Myoendothelial gap junctions may provide the pathway for EDHF in mouse mesenteric artery.
Endothelium-dependent hyperpolarization of vascular smooth muscle provides a major pathway for relaxation in resistance arteries. This can occur due to direct electrical coupling via myoendothelial gap junctions (MEGJs) and/or the release of factors (EDHF). Here we provide evidence for the existence of functional MEGJs in the same, defined branches of BALB/C mouse mesenteric arteries which show robust EDHF-mediated smooth muscle relaxation. Cyclopiazonic acid (CPA, 10 microM) was used to stimulate EDHF in arteries mounted under isometric conditions and constricted with phenylephrine. Simultaneous measurement of smooth muscle membrane potential and tension demonstrated that CPA caused a hyperpolarization of around 10 mV, reversing the depolarization to phenylephrine by 94% and the associated constriction by 66%. The relaxation to CPA was endothelium dependent, associated with the opening of Ca2+-activated K channels, and only in part due to the release of nitric oxide (NO). In the presence of the NO synthase inhibitor, L-NAME (100 microM), the relaxation to CPA could be almost completely inhibited with the putative gap junction uncoupler, carbenoxolone (100 microM). Inhibition of the synthesis of prostaglandins or metabolites of arachidonic acid had no effect under the same conditions, and small rises in exogenous K+ failed to evoke consistent or marked smooth muscle relaxation, arguing against a role for these molecules and ions as EDHF. Serial section electron microscopy revealed a high incidence of MEGJs, which was correlated with heterocellular dye coupling. Taken together, these functional and morphological data from a defined mouse resistance artery suggest that the EDHF response in this vessel may be explained by extensive heterocellular coupling through MEGJs, enabling spread of hyperpolarizing current.
Department of Pharmacy and Pharmacology, University of Bath, Bath, UK. k.a.dora@bath.ac.uk
Kim A
KA
Dora
Shaun L
SL
Sandow
Nicola T
NT
Gallagher
Hiromichi
H
Takano
Nicole M
NM
Rummery
Caryl E
CE
Hill
Chris J
CJ
Garland
D000109:Acetylcholine / Q000494:pharmacology; D000818:Animals; D001685:Biological Factors / Q000502:physiology*; D004730:Endothelium, Vascular / Q000502:physiology*; D017629:Gap Junctions / Q000502:physiology*; D007211:Indoles / Q000494:pharmacology; D008297:Male; D008564:Membrane Potentials; D008638:Mesenteric Arteries / Q000502:physiology* / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D011188:Potassium / Q000494:pharmacology
D001685:Biological Factors; D007211:Indoles; C067410:endothelium-dependent hyperpolarization factor; D000109:Acetylcholine; D011188:Potassium; C000543:cyclopiazonic acid