Mitochondrial monoamine oxidase-A-mediated hydrogen peroxide generation enhances 5-hydroxytryptamine-induced contraction of rat basilar artery

Br J Pharmacol. 2010 Nov;161(5):1086-98. doi: 10.1111/j.1476-5381.2010.00941.x.

Abstract

Background and purpose: We evaluated the role(s) of monoamine oxidase (MAO)-mediated H₂O₂ generation on 5-hydroxytryptamine (5-HT)-induced tension development of isolated basilar artery of spontaneously hypertensive rats (SHR) and normotensive Wistar-Kyoto (WKY) rats.

Experimental approach: Basilar artery (endothelium-denuded) was isolated for tension measurement and Western blots. Enzymically dissociated single myocytes from basilar arteries were used for patch-clamp electrophysiological and confocal microscopic studies.

Key results: Under resting tension, 5-HT elicited a concentration-dependent tension development with a greater sensitivity (with unchanged maximum tension development) in SHR compared with WKY (EC(50) : 28.4 ± 4.1 nM vs. 98.2 ± 9.4 nM). The exaggerated component of 5-HT-induced tension development in SHR was eradicated by polyethylene glycol-catalase, clorgyline and citalopram whereas exogenously applied H₂O₂ enhanced the 5-HT-elicited tension development in WKY. A greater protein expression of MAO-A was detected in basilar arteries from SHR than in those from WKY. In single myocytes and the entire basilar artery, 5-HT generated (clorgyline-sensitive) a greater amount of H₂O₂ in SHR compared with WKY. Whole-cell iberiotoxin-sensitive Ca(2+) -activated K(+) (BK(Ca) ) amplitude measured in myocytes of SHR was approximately threefold greater than that in WKY (at +60 mV: 7.61 ± 0.89 pA·pF(-1) vs. 2.61 ± 0.66 pA·pF(-1) ). In SHR myocytes, 5-HT caused a greater inhibition (clorgyline-, polyethylene glycol-catalase- and reduced glutathione-sensitive) of BK(Ca) amplitude than in those from WKY.

Conclusions and implications: 5-HT caused an increased generation of mitochondrial H₂O₂ via MAO-A-mediated 5-HT metabolism, which caused a greater inhibition of BK(Ca) gating in basilar artery myocytes, leading to exaggerated basilar artery tension development in SHR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Basilar Artery / drug effects
  • Basilar Artery / metabolism
  • Blotting, Western
  • Dose-Response Relationship, Drug
  • Hydrogen Peroxide / metabolism*
  • Hypertension / physiopathology
  • Male
  • Microscopy, Confocal
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Monoamine Oxidase / metabolism*
  • Muscle Cells / drug effects
  • Muscle Cells / metabolism
  • Patch-Clamp Techniques
  • Peptides / pharmacology
  • Potassium Channels, Calcium-Activated / antagonists & inhibitors
  • Potassium Channels, Calcium-Activated / metabolism
  • Rats
  • Rats, Inbred SHR
  • Rats, Inbred WKY
  • Serotonin / administration & dosage
  • Serotonin / pharmacology*
  • Vasoconstriction / drug effects

Substances

  • Peptides
  • Potassium Channels, Calcium-Activated
  • Serotonin
  • iberiotoxin
  • Hydrogen Peroxide
  • Monoamine Oxidase