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S Togashi

Publications and source records attributed to S Togashi.

At least 37 records · Page 2Linked to original sources

Developmental profile of P element transposition in Drosophila somatic cells.

Transposition of the P element during Drosophila ontogenesis was monitored. A modified P element was transposed by the P delta 2-3 transposase source. P elements inserted into the genome were cloned by the plasmid rescue at various developmental stages of the G1 hybrid to trace events in somatic cells. The transposed elements were directly counted by analyzing RFLP of genomic DNA fragments flanking the P elements. Transposition began from the late embryonic stage, but occurred rarely. Frequent transposition was observed from the late third instar to early pupal stage. From these results, transposition of the P element would appear to be affected by the developmental state of somatic host cells.

Animals↗

Anti-idiotypes against an anti-haloperidol antibody bind to sigma receptors.

Anti-idiotypic monoclonal antibodies that interact with the binding site of sigma receptors were generated. First, BALB/c mice were immunized with a haloperidol-bovine serum albumin conjugate, and monoclonal anti-haloperidol antibodies that recognize the piperidinyl moiety of haloperidol molecule were obtained. Second, for generation of anti-idiotypic antibodies, BALB/c mice were immunized with the anti-haloperidol monoclonal antibodies coupled to keyhole limpet hemocyanin. Anti-idiotypic antisera and three hybridomas secreting anti-idiotypic monoclonal antibodies were obtained. All of them were shown to inhibit [3H]haloperidol binding to the anti-haloperidol antibodies. The anti-idiotypes were potent in displacing the binding of [3H]haloperidol to rat brain sigma receptors. Furthermore, they significantly immunoprecipitated the sigma receptors from a detergent-solubilized preparation. These findings demonstrate the generation of anti-idiotypic monoclonal antibodies specifically interacting with membrane-bound and solubilized sigma receptors.

Animals↗

Mutation of twins encoding a regulator of protein phosphatase 2A leads to pattern duplication in Drosophila imaginal discs.

The Drosophila gene twins was identified through a P-element-induced mutation that caused overgrowth in posterior regions of the wing imaginal disc. Analyses using position-specific markers showed that the inactivation of this locus induced the formation of extra wing blade anlagen in the posterior compartment of the disc. The duplication was mirror symmetrical, and the line of the symmetry did not correspond to any of the known compartment borders. We isolated the twins gene and found that it encoded one of the regulatory subunits of protein phosphatase 2A (PP2A). These results suggest a novel aspect of physiological roles of protein dephosphorylation; that is, the control of PP2A activity is crucial for specification of tissue patterns.

Amino Acid Sequence↗

Structure of the Drosophila melanogaster gene encoding cyclin A.

A P element (PE)-induced Drosophila melanogaster mutation, hari, affects the formation of the bristle mechanosensory organ in the adult fly. In this mutation, the site of PE insertion is in the first intron of the gene (CycA) encoding cyclin A (CycA). In order to analyze the hari mutant at the molecular level, we cloned and sequenced the cDNA and genomic DNA encoding CycA. CycA has seven exons and six introns, and its transcription unit spans 6 kb. All exon-intron junctions are compatible with the GT/AG consensus. Results of primer extension analysis and RNase protection assay indicate that CycA has major and minor transcription start points (tsp). To our knowledge, this is the first report on the CycA genomic sequence from a multicellular organism.

Amino Acid Sequence↗

Efficient amplification of Drosophila simulans copia directed by high-level reverse transcriptase activity associated with copia virus-like particles.

The number of retrotransposon copia per genome in Drosophila melanogaster cultured cells is two to three times higher than that in D. melanogaster embryo cells. Here, we have found that the genome of the related species, Drosophila simulans, contains in cultured cells more efficiently amplified copia DNA (approximately ten fold). Furthermore, we analyzed copia virus-like particles (VLPs) prepared from D. melanogaster and D. simulans cultured cells, which contain copia RNA and reverse transcriptase (RT) activity, and thus, play a major role in copia replication. The RT activity associated with the D. simulans VLPs was 25 times higher than that associated with the D. melanogaster VLPs. Taken together with the fact that copia is believed to transpose through an RNA intermediate, these results suggest that the amplification of copia DNA should relate to copia RNA-mediated transposition, and the higher RT activity associated with the D. simulans VLPs would lead to the efficient amplification of copia DNA. In a comparison between D. melanogaster and D. simulans copia nucleotide (nt) sequences, five nt substitutions, which cause the respective amino acid changes, were found in the copia RT-coding region. Polymerase chain reaction direct sequencing showed that these five substitutions are the vast majority in each Drosophila species. The substitutions, therefore, may be responsible for the high level of the RT activity associated with the D. simulans VLPs.

Amino Acid Sequence↗

Sensory mother cell division is specifically affected in a Cyclin-A mutant of Drosophila melanogaster.

Cyclin proteins are one of the important components of the mechanism regulating mitosis in eukaryotic cells. We isolated a Drosophila Cyclin-A mutant in which the progenitor cells of the peripheral nervous system (the sensory mother cells) do not divide properly, causing the loss and other abnormalities of mechanosensory organs in the adult fly. Sequence analysis of the mutant genome reveals that a P element is inserted into the first intron of the Cyclin-A gene. A 13 kb wild-type genomic DNA containing the Cyclin-A transcription units rescued the mutant phenotype when introduced into the mutant fly. The regulation of cell type specific expression of the Cyclin-A gene is discussed.

Animals↗

Insertional mutagenesis in Drosophila. II. P element mediated transformation of Drosophila yakuba.

Drosophila yakuba, a member of melanogaster subgroup being free of P element, acquired resistance to an antibiotic neomycin by the transformation utilizing P element. In this species, the transformation frequency was comparable to that of D. melanogaster. Further, the occurrence of 8 base pairs duplication upon the insertion of the element was confirmed. These facts suggest that the P element could be inserted into the genome in the same manner, even in D. yakuba. Any consensus for preferential insertion could not be found on the nucleotide sequence as in D. melanogaster. However, it is noticeable that a series of the short palindromic stretches was common around the insertion sites in both species. It suggests that a structural feature of DNA plays a role as a landmark for P element insertion.

Animals↗

Autoprocessing of Drosophila copia gag precursor to generate a unique laminate structure in Escherichia coli.

Drosophila copia protease is likely to be encoded in the gag gene. We have expressed copia gag polyprotein precursor in E. coli. The gag precursor was correctly processed to generate a unique laminate structure in E. coli. The processing was almost completely blocked by a mutation at the putative active site of copia protease, and resulted in accumulation of the precursor. Furthermore, the laminate structure was not found in E. coli expressing the mutant precursor. These results indicate that the protease is involved in cleaving the gag precursor itself. Also, the assembly of copia gag protein should correlate to the autoprocessing of copia gag polyprotein precursor.

Animals↗

Virus-like particle formation of Drosophila copia through autocatalytic processing.

Northern blot and nucleotide sequence analyses of copia RNA from a transfectant made by introducing a genomic copia into copia-free cells showed that the 2 kb RNA, one of the major transcripts of copia, is generated through splicing. Using the polymerase chain reaction (PCR), we have also found that the position of the splice sites used in Drosophila larvae and cultured cells originally containing copia is the same as that used in the transfectant. To investigate the function of the 2 kb RNA, we constructed mutant copias which harboured a single point mutation at the splice site or approximately 3 kb deletion of the internal region corresponding to the spliced out sequence. Analyses of transfectants made by introducing these mutant copias into copia-free cells demonstrated that the spliced 2 kb RNA contains sufficient information to make copia virus-like particles (VLPs). Furthermore, when copia RNA corresponding to the spliced RNA was translated in vitro, the major VLP protein was found to be released autocatalytically from its own precursor. A single amino acid substitution at the putative protease active site in the precursor prevented the processing, and resulted in accumulation of the mutant precursor in vitro. From these results, we conclude that copia VLPs are produced through autocatalytic processing of the precursor polyprotein encoded by the spliced copia RNA.

Amino Acid Sequence↗

Expression of inhibin alpha-subunit gene during mouse gametogenesis.

Mammalian gametogenesis is regulated through complex interactions between germ and somatic cells. To investigate the mechanism underlying the differentiation of functional gametes, some genes specifically expressed during gametogenesis have been isolated and characterized. In a search for further examples of such genes, we have isolated from a newborn mouse testis cDNA library, a clone corresponding to mouse inhibin alpha-subunit. Although it is known that the inhibin alpha-subunit molecule is abundantly produced in ovarian follicle and in testicular Sertoli cells, the spatial and temporal patterns of expression of this gene remain to be elucidated. In this study, the patterns of expression of inhibin alpha-subunit mRNA during mouse gametogenesis were examined by RNA blot, cytoplasmic dot and in situ hybridization techniques. In the testis, the concentration of inhibin alpha-subunit mRNA increased from about 16 dpc (days post coitum), peaked at birth and then gradually decreased, paralleling testicular development. Inhibin alpha-subunit mRNA was localized in Sertoli cells of wild type as well as W/Wv testes. In adult testis, mRNA was restricted to the perinuclear cytoplasm of Sertoli cells. Inhibin alpha-subunit mRNA was expressed in follicle cells of adult ovary more abundantly than in adult testis. Analysis of expression during folliculogenesis showed that the accumulation of this mRNA began in preantrum follicles and the level of expression reached a maximum in Graafian follicles.

Amino Acid Sequence↗

[A study on a design guide for detached houses with home-elevators for aged and physically handicapped persons].

The purpose of this paper is to extract the design guide of the detached houses with home-elevator for aged and physically handicapped persons, so, for 4 cases we investigated the composition of living spaces, living activities and dweller's evaluation. The main results are follows: (1) The aged and physically handicapped persons' room generally must be located at the 1st floor, but setting up the home-elevator house planning turns into more free. (2) Setting up the home-elevator, we can locate the aged and physically handicapped persons' room at the highest floor. In the case of town houses having few site areas, the highest floor has often good condition of the exposure to the sun and the view, so the aged and physically handicapped persons prefer the highest floor room. (3) In the aged and physically handicapped persons' houses, the corridor and the sanitary space get cold in winter, because those spaces are wide using the wheel chair and helping. So these houses need the floor heating. (4) In the aged and physically handicapped persons' houses, at the garage going in and out must be free without the difference in level, the space of the elevator hall must be secured fully turning the wheel chair, and at the garage the place of the outdoor wheel chair and the transfer space must be secured fully. (5) In the case of using home-elevator the noise is no problem. In opposition, the family members feel easy to hear the sound of home-elevator, because they can perceive the aged and physically handicapped persons' activities.

Aged↗

[Watery diarrhea, hypokalemia, achlorhydria syndrome due to recurrent malignant pheochromocytoma].

A case of malignant pheochromocytoma, with a recurrence 15 years after adrenalectomy and with an associated watery diarrhea, hypokalemia, achlorhydria syndrome, is reported. Histological evaluation of the tumors revealed composite malignant pheochromocytoma-ganglioneuroblastoma (well differentiated type). Vasoactive intestinal polypeptide and catecholamine levels were high both in the plasma and in the tumors. Somatostatin was also rich in the metastatic tumor of the liver, but not in the plasma. Immunohistochemical studies have demonstrated that immunoreactive vasoactive intestinal polypeptide is present in the ganglioneuroblastoma component, and that immunoreactive somatostatin is present in the pheochromocytoma component. Literature on the watery diarrhea, hypokalemia, achlorhydria syndrome associated with pheochromocytoma was reviewed.

Achlorhydria↗

Effects of methamphetamine administration on brain neurotensin-like immunoreactivity in rats.

The effects of methamphetamine (MAP) administration on neurotensin-like immunoreactivity (NTLI) were investigated in the rat brain. A single administration of MAP induced dose-dependent and reversible increases in NTLI in the caudate-putamen, limbic forebrain and midbrain. NTLI in the caudate-putamen, limbic forebrain and midbrain was also enhanced after repeated administration of MAP. It was assumed that MAP enhanced NTLI in the caudate-putamem, limbic forebrain and midbrain through its action upon the nigrostriatal and mesolimbic dopamine systems.

Animals↗

Functions of maternal mRNA as a cytoplasmic factor responsible for pole cell formation in Drosophila embryos.

Injection of mRNA extracted from Drosophila cleavage embryos or mature oocytes restored pole cell-forming ability to embryos that had been deprived of this ability by uv irradiation. However, mRNA extracted from blastoderms did not show the restoration activity. Pole cells thus formed in uv-irradiated embryos bear similarities to normal pole cells both in their morphology and their ability to migrate to the gonadal rudiments. But this mRNA does not appear to be capable of rescuing uv-induced sterility, or inducing pole cells in the anterior polar region.

Animals↗

[An autopsy case of lung cancer in a young adult dwarf].

A 31-year-old man with peripheral squamous cell carcinoma is reported. By nature he was very small. On admission he was 140 cm tall, weighed 28 kg, and was complaining of chest pain of five years duration. A giant tumor in the right lung was found to be moderately differentiated squamous cell carcinoma. He also had congenital anomalies, such as crossed ectopic kidney with fusion, café-au-lait spots, inguinal hernia, simian line, etc. He had never been exposed to cancer-causing agents. In conclusion, genetic factors related to intrauterine growth retardation might have been linked to his oncodevelopmental mechanisms.

Adult↗