title: Cellular uptake mechanisms of 99mTcN-NOET in cardiomyocytes from newborn rats: calcium channel interaction.
abstract: BACKGROUND
  Bis[N-ethoxy,N-ethyl(dithiocarbamato)]nitrido Tc (V) (TcN-NOET) is a new technetium complex proposed as a tracer of myocardial perfusion. However, its cellular uptake mechanisms are unknown, although membrane localization on rat heart preparations and preferential binding to polymorphonuclear neutrophils (PMNs) have been reported. Because of the central role of calcium in PMN actions, a relationship was hypothesized between this ion flux and TcN-NOET cellular uptake.
  
  
  RESULTS
  The mechanisms of cellular uptake of TcN-NOET were investigated in newborn rat cardiomyocytes by study of the effect of calcium channel modulators on tracer binding. Nifedipine had no effect on tracer uptake at 1 minute. However, verapamil 0.1 micromol/L and diltiazem 0.5 micromol/L induced a 40% decrease in uptake. Conversely, Bay K 8644 0.25 micromol/L increased TcN-NOET uptake by 73%. Alterations in other membrane ion transports failed to modify tracer uptake, indicating the specificity of the relationship between TcN-NOET uptake and calcium channels. Kinetic studies indicated that cellular net accumulation of the tracer was slow (t1/2=28.5 minutes) and retention was prolonged (84% of initial activity retained after 120 minutes of washout). The energy dependence of TcN-NOET uptake was investigated after 60 minutes of metabolic inhibition by iodoacetic acid plus rotenone. The ATP decrease was not associated with reduction in tracer uptake at 1 minute (114.9+/-21.9% of control, P=NS).
  
  
  CONCLUSIONS
  The decrease in uptake observed with verapamil and diltiazem, the increase with Bay K 8644, and the lack of effect with nifedipine suggest that TcN-NOET binds to L-type calcium channels in the open configuration, without entering cardiomyocytes. The kinetics of TcN-NOET accumulation and retention are slow, and the mechanism for cellular uptake is not energy-dependent. From a clinical point of view, the effect of concurrent treatment by calcium inhibitors on myocardial binding of TcN-NOET should be taken into account.
authors.affiliation: Laboratoire d'Etudes des Radiopharmaceutiques, ESA CNRS 5077, Faculté de médecine, Université Joseph Fourier, Grenoble, France.
authors.forename: L
authors.initials: L
authors.lastname: Riou
authors.forename: C
authors.initials: C
authors.lastname: Ghezzi
authors.forename: O
authors.initials: O
authors.lastname: Mouton
authors.forename: J P
authors.initials: JP
authors.lastname: Mathieu
authors.forename: R
authors.initials: R
authors.lastname: Pasqualini
authors.forename: M
authors.initials: M
authors.lastname: Comet
authors.forename: D
authors.initials: D
authors.lastname: Fagret
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D015220:Calcium Channels / Q000378:metabolism*; D009206:Myocardium / Q000378:metabolism*; D015609:Organotechnetium Compounds / Q000378:metabolism* / Q000493:pharmacokinetics; D051381:Rats; D013859:Thiocarbamates / Q000378:metabolism* / Q000493:pharmacokinetics
chemical_list: D015220:Calcium Channels; D015609:Organotechnetium Compounds; D013859:Thiocarbamates; C093922:technetium Tc 99m, bis(N-ethoxy-N-ethyldithiocarbamato)nitrido-