein (less than 3 h), cadmium appeared in three major peaks (P-1 with the void volume, P-2 and P-3) on Sephacryl S-300 column chromatography. Accompanied with the emergence of apo-metallothionein (about 3 h after administration), the amount of P-3 decreased and instead a cadmium-thionein peak (P-4) increased. Ion-exchange chromatography of P-3 with a combination of CM and DEAE Bio-Gel columns showed the existence of three major cadmium-binding proteins with molecular sizes of 46 kDa (in the CM Bio-Gel column eluate), 50 kDa (in the DEAE Bio-Gel column eluate), and 41 kDa (in the non-adsorbed fraction). The cadmium-binding protein in the CM Bio-Gel column eluate was purified to apparent homogeneity. The purified protein (CM-CdP) was 47 or 53 kDa in molecular size as determined by SDS-polyacrylamide gel electrophoresis or gel filtration chromatography, respectively. The apparent dissociation constant and maximum binding for cadmium were about 1 microM and 1 mol of the metal/mol of protein, respectively. The isoelectric point was estimated to be 8.8. The amino acid composition showed that the protein was relatively rich in glutamyl (including its amide) and alanyl residues. The N-terminal amino acid sequence was determined as Ala-Pro-Ile-Ala-Gly-Lys-Lys-Ala-Lys-Ala-Gly-Ile-Leu-Leu-Gly-. In-vitro experiments revealed that cadmium bound to CM-CdP could be easily transferred to apo-metallothionein, confirming that the affinity for the metal of the former protein was lower than that of the latter.

Amino Acid Sequence↗