Electrical conduction along endothelial cell tubes from mouse feed arteries: confounding actions of glycyrrhetinic acid derivatives

• Published in: British journal of pharmacology 2012-05, Vol.166 (2), p.774-787
• Main Authors: Behringer, Erik J, Socha, Matthew J, Polo‐Parada, Luis, Segal, Steven S
• Format: Article
• Language: English
• Subjects: Abdomen; Acetylcholine - pharmacology; Animals; Arteries - drug effects; Arteries - physiology; Biological and medical sciences; calcium‐activated K+ channels; Carbenoxolone - pharmacology; cell‐to‐cell coupling; Dyes; Electric Conductivity; Endothelial Cells - drug effects; Endothelial Cells - physiology; Endothelium; gap junction channels; Gap Junctions - drug effects; Gap Junctions - physiology; Glycyrrhetinic Acid - pharmacology; hyperpolarization; Indoles - pharmacology; Intermediate-Conductance Calcium-Activated Potassium Channels - physiology; Medical sciences; Membrane Potentials; Mice; Mice, Inbred C57BL; microcirculation; Muscle, Skeletal - physiology; Oximes - pharmacology; Pharmacology. Drug treatments; Research Papers; resistance arteries; Rodents; Small-Conductance Calcium-Activated Potassium Channels - physiology
• ISSN: 0007-1188; 1476-5381
• DOI: 10.1111/j.1476-5381.2011.01814.x

BACKGROUND AND PURPOSE Electrical conduction along endothelium of resistance vessels has not been determined independently of the influence of smooth muscle, surrounding tissue or blood. Two interrelated hypotheses were tested: (i) Intercellular conduction of electrical signals is manifest in endothelial cell (EC) tubes; and (ii) Inhibitors of gap junction channels (GJCs) have confounding actions on EC electrical and Ca2+ signalling. EXPERIMENTAL APPROACH Intact EC tubes were isolated from abdominal muscle feed (superior epigastric) arteries of C57BL/6 mice. Hyperpolarization was initiated with indirect (ACh) and direct (NS309) stimulation of intermediate‐ and small‐conductance Ca2+‐activated K+ channels (IKCa/SKCa). Remote membrane potential (Vm) responses to intracellular current injection defined the length constant (λ) for electrical conduction. Dye coupling was evaluated following intracellular microinjection of propidium iodide. Intracellular Ca2+ dynamics were determined using Fura‐2 photometry. Carbenoxolone (CBX) or β‐glycyrrhetinic acid (βGA) was used to investigate the role of GJCs. KEY RESULTS Steady‐state Vm of ECs was −25 mV. ACh and NS309 hyperpolarized ECs by −40 and −60 mV respectively. Electrical conduction decayed monoexponentially with distance (λ∼1.4 mm). Propidium iodide injected into one EC spread into surrounding ECs. CBX or βGA inhibited dye transfer, electrical conduction and EC hyperpolarization reversibly. Both agents elevated resting Ca2+ while βGA inhibited responses to ACh. CONCLUSIONS AND IMPLICATIONS Individual cells were effectively coupled to each other within EC tubes. Inhibiting GJCs with glycyrrhetinic acid derivatives blocked hyperpolarization mediated by IKCa/SKCa channels, regardless of Ca2+ signalling, obviating use of these agents in distinguishing key determinants of electrical conduction along the endothelium.