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Y Hotta

Publications and source records attributed to Y Hotta.

At least 163 records · Page 9Linked to original sources
PubMed1990

A 49-kilodalton phosphoprotein in the Drosophila photoreceptor is an arrestin homolog.

The gene encoding the 49-kilodalton protein that undergoes light-induced phosphorylation in the Drosophila photoreceptor has been isolated and characterized. The encoded protein has 401 amino acid residues and a molecular mass of 44,972 daltons, and it shares approximately 42 percent amino acid sequence identity with arrestin (S-antigen), which has been proposed to quench the light-induced cascade of guanosine 3',5'-monophosphate hydrolysis in vertebrate photoreceptors. Unlike the 49-kilodalton protein, however, arrestin, which appears to bind to phosphorylated rhodopsin, has not itself been reported to undergo phosphorylation. In vitro, Ca2+ was the only agent found that would stimulate the phosphorylation of the 49-kilodalton protein. The phosphorylation of this arrestin-like protein in vivo may therefore be triggered by a Ca2+ signal that is likely to be regulated by light-activated phosphoinositide-specific phospholipase C.

Amino Acid Sequence↗
PubMed1990

Giant Drosophila neurons differentiated from cytokinesis-arrested embryonic neuroblasts.

The relative contributions of the intrinsic and extrinsic factors in determining neuronal differentiation are not fully understood yet. We found that isolated neuroblasts from Drosophila gastrulae were able to differentiate neuron-specific properties in culture even when cell divisions were inhibited. The resultant giant multinucleated neurons displayed thickened neurites with a variety of distinct branching patterns. Neuronal antigens were expressed as in normal cultured neurons, and action potentials could be evoked by current injection within two days after plating. These results indicate that the factors for initiating specific differentiation programs for basic neuronal form and function are present in a neuroblast already. The cells of increased sizes in this culture system are more accessible to physiological and cell biological analyses and could facilitate future studies of the Drosophila nervous system.

Action Potentials↗
PubMed1990

ATP-independent strand transfer protein from murine spermatocytes, spermatids, and spermatozoa.

A protein-catalyzing D-loop formation is present in murine spermatocytes, spermatids, and spermatozoa, but is not found in somatic tissue or in premeiotic cells of the germline. Unlike the Escherichia coli RecA protein and the meiotic rec protein (m-rec) previously described, D-loop formation by this protein (referred to as "mAi-rec") does not require ATP. The meiotic profile of mAi-rec activity is only partly similar to that of m-rec. Like m-rec, it rises steeply during early prophase and reaches a peak at pachytene. Unlike m-rec, its activity remains high during the postmeiotic phase of spermatid development and is prominent in immunochemically stained spermatozoa. A polyclonal antibody to E. coli RecA reacts with mAi-rec and inhibits its activity. No such reaction occurs with m-rec protein. The extent of sequence homology between E. coli RecA and murine mAi-rec is highly limited; none of the several monoclonal antibodies tested reacted with mAi-rec.

Adenosine Triphosphate↗
PubMed1990

Regulation of Drosophila myosin ATPase activity by phosphorylation of myosin light chains--II. Flightless mfd- fly.

1. The Ca2(+)-activated and Mg2+ actin-activated myosin ATPase activities of flightless mfd- mutant Drosophila flight muscle myosin were one-half and one-third of those of the wild-type fly muscle myosin, respectively. 2. In the two-dimensional gel electrophoresis, the spots corresponding to phosphorylated myosin light chains, Lfl and Ltl, were hardly detected in mfd- mutant myosin. 3. These results support not only the conclusion that phosphorylation of myosin light chains regulates Drosophila myosin ATPase activity but also the assumption that the phosphorylation of myosin light chains is directly involved in flight function of the Drosophila fly.

Animals↗
PubMed1990

Actin with tumor-related mutation is antimorphic in Drosophila muscle: two distinct modes of myofibrillar disruption by antimorphic actins.

A mutant beta-actin with an amino acid substitution from Gly-245 to Asp has been shown to be related to tumorigenic transformation of a human fibroblast cell line (Leavitt, J. et al. (1987) Mol. Cell. Biol. 7, 2467-2476). To examine the effects of this mutation, we artificially introduced the same amino acid change into the Act88F actin gene of Drosophila melanogaster. The gene (Act88FGD245) was inserted in the Drosophila genome to make transgenic adult flies which synthesize the mutant actin in the indirect flight muscles. The mutant actin was found to be antimorphic with regard to flight and also to cause myofibrillar disruption in transformants even in the presence of two normal alleles. It was initially incorporated into myofibrils and later induced their degeneration from center to periphery. This mode of myofibrillar disruption is distinct from that of previously reported Act88F mutations, where defects are found only in the peripheral region of myofibrils. This indicates that actin functions are altered differently in the two classes of antimorphic mutations.

Actins↗
PubMed1990

Purification of DNA ligases from mouse testis and their behavior during meiosis.

Two types of DNA ligase, I and II, have been purified approximately 4,000-fold from mouse testes and 500-fold from nuclei of mouse spermatocytes. DNA ligase I and II consisted of single polypeptides with molecular weights of 95,000 and 65,000, respectively, according to the estimation by SDS-polyacrylamide gel electrophoresis and the AMP-binding assay. Ligase activities were higher in premeiotic spermatogonia and spermatocytes than those in liver and bone marrow cells. Moreover, DNA ligase II showed rapid increase during meiotic prophase and a decrease in round spermatids. Since this behavior of DNA ligase II is consistent with that of m-rec and DNA polymerase beta, both of which have been shown to be involved in DNA recombination in meiotic cells, DNA ligase II might be an enzyme which works at the final step of meiotic recombination reaction.

Adenosine Monophosphate↗
PubMed1990

[Morphological study using Moiré patterns on the dental arch and palate of Japanese with normal occlusion].

Studies of the dental arch and the palate have progressed from two-dimensional to three-dimensional analysis. However, many points still need clarification in this field even though the fundamental structure of arch and palate have a common function in connection with mastication. The present study, using three-dimensional measurements of the palate, endeavors to clarify the differences in palatal morphology as these are affected by the changes in the dental arch morphology. Casts were made using 62 males between the ages of 20-25. Only those casts which were considered to represent normal occlusion of the upper dentition were selected for use in this study. Photographs of the casts were studied using the Moiré topographical method with a grid of 0.5 mm. Seven points were selected for measurement purposes (figs. 2 and 3). Based on these points, the seven distances were used in this study. Measuring of the palate was performed in the following manner: lines were drawn radially from the origin (F) to the most lingually protrusive point on the gingiva of each of the 12 teeth from the right first molar round to the first molar on the left side. The points of mergence by these lines with every second line (2 mm deep) of the Moiré pattern were fed into a digitizer. The results were summarized as follows: 1. The anterior, posterior and complete arch chords were 23.73 mm, 19.93 mm and 41.68 mm respectively. The anterior, posterior and complete arch lengths were 14.14 mm, 19.98 mm and 34.12 mm respectively. The anterior and posterior arch widths were 38.02 mm and 50.73 mm respectively. 2. As for the correlation coefficients, there was a correlation between the lengths and widths of the anterior portion, but not for the posterior. 3. The length from the origin (F) to the papilla incisiva was 25.4 mm, and the depth of the palate at the origin (F) was 13.4 mm on the average. 4. In the vertical cross section of the palate for each tooth, the slope line of the palate gradually increased from the incisors to the first molars, but the amount varied between the anterior and posterior portions of the maxillary dentition. Also, the line of the palate converged at one point about 11 mm from the origin (between lines 9 and 11 on the Moiré pattern). 5. In the horizontal cross section of the palate, the shape of the palate was almost semi-circular in the range of Moiré lines 9-11 with the origin at the center.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗
PubMed1990

Analysis of photoreceptor membrane turnover in a Drosophila visual mutant, rdgA, by electron microscope autoradiography.

A Drosophila visual mutant, rdgA, has photoreceptor cells whose rhabdomeres degenerate in several days after eclosion. Incorporations of 3H-amino acids, and 3H-mannose and 3H-glucosamine residues into the photoreceptive membranes were studied in newly emerged rdgA mutant flies by electron microscope autoradiography. The amount of 3H-amino acids incorporated in rdgA rhabdomeres at 3 hr after the injection was about 50%, and that of 3H-sugar residues was about 20% of normal. Together with our previous finding that degradative activity is low in rdgA cell bodies at this time, these data indicate that the supply of photoreceptive membrane proteins is defective in rdgA.

Amino Acids↗
PubMed1989

Point mutation affecting processing of the ornithine aminotransferase precursor protein in gyrate atrophy.

A generalized deficiency of the mitochondrial enzyme, ornithine aminotransferase (OAT) is the inborn error in gyrate atrophy, an autosomal recessive degenerative disease of the choroid and retina of the eye that leads to blindness. Southern analysis, using the OAT cDNA probe, of the OAT gene in a gyrate atrophy patient whose level of OAT protein is markedly decreased indicated the functional gene to be grossly intact. Northern analysis of his OAT mRNA demonstrated only half the normal level of OAT message, suggesting expression of only one of the two alleles of the OAT gene. A functional assay of the expressed OAT mRNA by in vitro translation and immunoprecipitation with anti-human OAT antibody indicated synthesis of an OAT protein from the message. The expressed message was cloned and sequenced and was shown to contain a single base change from C to T, resulting in an amino acid codon change from CAT (histidine) to TAT (tyrosine) at position 319 in the translated OAT protein. The mutant and normal OAT precursors were synthesized using transcriptional expression clones of OAT and in vitro translation of the expressed mRNA and tested in an in vitro mitochondrial transport/processing system. The results indicate that the mutant OAT precursor from the gyrate atrophy patient can be transported to the mitochondria but is minimally processed there, which would lead to degradation of the labile precursor and loss of OAT activity as phenotypically observed.

Adult↗
PubMed1989

Diacylglycerol kinase defect in a Drosophila retinal degeneration mutant rdgA.

The Drosophila visual mutant rdgA is known to show age-dependent retinal degeneration with defective diacylglycerol (DG) kinase activity. In this study we examined DG kinase activity of several visual mutants and found that only rdgA mutant eyes showed the lack of DG kinase activity in a gene dosage-dependent manner. The enzyme activity is already absent at the time of eclosion from pupal case when the degeneration is not yet apparent. To examine whether rdgA gene dosage effect holds for other enzymes related to the phosphatidylinositol turnover, phospholipase C was analyzed which did not show any gene dosage effect. Therefore, it is strongly suggested that rdgA gene correlates closely with DG kinase activity, and the defect of DG kinase activity is a primary cause of retinal degeneration in rdgA mutant.

Animals↗
PubMed1989

Structure of the subrhabdomeric cisternae in the photoreceptor cells of Drosophila melanogaster.

The structure of subrhabdomeric cisternae (SRC) and related structures in the photoreceptor cells (retinular cells) of Drosophila melanogaster in normal flies and visual mutants were compared by electron microscopic observation of semithin sections of osmium-impregnated specimens. The three-dimensional organization of SRC and the other cell organelles was demonstrated by stereoscopy. Both light- and dark-adapted normal retinular cells contained elaborate networks of anastomosing tubules of SRC immediately beneath the rhabdomeres. Tubules connecting the SRC and rough endoplasmic reticulum were frequently seen. The SRC were absent from the retinular cells of rdgAKS60 whose rhabdomeres degenerate gradually after eclosion. Instead, numerous smooth vesicles were observed in the subrhabdomeric regions. In rdgBEE170, in which rhabdomere degeneration is light dependent, the SRC appeared normal in the dark-adapted flies. But their SRC gradually disintegrated after exposure to light. In norpASB37, whose rhabdomeres are small but do not degenerate, SRC appeared normal. These results suggest that the SRC is a significant structure for the maintenance of the structure of photoreceptive membrane in the retinular cells of Drosophila.

Animals↗
PubMed1989

Difference in the expression level of DNA polymerase beta among mouse tissues: high expression in the pachytene spermatocyte.

The expression level of DNA polymerase beta was determined in various mouse tissues. Northern blot hybridization analysis using rat cDNA as a probe revealed that the mRNA of about 1.5 kb for this enzyme is present in all kinds of tissues examined, but its content widely varies among tissues; the most abundant DNA polymerase beta mRNA was present in the testis, which was followed by brain, thymus, and spleen. The mRNA content was low in heart, kidney, and liver. In testis and brain, two minor species of transcripts of 3.3 and 6.2 kb were detected in addition to that of 1.5 kb. DNA polymerase beta activities in these tissues were closely correlated with the mRNA content, indicating that the expression of this enzyme is mainly regulated by the level of the mRNA. A survey of DNA polymerase beta mRNA levels in the testes at successive postnatal developmental stages and in isolated spermatogenic cells indicated that DNA polymerase beta mRNA was most abundant in spermatocytes at early pachytene. Since meiotic recombination occurs in this period, DNA polymerase beta may be involved in the repair-type DNA synthesis associated with the recombination process.

Animals↗
PubMed1989

Pharmacological characterization of sodium channels in the primary culture of individual Drosophila embryos: neurons of a mutant deficient in a putative sodium channel gene.

Sodium channels in Drosophila embryonic neurons were characterized pharmacologically in the primary culture of individual gastrulae. In normal cultures, presence of sodium channels was demonstrated by neuronal degeneration in the presence of veratridine and ouabain, which was inhibited by tetrodotoxin. Embryonic neurons of Df(2R)M-c33a homozygotes that lack a putative sodium channel gene at 60E region showed normal neurotoxin sensitivity. Therefore, sodium channel genes other than 60E must be functional at this developmental stage. We also examined para(ts1) and nap(ts) mutants and found that they were also sensitive to the neurotoxins. To determine the genotypes of single embryo cultures by histochemical staining, we utilized a special chromosome bearing a hsp70-lacZ fusion gene. The lacZ expression in the culture was studied in detail.

Animals↗
PubMed1989

Plant saponins can affect DNA recombination in cultured mammalian cells.

Some plant extracts and products are known to affect mammalian cells, tissues and organisms as they contain a toxic substance or a metabolic stimulant. Our biochemical investigations revealed that some plant saponins can increase the cellular DNA repair activity and the general recombinase activity measured by in vitro assay (1). In the experiments described here, HeLa cells were cultured for several days with plant saponins or flavonoids and analyzed to measure i) recombination activity of the cell extract by induction of Tcr colonies from two mutant DNAs (mutants 1 and 2, which are both tetracycline sensitive) after transformation into E. coli recA-, and ii) repair synthesis of nuclear DNA followed by incorporation of 3H-thymidine. Saikosaponins a, b1, d, ginsenosides Rb1, Re, Rh and flavonoid baicalin caused a significant stimulation of intermolecular recombination. It is worth noting that none of the plant saponins and flavonoids had any inhibitory or toxic effect at concentrations less than 25 micrograms/ml in the culture media.

Animals↗
PubMed1989

Studies on the chemical modification of monensin. II. Measurement of sodium ion permeability of monensylamino acids using sodium-23 nuclear magnetic resonance spectroscopy.

A technique to assay Na+ ions in cells is presented. Intracellular and extracellular Na+ ions in a suspension of guinea pig erythrocytes were conveniently determined by using sodium-23 nuclear magnetic resonance (23Na-NMR), in combination with two anionic shift reagents: Dy(TTHA)3- and Dy(PPPi)2(7-). Monensin (1), monensylalanine (2b), monensylserine (2c), and monensylphenylalanine (2d) induced large increases of intracellular Na+ ion concentration ([Nain]), while monensylglycine (2a), monensyltyrosine (2e), monensylaspartic acid (2f), and monensylglutamic acid (2g) showed slight increases. The values of initial increasing rate (Vi) of 2a-g were much smaller than that of 1. This fact was probably due to the lower lipophilicity of 2a-g than 1, because a good correlation was observed between Vi and Rm50 values of 1 and 2a-g. This lower lipophilicity is a consequence of conformational differences between 1 and 2a-g.

Amino Acids↗
PubMed1989

[Clinical evaluation of synthetic hydroxyapatite implants for human periodontal osseous defects. 6 cases studied more than 3 years after the surgery].

We applied of granular hydroxyapatite HAP to periodontal bone defects, for the purpose of restoring the periodontal tissue destroyed by periodontal disease, the subjects were 6 patients (6 regions) with chronic marginal periodontitis who had passed more than 3 years since HAP implantation. After routine periodontal initial treatment, a flap operation was performed, and granular HAP, or a combination of granular and block-like HAP, was implanted into the periodontal bone defect. Clinical parameters were probing depth, clinical attachment level, clinical gingival margin, residual HAP volume and X-ray before operation, at 3, 6, 12 months, and 2, 3, and 4, 6 years after the operation. The results revealed that all measured values showed good results for both granular HAP and the combination of granular and block-like HAP after implantation, and that all cases showed stable results from 12 months on. No remarkable difference was noted between the implanted regions.

Bone Resorption↗