Altered pharmacologic responsiveness of reduced L-type calcium currents in myocytes surviving in the infarcted heart.

The pharmacologic responses of macroscopic L-type calcium channel currents to the dihydropyridine agonist, Bay K 8644, and beta-adrenergic receptor stimulation by isoproterenol were studied in myocytes enzymatically dissociated from the epicardial border zone of the arrhythmic 5-day infarcted canine heart (IZs). Calcium currents were recorded at 36 degrees to 37 degrees C using the whole cell, patch clamp method and elicited by applying step depolarizations from a holding potential of -40 mV to various test potentials for 250-msec duration at 8-second intervals. A Cs+ -rich and 10 mM EGTA-containing pipette solution and a Na+ -and K+ -free external solutions were used to isolate calcium currents from other contaminating currents. During control, peak ICa,L density was found to be significantly less in IZs (4.0 +/- 1.1 pA/pF) than in myocytes dispersed from the epicardium of the normal noninfarcted heart (NZs; 6.5 +/- 1.8 pA/pF). Bay K 8644 (1 micro M) significantly increased peak ICa,L density 3.5-fold above control levels in both NZs (to 22.5 +/- 6.2 pA/pF; n = 7) and IZs (to 12.8 +/- 3.0 pA/pF; n = 5), yet peak ICa,L density in the presence of drug was significantly less in IZs than NZs. The effects of Bay K 8644 on kinetics of current decay and steady-state inactivation relations of peak ICa,L were similar in the two cell types. In contrast, the response of peak L-type current density to isoproterenol (1 micro M) was significantly diminished in IZs compared to NZs regardless of whether Ba2+ or Ca2+ ions carried the current. Thus, these results indicate an altered responsiveness to beta-adrenergic stimulation in cells that survive in the infarcted heart. Furthermore, application of forskolin (1 micro M and 10 micro M) or intracellular cAMP (200 micro M), agents known to act downstream of the beta-receptor, also produced a smaller increase in peak IBa density in IZs versus NZs, suggesting that multiple defects exist in the beta-adrenergic signaling pathway of IZs. In conclusion, these studies illustrate that reduced macroscopic calcium currents of cells in the infarcted heart exhibit an altered pharmacologic profile that has important implications in the development of drugs for the diseased heart.