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J Physiol. 2004 Mar 1;555(Pt 2):459-69. Epub 2003 Dec 23.

Increase in endothelial cell Ca(2+) in response to mouse cremaster muscle contraction.

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  • 1Department Pharmacology and Physiology, University of Rochester Medical Center, Box 711, Rochester, NY 14642, USA.


We addressed the role of endothelial cells (ECs) in metabolic dilatation of skeletal muscle arterioles in anaesthetized mice in situ. Electrical field stimulation was used to contract the cremaster muscle for 15 s at 30 Hz. Diameter was observed using bright field microscopy. In controls, muscle contraction produced a 15.7 +/- 1.5 microm dilatation from a baseline of 17.4 +/- 1.6 microm. Endothelial denudation (-EC) via intraluminal perfusion of air abolished this response (1.6 +/- 1.2 microm in -EC, P < 0.05), identifying endothelium as the primary vascular cell type initiating the dilatation. To investigate the role of EC Ca(2+) in metabolic dilatation, arteriolar ECs were loaded with Fluo-4 AM or BAPTA AM by intraluminal perfusion, after which blood flow was re-established. Ca(2+) activity of individual ECs was monitored as a function of change from baseline fluorescence using confocal microscopy. In ECs, whole cell Ca(2+) increased (>10%, P < 0.05) during muscle contraction, and localized Ca(2+) transients were increased (>20%, P < 0.05) during the first minute after contraction. Chelation of EC Ca(2+) abolished the dilatations in response to muscle contraction (1.1 +/- 0.7 microm, P < 0.05). Inhibition of P(1) purinergic receptors (with xanthine amine congener) did not alter the rate of onset of the dilatation (P > 0.05) but decreased its magnitude immediately post stimulation (7.1 +/- 0.9 microm, P < 0.05) and during recovery. These findings demonstrate obligatory roles for endothelium and EC Ca(2+) during metabolic dilatation in intact arterioles. Furthermore, they suggest that at least two separate pathways mediate the local response, one of which involves stimulation of endothelial P(1) purinergic receptors via endogenous adenosine produced during muscle activity.

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