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Biomedical subjects

S Nishibayashi

Publications and source records attributed to S Nishibayashi.

26 records · Page 2Linked to original sources

Alterations in the binding of the phosphodiesterase inhibitor, rolipram, after transient ischemia in the gerbil brain.

To determine ischemia-induced changes in phosphodiesterase (PDE), changes in the membranous binding sites of rolipram, a cAMP-selective PDE inhibitor, were examined in the gerbil brain following transient 5 min forebrain ischemia. Coinciding with the delayed neuronal death (DND) in the hippocampal CA1 region, affinities for cerebral rolipram bindings decreased on Day 4, when intrinsic cAMP, substrate for PDE, might increase. The number of rolipram binding sites was significantly reduced in the hippocampus Day 14, despite the lack of change on Day 4. This reduction in rolipram binding was in agreement with the previously reported late onset reduction of muscarinic receptors, progressing more slowly than DND. Slowly progressive mechanisms may be involved in the ischemia-induced reduction of the hippocampal rolipram binding sites which may be PDEs.

Analysis of Variance↗

Variance of ploidy in mitochondrial nucleus during spherulation in Physarum polycephalum.

In Physarum polycephalum, microplasmodia differentiated into spherules when cultures were aged for 8-10 days. Respiration rates of the microplasmodia decreased rapidly with ageing to a 90% decrease in oxygen consumption over 9 days. We studied this phenomena by isolating and characterizing mitochondria from microplasmodia and spherules at different stages of spherulation. Oxygen uptake by the isolated mitochondria decreased with spherulation. Morphological and biochemical analyses showed that mitochondrial differentiation to inactive state was characterized by a decrease not only in dimension but also of content (DNA, RNA and protein). Diminutive mitochondria contained small particle-shaped mitochondrial nuclei. The DNA content, measured by microscopic fluorometry, was about 1.15 and 0.58 X 10(-10) g, which corresponded to about 16 and 8 genome copies, respectively (e.g., 32 genome copies per mitochondrion at mitochondrial G1). Restriction endonuclease analysis showed that the physical structure and methylation pattern of the mtDNA had not changed although the DNA content per mitochondrion had decreased remarkably with spherulation. This showed that changes in the ploidy level of the mitochondrial nucleus during spherulation were due to reduction in the number of whole mitochondrial genomes.

Cell Fractionation↗

Epifluorescent microscopic evidence for maternal inheritance of chloroplast DNA.

Maternal inheritance of chloroplast genes occurs in the isogamous green alga Chlamydomonas reinhardii. It has been shown using biochemical techniques that the chloroplast DNA of male origin is preferentially lost by 6 h after mating. DNAs in the chloroplast are organized by proteins into about 10 chloroplast nucleoids. Therefore, if chloroplast DNA in zygotes is preferentially destroyed, the disappearance of chloroplast nucleoids from male gametes should be observable during zygote formation by high resolution epifluorescent microscopy. Here we present the first fluorescent microscopic evidence that in C. reinhardii, about eight chloroplast nucleoids from the male parent disappear during the first 40-50 min after mating, while those from the female parent persist and finally fuse together to form one large chloroplast nucleoid.

Cell Fusion↗

In vitro nucleation of microtubules from microtubule-organizing center prepared from cellular slime mold.

Nucleus associated bodies (NABs) were isolated from Dictyostelium discoideum or Dictyostelium mucoroides and their ability to nucleate microtubules in vitro was examined. NABs were localized at the tapered ends of the nuclei and released from lysed cells in complex with the nuclei. Microtubules radiating from the NAB could also be isolated with the complex under microtubule stabilizing conditions. The ultrastructure of the isolated NAB showed it to be composed of a core structure surrounded by an amorphous matrix. The ability of isolated NABs to nucleate microtubules in vitro was demonstrated by incubation with exogenous brain microtubule protein. Microtubule assembly was easily visualized by dark-field or immunofluorescence microscopy. Polymerization of microtubules seemed to be initiated not from the core structure but from the surrounding matrix. The number of microtubules polymerized from the NAB was directly counted in whole-mount preparations by electron microscopy, which provided a quantitative assay for the NAB activity. The nucleating activity of NAB was quite unstable and its half-life was calculated as about 5 hours. The activity was sensitive to protease digestion and was also temperature sensitive but could be stabilized by addition of glycerol or storage at - 80 degrees C or in liquid nitrogen. These characteristics are analogous to those of the centrosomes in cultured mammalian cells and a possible explanation of their similarity is discussed.

Cell Fractionation↗

Variance of ploidy in Candida albicans.

Determination of ploidy was performed on isolates of Candida albicans from clinical sources by measuring nuclear DNA content with fluorescent microscope photometry. By this criterion and UV irradiation survival experiments, haploid, diploid, and tetraploid strains were identified in this organism. The dimensions of nucleus-associated organelles (equivalent to spindle pole bodies) in these strains increased as a function of ploidy number.

Candida albicans↗

Tubulin and actin mRNAs in the young-adult and the aged rat brain: effects of repeated administration with bifemelane hydrochloride.

In an attempt to identify the age-dependent changes in the potential synthesis of cytoskeletal proteins, we investigated changes in messenger RNA (mRNA) of alpha-tubulin and beta-actin in the young-adult and the aged rat brain using Northern blot analysis. alpha-Tubulin mRNA levels in the frontal cortex and hippocampus, and beta-actin mRNA levels in the hippocampus were significantly decreased in the aged rat brain. Age-dependent decreases in these mRNAs may be related to the neuronal dysfunction associated with aging, in addition to the reduction of several kinds of receptors previously reported. Repeated administration of bifemelane hydrochloride (4-(2-benzylphenoxy)-N-methylbutylamine hydrochloride) for 14 days increased the levels of beta-actin mRNA in the frontal cortex and the striatum of both young-adult and aged rats, although the effect of bifemelane treatment was smaller and not significant in the aged group. These results suggest that bifemelane treatment may enhance the synthesis of cytoskeletal protein and promote neural plasticity by inducing neurite growth or synapse formation.

Journal Article↗

Protective effects of pergolide on dopamine levels in the 6-hydroxydopamine-lesioned mouse brain.

Pergolide, along with bromocriptine and lisuride, is one of the most active dopamine receptor agonists. To determine whether or not pergolide protects against dopaminergic neuronal damage, via its activity on monoamine metabolism, we studied the effects of pergolide pretreatment on changes in monomaines and their metabolites in the mouse striatum after intracerebroventricular injection of 6-hydroxydopamine with pretreatment of desipramine. After intracerebroventricular administration of 6-hydroxydopamine (40 micrograms) in mice, the levels of dopamine and its metabolites (DOPAC, HVA) in the striatum rapidly decreased to 49%, 29% and 68%, respectively, of the naive controls at week 1 but then gradually recovered to control levels at weeks 2 and 4. Repeated pretreatment with pergolide (0.5 mg/kg, i.p.) for 7 days before administration of 6-hydroxydopamine, almost completely protected against reduction in striatal dopamine and its metabolites 1 week after injection of 6-hydroxydopamine. Therefore, pergolide could normalize the decreased dopamine synthesis or storage, and has a neuroprotective effect against dopaminergic dysfunction induced by the neurotoxin, 6-hydroxydopamine. Although we found that pergolide did not show radical scavenging activity in an in vitro system that generated hydroxyl radicals, it has been reported in vivo that pergolide treatment may induce Cu/Zn superoxide dismutase in the rat striatum. Considering these findings, pergolide may well be protective to dopaminergic neurons, largely because of its effects on presynaptic autoreceptors and on its induction of Cu/Zn superoxide dismutase. Further research on the neuroprotective effects of pergolide in Parkinson disease models, by injection of 6-hydroxydopamine, is needed to clarify its mechanism of action on dopaminergic indices.

3,4-Dihydroxyphenylacetic Acid↗