Acute effects of cell isolation on InsP profiles in adult rat cardiomyocytes

Journal of Molecular and Cellular Cardiology
E A Woodcock, K A Lambert

Abstract

The isolation and culture of adult rat cardiomyocytes was shown to cause major changes in the contents of [3H]-labeled inositol phosphates and inositol phospholipids. Undigested heart tissue contained high levels of [3H]Ins(1,4,5)P3 (5364+/-800 ct/min/g tissue, 80+/-12 ct/min/mg protein) and mass content averaged 13.8 nmol/g tissue or 208+/-36 pmol/mg protein (mean+/-S.E.M., n=4). After collagenase digestion, [3H]Ins(1,4,5)P3 was undetectable and the mass content of Ins(1,4,5)P3 had decreased to 0.8+/-0.2 pmol/mg protein (mean+/-S.E.M., n=4, P<0.01). [3H]Ins(1,4)P2 was reduced by 80% and [3H]PtdIns(4,5)P2 by 90%. These profiles remained essentially unchanged when the isolated cells were maintained in culture for up to 24 h, even though the inositol phosphate response remained sensitive to norepinephrine. Similar to findings in intact tissue, the inositol phosphate response to norepinephrine in these cells was inhibited by neither U-73122 (5 microM) nor by neomycin (5 mM). By 48 h in culture, the relative levels of [3H]Ins(1,4,5)P3 and [3H]Ins(1,4)P2 had increased in relation to the total inositol phosphate content and responses appeared to better reflect intact tissue. However, while retaining insensitivity to neomycin, cells a...Continue Reading

Citations

Mar 23, 2010·American Journal of Physiology. Heart and Circulatory Physiology·Einsley JanowskiMartin Morad
Jan 5, 2007·Proceedings of the National Academy of Sciences of the United States of America·Julie R McMullenGarry L Jennings
Jul 8, 2008·Journal of Molecular and Cellular Cardiology·Jens KockskämperMartin D Bootman
Dec 6, 2001·Science's STKE : Signal Transduction Knowledge Environment·D W HilgemannC Nasuhoglu
Jun 11, 2002·American Journal of Physiology. Cell Physiology·Cem NasuhogluDonald W Hilgemann
Dec 1, 2004·Basic & Clinical Pharmacology & Toxicology·Dagny SandnesJan-Bjørn Osnes

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