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Publications (9)23.28 Total impact

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    ABSTRACT: The dedifferentiation of vascular smooth muscle cells (VSMCs) plays a critical role in the progression of atherosclerosis and restenosis after angioplasty. Thus, factors that stimulate smooth muscle cell differentiation should be useful for therapy for these diseases. Previously, we found that l-ascorbic acid (L-Asc) induces the expression of smooth muscle-specific genes in a pluripotent bone marrow stromal cell line, TBR-B. This finding suggests that l-Asc stimulates the differentiation of smooth muscle cells. In the present study, we investigated the effects of l-Asc and its derivatives on the differentiation state of VSMCs in vitro and in vivo. l-Asc and its long-lasting derivatives stimulated the production of smooth muscle-specific myosin heavy chain-1 (SM1) and calponin 1 in a dose-dependent manner in rat cultured VSMCs, and the elevated production of SM1 and calponin 1 was maintained for at least 2 weeks. Moreover, oral administration of 3 g/kg of l-Asc to the balloon-injured rats induced a higher expression of SM1 and calponin 1 in the injured arteries compared with that from administration of the delivery vehicle alone. These data demonstrated new biologic activity, such as the stimulation of VSMC differentiation, of l-Asc and its long-lasting derivatives. In addition, these compounds may serve as useful tools for analysis of the differentiation of VSMCs and for therapy for vascular diseases.
    No preview · Article · Jan 2004 · Journal of Cardiovascular Pharmacology
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    ABSTRACT: We established an in vitro culture system which mimicked the differentiation pathway of smooth muscle cell, using TBR-B, a bone marrow stromal cell line derived from transgenic mice harboring temperature-sensitive SV40 large T-antigen gene. TBR-B cells have the potential to express smooth muscle-specific genes including h1-calponin, h-caldesmon, SM22alpha and alpha-actin, only after cultured in the differentiation medium for 2 weeks. The differentiation state of TBR-B was well controlled by using different culture medium. Using this cell line, we also found that ascorbic acid is a potent factor inducing the expression of h1-calponin and alpha-actin. TBR-B cells will serve as a useful tool for elucidating the regulatory mechanisms of smooth muscle-specific gene expression, and for identifying compounds that regulate the differentiation state of vascular smooth muscle cells.
    Preview · Article · Oct 2000 · FEBS Letters
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    Tomoyasu Sugiyama · Yuzuru Matsuda · Katsuhiko Mikoshiba
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    ABSTRACT: The linkage between inositol 1,4,5-trisphosphate (IP(3)) receptors (IP(3)Rs) and cytoskeletal proteins is considered to be important in cell function. In the present study, the association of IP(3)R subtypes with cytoskeletal proteins was examined using monoclonal antibodies specific to each IP(3)R subtype. We found that IP(3)R type 2 colocalized with talin, a focal contact cytoskeletal protein. IP(3)R type 2 exhibited a patchy distribution in the peripheral cytoplasm differently from type 1 and type 3 IP(3)R. Furthermore, IP(3)R subtypes co-immunoprecipitated with talin, vinculin and alpha-actin, but not alpha-actinin or paxillin.
    Full-text · Article · Feb 2000 · FEBS Letters
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    ABSTRACT: Proliferation of smooth muscle cells (SMC) has a role in the development of cardiovascular diseases. We investigated the alteration of contractile signals in proliferating SMC by measuring the increase in intracellular [Ca(2+)] to endothelin-1 (ET-1), noradrenaline (NA), or angiotensin II (AgII). We found that the increase in intracellular [Ca(2+)] by NA or ET-1 decreased in proliferating SMC in comparison to growth-arrested SMC. The increase in intracellular [Ca(2+)] by AgII was stable between the cells. Immunoblotting of inositol 1,4,5-trisphosphate receptors (IP(3)Rs) which are responsible for the mobilization of Ca(2+) by those vasoactive substances revealed that expression of IP(3)R type 1 and type 2 was decreased. Expression of IP(3)R type 3 was increased. The altered Ca(2+) signaling by the cell growth might involve the expression of IP(3)R subtypes.
    No preview · Article · Dec 1999 · Biochemical and Biophysical Research Communications
  • Yuzuru Matsuda · Kazuhide Hasegawa
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    ABSTRACT: We have established a novel smooth muscle cell line (SVS) which retains the expression of specific markers for smooth muscle cells, such as smooth muscle myosin-1, calponin and SM22 alpha, from temperature-sensitive SV40 large T-antigen transgenic mice. SVS cells showed temperature-dependent growth and the expression of smooth muscle markers, showing that the differentiation stage can be controlled in this cell line by culture temperature. We have constructed luciferase-reporter cell lines for calponin and SM22 alpha genes by using SVS cells. We detected some compounds including ascorbic acid, which stimulated the expression of smooth muscle markers by the reporter assay. Ascorbic acid and its long lasting derivatives stimulated the expression of markers continuously. Moreover, ascorbic acid activated the expression of markers in synthetic SMCs seen in the intima of a rat balloon injury model. Thus, this reporter system should be useful for discovery of new antiatherosclerotic drugs.
    No preview · Article · Nov 1999 · Folia Pharmacologica Japonica
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    ABSTRACT: Recently we have established an aortic smooth muscle cell line, p53LMAC01 obtained from p53 knockout mice. This cell line showed some differentiated properties which were accelerated by 5-azacytidine treatment [1]. In this study, further characterization of p53LMAC01 cell line was investigated according to cell growth and differentiation, and especially focused into the changes of cell feature, actin filaments' formation, and changes of intracellular calcium concentrations to sympathetic nerve transmitter, norepinephrine. While the cell feature was changed from flattened shape to extended form during 4 days, actin filaments were developing, arranging in parallel to longitudinal direction, and gathering under the surface membrane. In 11 days many cells died and detached from substrate, while actin filaments became poor except for the surface membrane in the remained cells. Appearance of calcium response to noradrenalin needed several days after passage as well as a morphological change of the cells for the extended form and development of actin filaments. The calcium response was maintained on 11 days, which coincided with the result that the cells hold actin filaments under the surface membrane. These results suggest that p53LMAC01 cell line maintains several differentiated characters of adult smooth muscle cell and that their expression needs several days after passage.
    No preview · Article · Feb 1999 · Molecular and Cellular Biochemistry
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    ABSTRACT: The expression of the contractile proteins, alpha-actin, smooth muscle myosin heavy chain-1 (SM1) and calponin present in smooth muscle cells (SMC) in the presence of oxidized low density lipoproteins (oxLDL) was investigated in two different cell cultures: the mouse smooth muscle cell line SVSC and rat smooth muscle cells (RSMC). Exposure of the cells to 187 microg protein/ml oxLDL for 24 h reduced the expression of all three contractile proteins in both cell cultures when compared to cells incubated in the presence of native LDL. This investigation of the response of SMC contractile proteins to oxLDL may provide further insights into the mechanisms by which oxidatively modified LDL is atherogenic and suggests that oxLDL may contribute to the regulation of the expression of the genes responsible for the synthesis of smooth muscle cell contractile proteins.
    Full-text · Article · Apr 1998 · FEBS Letters
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    ABSTRACT: We have established novel vascular smooth muscle cell lines (SVS30 and SVS24 cells) which retain the expression of specific markers for smooth muscle cells, such as alpha-actin, smooth muscle myosin heavy chain-1, and calponin, from transgenic mice harboring the temperature-sensitive SV40 large T-antigen gene. SVS cell lines showed temperature-dependent growth and the expression of SV40 large T-antigen. Interestingly, protein and mRNA levels of smooth muscle myosin heavy chain-1 and calponin seen in culture at the non-permissive temperature (39 degrees C) were higher than those at the permissive temperature (33 degrees C). These results suggest that SV40 large T-antigen affects the expression of smooth muscle-specific markers in SVS cell lines, and that some of the characters in SVS cell lines can be controlled by culture temperature. SVS cell lines should be quite valuable tools with which to study the regulation of phenotypic modulation of smooth muscle cells, and to identify smooth muscle specific transcription factors which involve the expression of smooth muscle myosin heavy chain-1 and calponin genes.
    No preview · Article · Sep 1997 · Journal of Molecular and Cellular Cardiology
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    Kazuhide Hasegawa · Emi Arakawa · Shoji Oda · Yuzuru Matsuda
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    ABSTRACT: The nucleotide and deduced amino acid sequences of two isoforms of mouse smooth-muscle myosin heavy chain (SM1 and SM2) were determined. SM1 (6175 bp) and SM2 (6214 bp) cDNA contained a single open reading frame that encodes 1972 and 1938 amino acids (227,056 Da and 223,294 Da), respectively. Smooth muscle myosin heavy chain mRNA was expressed highly in smooth muscle tissue (small intestine) and weakly in heart and lung. Each of SM1 and SM2 cDNA was transfected and expressed in CHO cells. The expressed myosin heavy chains were detected with an antibody raised against smooth muscle myosin heavy chains and showed the same mobility as the native smooth muscle myosin heavy chains in SDS-PAGE.
    Preview · Article · Apr 1997 · Biochemical and Biophysical Research Communications