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R Semba

Publications and source records attributed to R Semba.

At least 55 records · Page 3Linked to original sources
PubMed1990

Appearance of parvalbumin-specific immunoreactivity in the cerebral cortex and hippocampus of the developing rat and gerbil brain.

Developmental changes in the distribution of parvalbumin-specific immunoreactivity in the brain, in particular in the cerebral cortex and hippocampus, were followed immunohistochemically in two different species, the rat and the Mongolian gerbil (Meriones unguiculatus) using an antibody raised against for rat parvalbumin. The gerbil is known to develop its auditory and visual capacity later than rat. In both the rat and gerbil, parvalbumin-specific immunoreactivity appeared after birth in both the cerebral cortex and hippocampus. The timing of the development of expression of parvalbumin varied among different parts of the cerebral cortex. The parietal cortex showed evidence of the earliest expression of parvalbumin whilst the occipital and temporal cortices expressed parvalbumin at a later stage of a development. This feature was common to both the rat and gerbil but occurred at a relatively later stage in the gerbil. The profile of the distribution of parvalbumin in the brain of the developing and adult gerbil was similar to that of the rat, but there were some differences. The frequency of bead-like structures on the dendrites of the parvalbumin-positive cells in the CA1 region of the hippocampus was markedly lower in the gerbil; instead, straight non-beaded fibers which ran vertically into the pyramidal layer were stained. Parvalbumin-positive fibers were also found in the cerebral cortex of the gerbil.

Animals↗
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

Identification of lung major GTP-binding protein as Gi2 and its distribution in various rat tissues determined by immunoassay.

Antisera were raised in rabbits against the 40-kDa alpha subunit of bovine lung GTP-binding protein, which were identified as the alpha subunit of Gi2 (Gi2 alpha) by the analysis of the partial amino acid sequence. Antibodies were purified with a Gi2 alpha-coupled Sepharose column and then were passed through a Gi1 alpha-coupled Sepharose column to remove antibodies reactive also with 41-kDa alpha. Purified antibodies reacted with Gi2 alpha, but not with Gi1 alpha, Gi3 alpha, or Go alpha in an immunoblot assay. A sensitive enzyme immunoassay method for the quantification of Gi2 alpha was developed by using these purified antibodies. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody Fab' fragments labeled with beta-D-galactosidase from Escherichia coli. The minimal detection limit of the assay was 1 fmol, or 40 pg. Samples from various tissues were solubilized with 2% sodium cholate and 1 M NaCl, and the concentrations of Gi2 alpha were determined. Gi2 alpha was detected in all the tissues examined in the rat. The highest concentration was found in platelets and leukocytes when the data were expressed as picomoles per milligram of protein. The spleen, lung, and cerebral cortex contained relatively high levels of Gi2 alpha. In the bovine brain, Gi2 alpha was distributed almost uniformly among the various regions. The concentrations of Gi2 alpha were constant in the rat brain throughout ontogenic development, in contrast with those of Go alpha which were markedly increased with age.

Amino Acid Sequence↗
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↗
PubMed1989

Highly sensitive enzyme immunoassay for bovine chromogranin A: application for studies of regional distribution in bovine central nervous system.

Bovine chromogranin A (CGA) was purified by three steps of column chromatography to a single-band purity in sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The antibody against this preparation was purified by a CGA-coupled Sepharose column, and F(ab')2 and Fab' of the antibody IgG were prepared by enzymatic digestion. An enzyme immunoassay (EIA) system was developed with the F(ab')2 immobilized on polystyrene balls and the Fab' labeled with beta-D-galactosidase. The EIA was able to detect 1 pg of CGA and was three to four orders more sensitive compared with any radioimmunoassay systems hitherto reported. Several neural acidic proteins (dopamine beta-hydroxylase, neuron specific enolase, S-100a protein, and brain-type creatine kinase) showed no cross-reaction. Using this EIA, CGA was detected in all regions of bovine central nervous system. CGA concentrations were within a narrow range, being lowest in the cerebellar white matter and highest in the putamen (17.3 and 78.1 ng/mg protein, respectively). The concentrations were extremely low compared to the concentration in the adrenal medulla (205,000 ng/mg protein). The highly sensitive EIA system should be useful for studies of materials containing very small amounts of CGA.

Animals↗
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

Ontogeny of the GTP-binding protein Go in rat brain and heart.

We determined the ontogeny of the GTP-binding protein Go in rat brain and heart by employing highly sensitive enzyme immunoassay methods. In the brain, the alpha subunit of Go (Go alpha) gradually increased and reached adult levels approximately 20 and 30 days after birth in cerebral cortex and cerebellum, respectively. Concentrations of beta subunits, which were also quantified by the immunoassay, were almost equal to those of Go alpha in the brain of rats younger than 10 days, but were higher than those of Go alpha after 10 days. These results suggest that late development of GTP-binding proteins other than Go. Go alpha was immunohistochemically positive in neuropils and negative in cell bodies at any age tested. In the heart, the concentrations of Go alpha increased up to several times of the adult level just after birth, and then gradually decreased after the 20th postnatal day. The level of Go alpha in the liver, however, was very low and constant throughout ontogenic development. An immunohistochemical study indicated that Go alpha was positive in the cardiac muscle of young rat, but negative in that of adult rat. These results indicate that Go alpha exists in cells other than those of nervous tissues and neuroendocrine cells in some periods of ontogenic development.

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

Go, a GTP-binding protein: immunochemical and immunohistochemical localization in the rat.

The tissue and cellular distribution of a GTP-binding protein, Go, was investigated in the rat by immunochemical and immunohistochemical methods. Because the specific antibody for the alpha subunit of bovine Go (Go alpha) cross-reacted with rat Go alpha, an enzyme immunoassay method developed for bovine Go alpha was applied for measuring the tissue concentration of Go alpha in the rat. Go alpha was detected in all tissues examined except blood cells. The concentration of Go alpha was highest in the CNS (approximately 7.7 and 4.4 nmol/g in the cerebrum and cerebellum, respectively), followed by the pituitary gland and sciatic nerve. Among the other peripheral tissues, relatively high concentrations of Go alpha were observed in the urinary bladder, stomach, and intestines; however, these values were less than 2% of the concentration in the cerebrum. Go alpha in the intestine was located mostly in the muscle layer. Immunohistochemical study showed that Go alpha was associated mostly with the neural elements but not with cells particular to each peripheral organ. Go alpha was also present in the membranes of neuroendocrine cells, including glandular cells in the anterior lobe of the pituitary gland, chromaffin cells in the medulla of the adrenal gland, islets cells in the pancreas, and parafollicular cells in the thyroid. These results indicate that Go is localized exclusively in the nervous tissues and neuroendocrine cells.

Animals↗
PubMed1988

Postsurgical epidural hematoma reproducing a lumbar radiculopathy.

A 24-year-old man presented to the spine service with an epidural hematoma three weeks after a successful decompressive laminectomy. Recurrence of the symptoms correlated with the repeat operative findings of a tenting hematoma over the area of the annular window. On decompression, the S1 nerve root relaxed in the L5-S1 gutter, effectively controlling the patient's symptoms and resulting in complete resolution of his pain. Radiographic and operative findings are discussed.

Adult↗
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↗