Muscarinic receptors and calcium signalling in rabbit isolated pulmonary artery smooth muscle cells

G.T. O'Neill, E.G. Rowan, A.M. Gurney

Research output: Contribution to journalArticle

Abstract

Acetylcholine (ACh) is an endothelium-dependent vasodilator, but when pulmonary artery endothelium is damaged it causes vasoconstriction through a direct action on smooth muscle muscarinic receptors (Dinh-Xuan et al., 1989). In rabbit main pulmonary artery this action involves M2 and M3 receptors The muscarinic agonist, oxotremorine sesquifumarate (Oxo-S), also contracts this preparation, but the effects appear to be mediated solely through M2 receptors This study characterised responses to ACh and Oxo-S in isolated pulmonary artery smooth muscle cells (PASMC) by monitoring changes in the fluorescence of the Ca2+ indicator, fluo-4. Arteries were obtained from male New Zealand rabbits (2-2.5 kg) killed by lethal injection of sodium pentobarbitone (140 mg kg-' i.v.). PASMC were isolated by enzymatic digestion, loaded with 1 jM fluo-4 at 221C for 45 min, washed and bathed in physiological salt solution (PSS). Fluorescence was excited at 488 nm and measured between 503 and 553 nm using a confocal microscope. Calcium signals were quantified as the change in fluorescence (AF) relative to the background fluorescence (F0). Ca2+-free solution was prepared by replacing CaCl2 in the PSS with equimolar MgC12 and adding 5mM EGTA. To study Gi involvement, PASMC were incubated with pertussis toxin (PTX; 5gig ml-') for 3-4 hr at 22°C and compared with control cells treated in the same way, but without exposure to PTX. Experiments were performed 22°C. Results are expressed as mean ± s.e.m.
LanguageEnglish
Pages24P-24P
JournalBritish Journal of Pharmacology
Volume137
Issue numberSuppl.
Publication statusPublished - Dec 2002

Fingerprint

Calcium Signaling
Muscarinic Receptors
Pulmonary Artery
Smooth Muscle Myocytes
Rabbits
Fluorescence
Arteries
Capital Punishment
Endothelium-Dependent Relaxing Factors
Muscarinic Agonists
Egtazic Acid
Pertussis Toxin
Pentobarbital
Acetylcholine
Endothelium
Digestion
Calcium
Fluo 4

Keywords

  • muscarinic receptors
  • calcium signalling
  • pulmonary artery smooth muscle cells

Cite this

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abstract = "Acetylcholine (ACh) is an endothelium-dependent vasodilator, but when pulmonary artery endothelium is damaged it causes vasoconstriction through a direct action on smooth muscle muscarinic receptors (Dinh-Xuan et al., 1989). In rabbit main pulmonary artery this action involves M2 and M3 receptors The muscarinic agonist, oxotremorine sesquifumarate (Oxo-S), also contracts this preparation, but the effects appear to be mediated solely through M2 receptors This study characterised responses to ACh and Oxo-S in isolated pulmonary artery smooth muscle cells (PASMC) by monitoring changes in the fluorescence of the Ca2+ indicator, fluo-4. Arteries were obtained from male New Zealand rabbits (2-2.5 kg) killed by lethal injection of sodium pentobarbitone (140 mg kg-' i.v.). PASMC were isolated by enzymatic digestion, loaded with 1 jM fluo-4 at 221C for 45 min, washed and bathed in physiological salt solution (PSS). Fluorescence was excited at 488 nm and measured between 503 and 553 nm using a confocal microscope. Calcium signals were quantified as the change in fluorescence (AF) relative to the background fluorescence (F0). Ca2+-free solution was prepared by replacing CaCl2 in the PSS with equimolar MgC12 and adding 5mM EGTA. To study Gi involvement, PASMC were incubated with pertussis toxin (PTX; 5gig ml-') for 3-4 hr at 22°C and compared with control cells treated in the same way, but without exposure to PTX. Experiments were performed 22°C. Results are expressed as mean ± s.e.m.",
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Muscarinic receptors and calcium signalling in rabbit isolated pulmonary artery smooth muscle cells. / O'Neill, G.T.; Rowan, E.G.; Gurney, A.M.

In: British Journal of Pharmacology, Vol. 137, No. Suppl., 12.2002, p. 24P-24P.

Research output: Contribution to journalArticle

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AU - Gurney, A.M.

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