title: Changes in extracellular pH mediate the chronotropic responses to L-arginine.
abstract: We have recently shown that exogenous nitric oxide (NO) elicits a positive chronotropic response by stimulating the hyperpolarization activated current, I(f).
  
  
  OBJECTIVE
  To examine whether L-arginine (L-Arg) can mimic the chronotropic effect of NO by enhancing its endogenous production.
  
  
  METHODS
  In spontaneously beating guinea pig atria we evaluated the heart rate (HR) response to increasing concentrations of L-Arg (1 mumol/l to 10 mmol/l), and compared it with that for D-Arg or L-lysine (L-Lys) (all in free base (FB) or hydrochloride (HCl) formulation).
  
  
  RESULTS
  L-ArgFB > 100 mumol/l caused a reversible dose-dependent increase in HR (peak effect +64 +/- 7 bpm at 10 mmol/l, P < 0.05, n = 8). However, a similar HR response occurred with D-ArgFB (n = 7) or L-LysFB (n = 6). All FB formulations increased the perfusate pH (peak [pH]o = 8.61 +/- 0.03). Although alkalinization can stimulate NO release from the endothelium, this is unlikely to have contributed to HR changes in our preparation, since neither NG-methyl-L-arginine, (100-500 mumol/l, which per se reduced HR by 8 +/- 1%, P < 0.05, n = 9) nor NO scavenging (fresh 5% red blood cells, n = 9) caused a rightward shift of the concentration-response curve to L-ArgFB. Furthermore, as opposed to FB formulations, L-ArgHCl, D-ArgHCl or L-LysHCl > 1 mmol/l significantly decreased HR and [pH]o (n = 17). The chronotropic effects of L-ArFB or L-ArgHCl were reproduced by changing [pH]o with NaOH (n = 8) or HCl (n = 7), whereas the HR increase with L-ArgFB was prevented by clamping [pH]o at 7.42 +/- 0.07 (n = 10).
  
  
  CONCLUSIONS
  In vitro, L-Arg can markedly affect HR through a pH-mediated, NO-independent mechanism. Our data show that the opposing changes in [pH]o induced by different formulations of L-Arg can importantly confound the assessment of the biological effects of this amino acid.
authors.affiliation: Department of Cardiovascular Medicine, John Radcliffe Hospital, Oxford, UK. piotr.musialek@clinical-medicine.ox.ac.uk
authors.forename: P
authors.initials: P
authors.lastname: Musialek
authors.forename: D J
authors.initials: DJ
authors.lastname: Paterson
authors.forename: B
authors.initials: B
authors.lastname: Casadei
mesh_terms: D000704:Analysis of Variance; D000818:Animals; D001120:Arginine / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004791:Enzyme Inhibitors; D004912:Erythrocytes / Q000378:metabolism; D005110:Extracellular Space / Q000378:metabolism*; D006168:Guinea Pigs; D006339:Heart Rate / Q000187:drug effects*; D006863:Hydrogen-Ion Concentration; D007536:Isomerism; D008239:Lysine / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors; D010477:Perfusion; D013268:Stimulation, Chemical; D019323:omega-N-Methylarginine / Q000494:pharmacology
chemical_list: D004791:Enzyme Inhibitors; D019323:omega-N-Methylarginine; D001120:Arginine; D019001:Nitric Oxide Synthase; D008239:Lysine