We directly tested the effects of nitric oxide (NO) on Na+ channels in guin
ea pig and mouse ventricular myocytes using patch-clamp recordings. We have
previously shown that NO donors have no observed effects on expressed Nachannels. In contrast, NO (half-blocking concentration of 523 nmol/L) signi
ficantly reduces peak whole-cell Na+ current (I-Na) in isolated ventricular
myocytes. The inhibitory effect of NO on I-Na was not associated with chan
ges in activation, inactivation, or reactivation kinetics. At the single-ch
annel level, the reduction in macroscopic current was mediated by a decreas
e in open probability and/or a decrease in the number of functional channel
s with no change in single-channel conductance. Application of cell permeab
le analogs of cGMP or CAMP mimics the inhibitory effects of NO. Furthermore
, the effects of NO on I-Na can only be blocked by inhibition of both cGMP
and CAMP pathways. Sulfhydryl-reducing, agent does not reverse the effect o
f NO. In summary, although NO exerts its action via the known guanylyl cycl
ase (GC)/cGMP pathway, our findings provide evidence that NO can mediate it
s function via a GC/cGMP-independent mechanism involving the activation of
adynylyl cyclase (AC) and cAMP-dependent protein kinase.