アンチセンスオリゴDNAを用いた細胞膜カルシウムポンプ活性の選択的抑制

DOI 参考文献11件

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タイトル別名
  • Selective blockade of plasma membrane Ca<SUP>2+</SUP> pump activity with antisense oligodeoxynucleotides

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The importance of the plasma membrane Ca2+-pump in regulating the intracellular Ca2+ concentrations ([Ca2+]i) has been recognized. However, the exact role of the plasma membrane Ca2+ pump in the intracellular Ca2+ metabolism of living cells remains to be elucidated. To answer this query antisense oligodeoxynucleotides based on the sequence of the rat plasma membrane Ca2+ pump (PMCA-1) were used to study the effects of inhibiting expression of this protein on the regulation of [Ca2+]i in cultured vascular smooth muscle cells. Antisense oligodeoxynucleotides targeting a sequence [position +1 to +18] of rat PMCA-1 reduced the expression of the pump in a concentration dependent manner, while mismatch or sense oligodeoxynucleotides had no effects. Cytotoxic effects of antisense oligonucleotides were not observed. Antisense treatment affected not only the resting level of [Ca2+]i but also the magnitude of the increase in [Ca2+]i of vascular smooth muscle cells induced by Ca2+ mobilizing agents. 45Ca efflux from vascular smooth muscle cells mediated by plasma membrane Ca2+ pump was also inhibited by the treatment with antisense oligodeoxynucleotides. These results suggest that reduced expression of the plasma membrane Ca2+ pump (PMCA-1) significantly alters intracellular Ca2+ regulation of vascular smooth muscle cells and that PMCA-1 plays important roles in the regulation of intracellular Ca2+ concentrations of vascular smooth muscle cells both in the resting and stimulated states. Antisense oligodeoxynucleotides targeting a sequence of the pump becomes a powerful tool to investigate its physiological role.

収録刊行物

  • 日本薬理学雑誌

    日本薬理学雑誌 108 (supplement), 99-103, 1996

    公益社団法人 日本薬理学会

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