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T Nishimoto

Publications and source records attributed to T Nishimoto.

At least 181 records · Page 10Linked to original sources
PubMed1986

Distribution of light chains and ATPase activity of myosin in the atrioventricular conducting tissue of bovine heart.

The relations of the light chains of myosins of the atria, ventricles, and atrioventricular conducting tissue (specialized myocardial tissue) and the distribution of the light chains of myosin in different regions of the atrioventricular conducting tissue in bovine heart were examined. Two-dimensional gel electrophoresis showed that the atrial and ventricular myosins each had two light chains (LC1 and LC2). Ventricular LC1 differed from atrial LC1, but ventricular LC2 corresponded to atrial LC2. The specialized myocardial tissue myosin had three light chains (named here SL1, SL2, and SL3). SL1 comigrated with ventricular LC1, SL2 with atrial LC1, and SL3 with ventricular LC2 and atrial LC2. The compositions of the three light chains of myosins in various regions of the atrioventricular conducting tissue were determined by sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis. The percentage proportion of SL1 decreased in the order--atrioventricular node (AVN), right and left bundle branches (RLBB), His bundle (HIS), false tendon (FT) myosin; while the percentage proportion of SL2 decreased in the order--FT and HIS, RLBB, AVN myosin. The percentages of SL3 in these four regions were similar. The Ca2+-activated ATPase activity of myosin was highest in the AVN and lowest in the FT. The activities in the HIS and RLBB were intermediate between those in the AVN and FT. Thus, the composition of the light chains and the Ca2+-activated ATPase activity were different in various regions of the atrioventricular conducting tissue.

Adenosine Triphosphatases↗
PubMed1986

Improved purification of Aspergillus oryzae 1,2-alpha-mannosidase by using mannan gel affinity chromatography.

In order to facilitate the purification of 1,2-alpha-mannosidase from an enzyme product of Aspergillus oryzae, we have devised a rapid and simple procedure. A partially purified enzyme preparation obtained from the A. oryzae enzyme product, by means of ammonium sulfate fractionation followed by CM-Sephadex C-50 chromatography, was subjected to affinity chromatography with baker's yeast mannan gel as an adsorbent. 1,2-alpha-Mannosidase was retarded and well separated from the major protein peak on the affinity column. After a second affinity chromatography under the same conditions, 1,2-alpha-mannosidase was finally purified 7,500-fold with a 22.9% yield. The enzyme preparation thus obtained was quite suitable for the structural analysis of glycoconjugates.

Aspergillus↗
PubMed1986

Molecular cloning of a human gene that regulates chromosome condensation and is essential for cell proliferation.

The tsBN2 cell line, a temperature-sensitive (ts) mutant of baby hamster kidney cell line BHK21/13, seems to possess a mutation in the gene that controls initiation of chromosome condensation. At the nonpermissive temperature (39.5 degrees C), the chromatin of tsBN2 cells is prematurely condensed, and the cells die. Using tsBN2 cells as a recipient of DNA-mediated gene transfer, we investigated a human gene that is responsible for regulation of chromosome condensation and cell proliferation. We found that the human gene complementing the tsBN2 mutation resides in the area of the 40- to 50-kilobase HindIII fragment, derived from HeLa cells. Based on this finding, we initiated cloning of a human gene complementing the tsBN2 mutation. From lambda and cosmid libraries carrying partial digests of DNA from the secondary transformants, the 41.8-kilobase HindIII fragment containing the human DNA was isolated. The cloned human DNA was conserved in ts+ transformants through primary and secondary transfections. Two cosmid clones convert the ts- phenotype of tsBN2 cells to ts+ with more than 100 times a higher efficiency, compared with cases of transfection with total human DNA. Thus, the cloned DNA fragments contain an active human gene that complements the tsBN2 mutation.

Cell Cycle↗
PubMed1986

A technique for decalcification and demonstration of substance P-like immunoreactivity.

Substance P-like immunoreactivity (SPLI) was demonstrated in mouse spinal cord by an indirect immunofluorescence method after decalcification of the vertebra with a mixture of EDTA and Zamboni's fixative. SPLI was observed mainly in the gray matter of the spinal cord, especially the superficial layers of the dorsal horn; the distribution was the same as in the control spinal cord. No diffusion and depletion of SPLI were recognized after decalcification and no specific fluorescence was observed. The findings reported here indicate that decalcification with a mixture of EDTA and Zamboni's fixative is a useful method for examining SPLI in nervous tissue surrounded in situ by calcified tissues.

Animals↗
PubMed1985

Specific site of histone H3 phosphorylation related to the maintenance of premature chromosome condensation. Evidence for catalytically induced interchange of the subunits.

Previously we have found that histone H1 and H3 of tsBN2 cells showing premature chromosome condensation (PCC) at nonpermissive temperature (40.5 degrees C) were phosphorylated extensively as in mitotic cells (Ajiro, K., Nishimoto, T., and Takahashi, T. (1983) J. Biol. Chem. 258, 4534-4538). Under the influence of various chemicals, both the prevention of the PCC induction and the suppression of H3 phosphorylation occurred simultaneously, whereas H1 phosphorylation did not. At the minimum concentration for the inhibition of PCC induction, H1 phosphorylation remained at the control level, but H3 phosphorylation was completely suppressed. Tryptic peptide analysis revealed that the H3 phosphopeptide in PCC was single, and it was observed in the same position as in mitosis. The results suggest that specific site(s) of H3 phosphorylation related to the maintenance of a condensed state of chromatin.

Binding Sites↗
PubMed1985

Immunohistochemical observation on substance P in regenerating taste buds of the rat.

The present study was performed to investigate the relationship between substance P-positive (SP-positive) nerve fibers and regeneration of taste buds in the foliate papillae of the rat by means of immunohistochemistry. It was confirmed by neurotomy that taste buds in the foliate papillae of the rat were innervated mostly (90%) by the glossopharyngeal (IXth) nerve and partly (10%) by the chorda tympani. In this experiment, the IXth nerve was sectioned distal to the petrosal ganglion. The rats were sacrificed at various intervals from 20 days to 80 days after the operation. In the course of degeneration and disappearance of taste buds, both SP-positive fibers and taste buds disappeared completely from the posterior folds of the foliate papillae 7 days after the operation. Within 22 days, regenerated SP-positive fibers began to appear in the lamina propria, and following the penetration of the fibers into the epithelium, taste bud anlagen reappeared at the bottom of the trench, and in the posterior folds at 24 days. The process of new taste bud formation extended toward the apex of the trench and to the anterior folds, which seemed to follow the regeneration in the nerve trunk. Quantitative data showed a gradual increase in the number of taste buds and taste buds containing SP-positive fibers. These findings indicate that SP might have a role in regeneration of taste buds.

Animals↗
PubMed1985

The induction of chromosome condensation in tsBN2, a temperature-sensitive mutant of BHK21, inhibited by the calmodulin antagonist, W-7.

The induction of premature chromosome condensation (PCC) in tsBN2 cells, a temperature-sensitive (ts) mutant of BHK21/13 which shows PCC at the non-permissive temperature, was almost completely inhibited by 40 microM W-7, an antagonist of calmodulin. The mitotic phosphorylation of histone H1 and H3 was also inhibited by W-7. W-5, a chlorine-deficient analogue of W-7 and which interacts weakly with calmodulin, did not inhibit the induction of PCC, even at a dose of 80 microM. The content of calmodulin in tsBN2 cells was increased by a temperature shift to 40.5 degrees C. All these results suggested that calmodulin is required for the chromosome condensation.

Animals↗
PubMed1985

Phosphorylation of nonhistone proteins during premature chromosome condensation in a temperature-sensitive mutant, tsBN2.

In tsBN2 cells, a temperature-sensitive (ts) mutant of the BHK21 cell line, with a ts-defect in its regulatory system for chromosome condensation, antigens that react with mitotic specific mouse monoclonal antibody MPM-2 were produced when premature chromosome condensation (PCC) was induced by a temperature shift. The polypeptides of antigens recognized by MPM-2 in tsBN2 cells with PCC were identical to those of antigens in mitotic cells. These antigens appeared concomitantly with chromosome condensation, which suggests that these mitotic-specific antigens may be related to chromosome condensation. As the production of mitotic-specific antigens was inhibited by W-7, a specific and potent antagonist of calmodulin, calmodulin may function in the mitotic phosphorylation of nonhistone protein.

Animals↗
PubMed1985

Chromosomes of G2 arrested cells are easily analyzed by use of the 'tsBN2' mutation.

The tsBN2 cell line, a temperature sensitive (ts) mutant of the BHK21/13 cell line (Nishimoto, T. et al. Somatic Cell Genet. 4, 323-340, 1978) has a ts defect in its regulatory mechanism for the initiation of chromosome condensation, the so-called, premature chromosome condensation (PCC) being induced at a nonpermissive temperature (Nishimoto, T. et al. Cell 15, 475-483, 1978, Nishimoto, T. et al. J. Cell Physiol. 109, 299-308, 1981). Using the 'tsBN2' mutation, we analyzed chromosomes of tsBN2 cells arrested in the G2 phase with neocarzinostatin (NCS) and found considerable aberrations, such as gaps, breaks and double minutes-like fragmentation, in addition to a typical G2-chromosome (a long filamental chromosome made up of two chromatids). Our results provide evidences that the tsBN2 cell line can be used for examinations of the chromosomes of cells arrested in the G2 phase.

Animals↗
PubMed1985

Relationship between pelviureteral peristaltic frequency and urine flow change evoked by autonomic drug administration.

Parallel recording of renal pelvic pressure, ureteral electromyogram and urine volume was carried out in dogs given intravenous injections of noradrenaline, isoproterenol or acetylcholine. With noradrenaline, an increase in urine volume was observed coincidentally with an elevation of renal pelvic pressure and an increase in frequency of ureteral peristalsis. In response to isoproterenol, the urine volume diminished in harmony with flattening of pelvic pressure waves and the disappearance of ureteral peristalsis. Conversely, ureteral peristalsis increased in frequency immediately after the administration of acetylcholine, while the urine volume diminished.

Acetylcholine↗
PubMed1985

Changes in the ureteral peristaltic rate and the bolus volume in gradual and rapid urinary flow increase.

The electromyogram and bolus volume of ureteral peristalsis were measured during gradual and rapid urinary flow increase, using 16 mongrel dogs. When the urine flow was increased rapidly by administration of diuretics, the ureteral peristaltic rate always increased first, then the urine bolus volume increased and finally the ureter became a urinary column without forming the urine bolus. When the urine flow was increased gradually by intravenous drip infusion of saline solution, the ureteral peristaltic rate and urine bolus volume did not always increase in spite of urinary flow increase. The peristaltic rate and bolus volume helped each other for effective urinary transport, as both of them not only always increase but also one of them occasionally decreased.

Animals↗