High selectivity of calcium channels in single dialysed heart cells of the guinea‐pig.

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Abstract
Membrane currents and action potentials were recorded in single ventricular cells obtained from guinea‐pig hearts by enzymatic dissociation. Ca2+ channel currents carried by Ba2+ or Ca2+ were recorded with a suction pipette (5‐10 microns diameter) for voltage clamp and internal dialysis. Currents through Na+, K+ and non‐selective monovalent cation channels were suppressed by suitable holding potentials and external and internal solutions. The dialysis method allowed exchange within minutes of alkali metal cations (e.g. Cs+) and small molecules (e.g. quaternary derivatives of lidocaine and verapamil). Nevertheless, Ca2+ channels remained functional for considerable periods, typically 20 min and sometimes more than 1 h. With Ba2+ outside and Cs+ inside, current flow through Ca2+ channels changed from inward to outward at strongly positive levels beyond a clear‐cut reversal potential Erev. Several methods for defining Erev were in close agreement: (1) zero‐crossing of leak‐subtracted peak current, (2) inversion of time‐dependent current changes during channel activation or inactivation, (3) inversion of drug‐sensitive current as defined by channel blockers such as Cd2+ or D‐600. Erev varied with external Ba2+ or internal Cs+. Erev increased by 29 mV per 10‐fold increase in Ba2+. Interpreted with constant‐field theory, Erev values correspond to PBa/PCs of approximately 1360. With 5 mM‐Ca2+ outside and 151 mM‐Cs+ inside, Ca2+ channel current reversed near + 75 mV, corresponding to PCa/PCs approximately 6000. Earlier measurements of Erev (Lee & Tsien, 1982) suggest that PCa/PK greater than 1000. At strongly positive membrane potentials where channel activation is maximal, the Ca2+ channel current‐voltage relationship is strongly non‐linear, with conductance increasing on either side of an inflexion point near Erev. Activation of inward or outward currents through Ca2+ channels follows a sigmoid time course, as expected if activation were a multi‐step process.