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Yoichi Matsuda

Publications and source records attributed to Yoichi Matsuda.

46 records · Page 3Linked to original sources

Phenotypic analysis of Meltrin alpha (ADAM12)-deficient mice: involvement of Meltrin alpha in adipogenesis and myogenesis.

Meltrin alpha (ADAM12) is a metalloprotease-disintegrin whose specific expression patterns during development suggest that it is involved in myogenesis and the development of other organs. To determine the roles Meltrin alpha plays in vivo, we generated Meltrin alpha-deficient mice by gene targeting. Although the number of homozygous embryos are close to the expected Mendelian ratio at embryonic days 17 to 18, ca. 30% of the null pups born die before weaning, mostly within 1 week of birth. The viable homozygous mutants appear normal and are fertile. Most of the muscles in the homozygous mutants appear normal, and regeneration in experimentally damaged skeletal muscle is unimpeded. In some Meltrin alpha-deficient pups, the interscapular brown adipose tissue is reduced, although the penetrance of this phenotype is low. Impaired formation of the neck and interscapular muscles is also seen in some homozygotes. These observations suggest Meltrin alpha may be involved in regulating adipogenesis and myogenesis through a linked developmental pathway. Heparin-binding epidermal growth factor-like growth factor (HB-EGF) is a candidate substrate of Meltrin alpha, and we found that TPA (12-O-tetradecanoylphorbol-13-acetate)-induced ectodomain shedding of HB-EGF is markedly reduced in embryonic fibroblasts prepared from Meltrin alpha-deficient mice. We also report here the chromosomal locations of Meltrin alpha in the mouse and rat.

ADAM Proteins↗

Characterization of Sleeping Beauty transposition and its application to genetic screening in mice.

The use of mutant mice plays a pivotal role in determining the function of genes, and the recently reported germ line transposition of the Sleeping Beauty (SB) transposon would provide a novel system to facilitate this approach. In this study, we characterized SB transposition in the mouse germ line and assessed its potential for generating mutant mice. Transposition sites not only were clustered within 3 Mb near the donor site but also were widely distributed outside this cluster, indicating that the SB transposon can be utilized for both region-specific and genome-wide mutagenesis. The complexity of transposition sites in the germ line was high enough for large-scale generation of mutant mice. Based on these initial results, we conducted germ line mutagenesis by using a gene trap scheme, and the use of a green fluorescent protein reporter made it possible to select for mutant mice rapidly and noninvasively. Interestingly, mice with mutations in the same gene, each with a different insertion site, were obtained by local transposition events, demonstrating the feasibility of the SB transposon system for region-specific mutagenesis. Our results indicate that the SB transposon system has unique features that complement other mutagenesis approaches.

Animals↗

[Spinal anesthesia using hyperbaric bupivacaine HCl for cesarean section].

BACKGROUND: We performed this prospective study to determine the proper amount of hyperbaric bupivacaine hydrochloride as a spinal anesthetic agent for cesarean section. METHODS: The parturients were randomly allocated to receive one of four spinal agents in a blind manner; tetracaine 10 mg (control), bupivacaine 10, 12.5 and 15 mg. Morphine HCl 0.1 mg was added to each agent and the total volume was adjusted to 3.1 ml with 10% glucose solution. RESULTS: All the four spinal agents provided an adequate analgesic level (T 5) without serious complications. Among the three dosages of bupivacaine, the time interval requiring for anesthetic level to reach T 5 tended to be shorter with a larger amount of bupivacaine. The incidence of intraoperative supplemental analgesic and hypotension and the dosage of ephedrine used to treat hypotension were greater in the patients anesthetized with tetracaine 10 mg than in those anesthetized with bupivacaine 10 mg, which is equipotent to tetracaine 10 mg. CONCLUSIONS: 1. As a spinal anesthetic agent for cesarean section, hyperbaric bupivacaine is superior to tetracaine. 2. Hyperbaric bupivacaine 10 mg, 12.5 mg or 15 mg can be used safely and effectively as a spinal agent for cesarean section. 3. High dose bupivacaine is recommended in an urgent case, and low dose bupivacaine is recommended when maternal hypotension must be strictly avoided.

Anesthesia, Obstetrical↗

Molecular and cytogenetic characterization of the mouse ATP-binding cassette transporter Abcg4.

We have cloned a new mouse ATP-binding cassette (ABC) transporter, Abcg4, from a complementary DNA (cDNA) library of mouse brain. The cloned Abcg4 cDNA encodes a protein consisting of 646 amino acids and including one ATP-binding cassette and six transmembrane domains. The Abcg4 protein exhibits high identity (96%) with human ABCG4 in terms of the amino acid sequence. Fluorescence in situ hybridization with mouse and rat chromosomes has revealed that the Abcg4 gene is located on chromosomes 9A5.3 and 8q22 distal in mouse and rat, respectively. In these loci on mouse and rat chromosomes, conserved linkage homologies were hitherto identified with human chromosome 11q23, which involves the human ABCG4 gene. The mouse Abcg4 gene as well as the human ABCG4 gene each has a total of 14 exons to encode its respective protein. High transcript levels of mouse Abcg4 were detected in mouse brain, spleen, eye, and bone marrow. Taken together, our data on the chromosomal location, gene homology, protein structure, and phylogenetic relationships strongly support the idea that mouse Abcg4 is orthologue to the human ABCG4. By functionally analyzing the mouse Abcg4 protein, we may better understand the biological role of the human ABCG4 transporter.

ATP Binding Cassette Transporter, Subfamily G↗

The ADAM1a and ADAM1b genes, instead of the ADAM1 (fertilin alpha) gene, are localized on mouse chromosome 5.

Fertilin is reported to be a heterodimeric protein composed of A Disintegrin And Metalloprotease 1 (ADAM1, fertilin alpha) and ADAM2 (fertilin beta) located on the sperm surface. In the process of clarifying the molecular basis of mouse ADAM1, we have identified two intron-less mouse genes encoding different isoforms of ADAM1, termed ADAM1a and ADAM1b. The amino acid sequences of ADAM1a and ADAM1b deduced from the DNA sequences were homologous to each other (99% identity) in the pro- and metalloprotease domains, whereas the C-terminal half region of ADAM1a, including the disintegrin and Cys-rich domains, shared only a low degree of identity (37%) with that of ADAM1b. These two genes were both localized on mouse chromosome 5 as a single copy gene, and were expressed specifically in the testis. These data demonstrate the presence of the ADAM1a (Adam1a) and ADAM1b (Adam1b) genes in mouse, instead of the ADAM1 gene, and may imply different roles of ADAM1a and ADAM1b in spermatogenesis, sperm maturation, and/or fertilization.

ADAM Proteins↗

FISH analysis of 142 EGFP transgene integration sites into the mouse genome.

Production of transgenic animals is an important technique for studying various biological processes. However, whether the integration of a particular transgene occurs randomly in the mouse genome has not been determined. Analysis by fluorescence in situ hybridization of the integration sites of the 142 EGFP (a mutant of green fluorescent protein) transgenic lines that we produced showed that the transgenes had become incorporated into every mouse chromosome. A single integration site was observed in 82.4% of the lines. The concomitant integrations of transgene into two different loci were observed in 15 cases (10.6%). In 3 cases, the transgenic founder mice showed chimerism in integration sites (2.1%). Chromosomal translocation was observed in 7 cases (4.9%). Moreover, when we statistically analyzed the transgene integration sites of these mouse lines, they were shown to distribute unevenly throughout the genome. This is the first report to analyze the transgene integration sites by producing more than 100 transgenic mouse lines.

Animals↗

Characterization and chromosomal distribution of novel satellite DNA sequences of the lesser rhea (Pterocnemia pennata) and the greater rhea (Rhea americana).

Two different types of novel satellite DNA (stDNA) sequences were cloned from the lesser rhea (Ptercnemia pennata) and the greater rhea (Rhea americana) after digestion of genomic DNAs with a restriction endonuclease Pvu II, and characterized by filter hybridization and in-situ hybridization to metaphase chromosomes. These nucleotide sequences consisted of GC-rich 288-bp and 332-bp repeated elements in P. pennata and 288-bp and 336-bp repeated elements in R. americana, all of which were organized in tandem arrays in the genome. The 288-bp and 332-bp elements of P. pennata displayed strong sequence similarity with the 288-bp and 336-bp elements of R. americana, respectively. The 332-bp and 336-bp elements were located on almost all the microchromosomes in both the species. The other type of repeated elements, the 288-bp element, was located on four and nine pairs of microchromosomes in P. pennata and R. americana, respectively. All the stDNA sequences were not crosshybridized to genomic DNAs of another three ratite species, ostrich (Struthio camelus), cassowary (Casuarius casuarius) and emu (Dromaius novaehollandiae), suggesting that these stDNA sequences are conserved in the same family but fairly divergent among the different families of Struthioniformes.

Animals↗

Identification and expression of mouse Langerin (CD207) in dendritic cells.

We have cloned the mouse homologue of human Langerin (h-Langerin), a type II transmembrane protein with a single external C-type lectin domain. Mouse Langerin (m-Langerin) displays 65 and 74% homologies in total amino acid and lectin domains with those of h-Langerin. The cognate mouse and rat genes were assigned to chromosome 6D1-D2 and chromosome 4q33 distal-q34.1 proximal respectively, syntenic to the h-Langerin gene on chromosome 2p13. With RT-PCR, m-Langerin transcripts were as expected detected in MHC class II+, but not MHC class II-, cells from epidermis and the expression level was reduced by culture. However, m-Langerin transcripts were also expressed in spleen, lymph nodes (LN), thymus, liver, lung and even heart, but not gut-associated lymphoid tissues. In single-cell lymphoid suspensions, m-Langerin transcripts were mainly detected in the CD11c+ dendritic cells (DC), especially the CD11blow/CD8high fraction of spleen and LN. DC generated from bone marrow precursors by granulocyte macrophage colony stimulating factor (GM-CSF) expressed m-Langerin, but this was shut down during maturation with CD40 ligand or lipopolysaccharide. DC derived from blood monocytes by GM-CSF + IL-4 lacked m-Langerin unless the cultures were supplemented with transforming growth factor (TGF)-beta1. Unexpectedly, significant amounts of m-Langerin transcripts were detected in skin and LN of TGF-beta1-deficient mice, although in much lower amounts than littermate controls. Recombinant m-Langerin could form multimers and bind to mannan-agarose. These findings indicate that Langerin expression is regulated at several levels: by TGF-beta1, DC subsets, DC maturation and the tissue environment.

Amino Acid Sequence↗

b-series Ganglioside deficiency exhibits no definite changes in the neurogenesis and the sensitivity to Fas-mediated apoptosis but impairs regeneration of the lesioned hypoglossal nerve.

The polymorphic carbohydrate structures of gangliosides play regulatory roles. In particular, b-series gangliosides, all of which contain alpha-2,8 sialic acids, have been considered to be critical in various biological events such as adhesion, toxin binding, neurite extension, cell growth, and apoptosis. To clarify the physiological functions of b-series gangliosides in vivo, we have established a gene knockout mouse of GD3 synthase. Although all b-series structures were deleted in the mutant mice, they showed an almost complete nervous tissue morphology with no apparent abnormal behavior. Moreover, no differences in Fas-mediated apoptotic reaction of lymphocytes between wild type and the mutant mice were detected. However, the mutant mice exhibited clearly reduced regeneration of axotomized hypoglossal nerves compared with the wild type, suggesting that b-series gangliosides are more important in the repair rather than in the differentiation of the nervous system and apoptotic process induced via Fas.

Animals↗