Introduction
: An important role for the inwardly rectifying potassium current (I
K1
) has been postulated in controlling the stability and frequency of rotors responsible for ventricular tachycardia and fibrillation. We investigated the effects of Kir2.1 overexpression and Kir2.1AAA dominant-negative mutant suppression on the electrophysiology and inducibility, stability and frequency of spiral waves in an
in vitro
cardiac tissue model.
Methods/Results
: Neonatal rat ventricular myocytes (NRVMs) were transduced by lentiviral vectors encoding Kir2.1 or Kir2.1AAA. Immunostaining revealed Kir2.1 or mutant Kir2.1 protein overexpression and whole cell-clamp confirmed the predicted effects on I
K1
, resting potential, and action potential duration (APD
80
). Optical mapping was performed on confluent NRVM monolayers containing a 5 mm diameter central island of gene-modified NRVMs created by a stenciling technique. APs propagated with increased CV (25.1±2.7 cm/sec, n=7) and shortened APD
80
(73±11 msec, n=7) in islands of Kir2.1 overexpression, or decreased CV (13.1±1.1 cm/sec, n=7) and prolonged APD
80
(263±14 msec, n=7) in islands of Kir2.1AAA suppression, compared with normal CV and APD
80
of 19.2±0.4 cm/sec and 169±14 msec (n=7) in non-transduced islands. Reentry was initiated by rapid pacing. With Kir2.1 overexpression, reentrant waves anchored to the island and remained stable (89±15 minutes, n=3) with a frequency of 8±2 Hz. Superfusion with 0.5 mM BaCl
2
to block I
K1
slowed reentry to 1 Hz and terminated it shortly after initiation. NRVM monolayers with islands of Kir2.1AAA suppression (n=3) displayed rapid spontaneous activity. Rapid pacing of these monolayers initiated an unstable figure-of-eight reentry (n=3) that degraded into single and multi-armed spiral waves, anchored to varying parts of the island with a maximum frequency of 2±1 Hz. Importantly, no reentry could be initiated in monolayers with non-transduced islands (n=3).
Conclusion
: Functional reentrant waves induced by rapid pacing are anchored to islands of localized Kir2.1 overexpression whereas they drop in frequency and meander in islands of dominant-negative suppression of Kir2.1, confirming the importance of I
K1
for the stability of these waves in cardiac tissue.
A hybrid stimulation strategy for suppression of spiral waves in cardiac tissue