4-acetamido-4--isothiocyanatostilbene-2-2--disulfonic-acid and methanesulfonic-acid

The Cai(2+)-insensitive transient outward current, ilo was studied at 20-24 degrees C in rat ventricular myocytes with the whole cell recording patch-clamp technique. The current was recorded before and after replacement of chloride by methanesulfonate or aspartate or in the absence and the presence of chloride channel blockers, SITS or 9-anthracene carboxylic acid. In control conditions (in the presence of external divalent cations, Ca2+ and Cd2+, Cd2+ being used to suppress Ca2+ current), ilo inactivation was composed of a fast and a slow component. When methanesulfonate was substituted for external Cl-, the peak current decreased to a variable extent, but the inactivation of the remaining current was still composed of a fast and a slow component. In contrast, the inactivation of the difference current was well fitted by a single exponential. The time to peak of the difference current was shorter than that of the current recorded either in the absence or the presence of methanesulfonate. Both activation- and steady-state inactivation-voltage curves were either unchanged (n = 4) or shifted by a few mV (5.5 mV, n = 14) towards positive potentials when methanesulfonate was substituted for Cl-. The current remaining in methanesulfonate reversed at potentials closed to EK. The difference current was composed of a peak and a steady-state component. The peak was suppressed by 4-aminopyridine whereas the steady-state component was not. The peak was also suppressed when pipette solution contained Cs+ instead of K+ but was still present when the Hepes concentration in both external and pipette media was increased 5-fold (50 mM vs. 10 mM). When aspartate was substituted for Cl- or when 2 mM SITS was added to the external solution (in the absence of Ca2+ and Cd2+ because aspartate is known to chelate Ca2+ ions and possibly other divalent cations), ilo was reduced to a similar extent in the two cases and the difference current was composed of a peak (inactivation fitted by a single exponential) and a steady-state component. The SITS-sensitive transient current reversed at a potential close to ECl. When 5 mM 9-anthracene carboxylic acid was added to external solution (in the presence of Ca2+ and Cd2+), the peak of the difference current was similar to that observed when Cl- was substituted by methanesulfonate. The difference current resulting from the substitution of methanesulfonate for chloride was not changed when the pipette solution contained either 50 mM EGTA (

Topics: 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; Animals; Anthracenes; Aspartic Acid; Cells, Cultured; Chloride Channels; Chlorides; Heart Ventricles; Membrane Potentials; Mesylates; Myocardium; Patch-Clamp Techniques; Rats

The effects of a chloride-poor medium (methanesulfonate substituted) and a chloride transport inhibitor (SITS) on the outward delayed current and the tonic tension were studied on frog atrial trabeculae under voltage-clamp conditions. The outward delayed current decreased in low-chloride medium (10.5 mmol/l) or in the presence of SITS (2 mmol/l). The tonic tension increased in chloride-poor solution and decreased following SITS. The replacement of chloride by methanesulfonate enhanced the transient increase of tonic tension induced by low external sodium concentration while SITS reduced it. In the same conditions, the effect of the chloride-poor medium was abolished in the presence of SITS. The results showing an increase in Na-Ca exchange in low-chloride medium and a decrease by SITS are discussed assuming that changes in the inner negative charge density influenced the Na-Ca exchange mechanism; the influence of pHi variation are also considered.

Topics: 4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; Animals; Atrial Function; Biological Transport; Calcium; Chlorides; Heart Atria; Ion Channels; Mesylates; Myocardial Contraction; Rana esculenta; Sodium