Constitutive nitric oxide production in bovine aortic and brain microvascular endothelial cells: A comparative study

Chiwaka Kimura, Masahiro Oike, Keizo Ohnaka, Yoshiaki Nose, Yushi Ito

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Abstract

Vascular endothelium constitutively generates nitric oxide (NO) in large vessels and induces a relaxation of smooth muscle cells. However, little is known about the production of NO in microvessels, where smooth muscle layers are thin or absent. In this study, we have compared the constitutive production of NO in bovine brain microvascular endothelial cells (BBECs) with that in bovine aortic endothelial cells (BAECs). ATP, acetylcholine (ACh) and A23187 induced Ca2+ transients both in BBECs and BAECs. In contrast, although ATP and A23187 evoked a similar degree of [Ca2+]i increase in both types of cell, they failed to induce NO production in BBECs, as measured with an NO-sensitive fluorescent dye DAF-2, whereas in BAECs there was an increase in DAF-2 fluorescence. Hypotonic stress induced ATP release and subsequent NO production in BAECs, but not in BBECs. We have developed an in vitro model vessel system that consists of aortic smooth muscle cells embedded in a collagen gel lattice and overlaid with endothelial cells. Precontracted gels showed relaxation in response to ACh, when BAECs were overlaid. However, ACh-induced relaxation was not observed in BBEC-overlaid gels. Expression of eNOS protein as well as cellular uptake of L-[3H] arginine were significantly lower in BBECs than in BAECs. These results indicate that Ca2+-dependent NO production is at an undetectable level in BBEC, for which at least two factors, i.e. low levels of eNOS expression and L-arginine uptake, are responsible.

Original languageEnglish
Pages (from-to)721-730
Number of pages10
JournalJournal of Physiology
Volume554
Issue number3
DOIs
Publication statusPublished - Feb 1 2004

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Nitric Oxide
Endothelial Cells
Brain
Acetylcholine
Adenosine Triphosphate
Gels
Calcimycin
Smooth Muscle Myocytes
Arginine
Osmotic Pressure
Vascular Endothelium
Microvessels
Fluorescent Dyes
Smooth Muscle
Collagen
Fluorescence

All Science Journal Classification (ASJC) codes

  • Physiology

Cite this

Constitutive nitric oxide production in bovine aortic and brain microvascular endothelial cells : A comparative study. / Kimura, Chiwaka; Oike, Masahiro; Ohnaka, Keizo; Nose, Yoshiaki; Ito, Yushi.

In: Journal of Physiology, Vol. 554, No. 3, 01.02.2004, p. 721-730.

Research output: Contribution to journalArticle

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