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Pflügers Archiv - European Journal of Physiology

, Volume 455, Issue 5, pp 811–818 | Cite as

Endothelial function in aorta segments of apolipoprotein E-deficient mice before development of atherosclerotic lesions

  • Paul FransenEmail author
  • Tim Van Assche
  • Pieter-Jan Guns
  • Cor E. Van Hove
  • Gilles W. De Keulenaer
  • Arnold G. Herman
  • Hidde Bult
Cardiovascular System

Abstract

Acetylcholine (ACh)-induced relaxation declines in apolipoprotein E-deficient (apoE−/−) mouse aortas, but only after atherosclerotic plaque formation. This study investigated intracellular calcium concentrations [Ca2+]i and changes in phenylephrine-induced contractions as index of baseline nitric oxide (NO) bioavailability before plaque development. Isometric contractions of thoracic aorta rings of young (4 months) apoE−/− and C57BL/6J (WT) mice were evoked by phenylephrine (3 × 10−9–3 × 10−5 M) in the presence and absence of endothelial cells (ECs) or NO synthase (NOS) inhibitors. [Ca2+]i (Fura-2 AM) and endothelium-dependent relaxation were measured at baseline and after ACh stimulation. Segments of apoE−/− mice were significantly more sensitive and developed more tension than WT segments in response to phenylephrine. The differences disappeared after NOS inhibition or EC removal or upon increasing [Ca2+]i in apoE−/− strips with 10−6 M cyclopiazonic acid or 10−7 M Ca2+-ionophore A23187. Expression of endothelial NOS (eNOS) mRNA was similar in apoE−/− and WT aorta segments. Basal [Ca2+]i was significantly lower in apoE−/− than in WT strips. Relaxation by ACh (3 × 10−9–10−5 M) was time- and dose-dependently related to [Ca2+]i, but neither ACh-induced relaxation nor Ca2+ mobilization were diminished in apoE−/− strips. In conclusion, basal, but not ACh-induced NO bioavailability, was compromised in lesion-free aorta of apoE−/− mice. Decreased basal NO bioavailability was not related to lower eNOS expression, but most likely related to lower basal [Ca2+]i. These findings further point to important differences between basal and stimulated eNOS activity.

Keywords

Atherosclerosis Apolipoprotein E-deficient mouse Endothelial cell Intracellular calcium [Ca2+]i Nitric oxide synthase Thoracic aorta Basal nitric oxide 

Abbreviations

ACh

acetylcholine

apoE−/−

apolipoprotein E-deficient

[Ca2+]i

intracellular calcium concentration

CPA

cyclopiazonic acid

EC

endothelial cell

l-NAME

\( N^{\omega } \)-nitro-l-arginine methyl ester

l-NNA

\( N^{\omega } \)-nitro-l-arginine

NO

nitric oxide

NOS

nitric oxide synthase

eNOS

endothelial nitric oxide synthase

WT

wild type

Notes

Acknowledgments

The technical assistance of Valerie Croons and Ludo Zonnekeyn and the secretarial help of Liliane Van den Eynde are greatly appreciated. This work was supported by grants from Bijzonder Onderzoek Fonds of the University of Antwerp to Paul Fransen (project 1083, BOF KP 2005) and Fonds voor Wetenschappelijk Onderzoek (FWO, Vlaamse Gemeenschap, project G.0174.06).

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Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Paul Fransen
    • 1
    Email author
  • Tim Van Assche
    • 1
  • Pieter-Jan Guns
    • 1
  • Cor E. Van Hove
    • 1
  • Gilles W. De Keulenaer
    • 1
  • Arnold G. Herman
    • 1
  • Hidde Bult
    • 1
  1. 1.Division of Pharmacology, Faculties of Medicine and Pharmaceutical SciencesUniversity of AntwerpWilrijkBelgium

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