TY - JOUR
T1 - Electrical pacing counteracts intrinsic shortening of action potential duration of neonatal rat ventricular cells in culture
AU - Sathaye, Alok
AU - Bursac, Nenad
AU - Sheehy, Sean
AU - Tung, Leslie
N1 - Funding Information:
We are grateful to Dr. Paul McDermott for his advice on the stimulation chamber design, and to Dr. Gordon Tomaselli for his assistance with the immunohistochemical portion of this study. This work was supported by NIH R01 HL66239.
PY - 2006/10
Y1 - 2006/10
N2 - Previous studies have demonstrated the relationship between the functional electrophysiological properties of cultured neonatal rat ventricular myocytes (NRVMs) and the ability of the substrate to induce and sustain arrhythmia. The goal of this study was to examine the effects of chronic pacing at a constant rate akin to that in vivo, on the functional electrophysiological properties of NRVM monolayers. Confluent NRVM monolayers grown on 20 mm diameter cover slips were left either unpaced or were stimulated at 3 Hz for the duration of the culture, and were optically mapped on days 4, 6, or 8. Action potential duration at 80% repolarization (APD80), conduction velocity (CV), and Kv4.3 (Ito) and NCX protein expression were measured. The effects of the excitation-contraction uncoupler 2,3-butadione monoxime (BDM) were also investigated. The 2 Hz APD80 of non-paced monolayers decreased significantly on days 6 (137.1 ± 13.9 ms) and 8 (109.8 ± 9.0 ms) compared with day 4 (197.0 ± 11.8 ms), while that of paced monolayers did not (206.8 ± 9.7, 209.1 ± 9.2, and 210.6 ± 9.9 ms, respectively). The 2 Hz CV of non-paced monolayers increased significantly on days 6 (26.0 ± 1.6 cm/s) and 8 (26.5 ± 1.0 cm/s) compared with day 4 (20.0 ± 1.0 cm/s), while that of paced monolayers did not change significantly (26.0 ± 2.0, 26.0 ± 1.0, and 23.8 ± 1.2 cm/s, respectively). The restitution curves of APD80 and CV of paced monolayers were also unchanging from days 4 through 8. Despite the unchanging APD80 and CV, a decrease in Kv4.3 expression and an increase in NCX expression were observed in paced compared with non-paced monolayers. Cessation of pacing or administration of BDM caused a reversal of phenotype back to that of non-paced monolayers. In summary, chronic electrical stimulation of confluent NRVM monolayers results in stabilization of APD80 and an advancement of the developmental rise of CV that is mediated by electromechanical coupling. These effects produce a steadier functional phenotype that may be beneficial for electrophysiological studies.
AB - Previous studies have demonstrated the relationship between the functional electrophysiological properties of cultured neonatal rat ventricular myocytes (NRVMs) and the ability of the substrate to induce and sustain arrhythmia. The goal of this study was to examine the effects of chronic pacing at a constant rate akin to that in vivo, on the functional electrophysiological properties of NRVM monolayers. Confluent NRVM monolayers grown on 20 mm diameter cover slips were left either unpaced or were stimulated at 3 Hz for the duration of the culture, and were optically mapped on days 4, 6, or 8. Action potential duration at 80% repolarization (APD80), conduction velocity (CV), and Kv4.3 (Ito) and NCX protein expression were measured. The effects of the excitation-contraction uncoupler 2,3-butadione monoxime (BDM) were also investigated. The 2 Hz APD80 of non-paced monolayers decreased significantly on days 6 (137.1 ± 13.9 ms) and 8 (109.8 ± 9.0 ms) compared with day 4 (197.0 ± 11.8 ms), while that of paced monolayers did not (206.8 ± 9.7, 209.1 ± 9.2, and 210.6 ± 9.9 ms, respectively). The 2 Hz CV of non-paced monolayers increased significantly on days 6 (26.0 ± 1.6 cm/s) and 8 (26.5 ± 1.0 cm/s) compared with day 4 (20.0 ± 1.0 cm/s), while that of paced monolayers did not change significantly (26.0 ± 2.0, 26.0 ± 1.0, and 23.8 ± 1.2 cm/s, respectively). The restitution curves of APD80 and CV of paced monolayers were also unchanging from days 4 through 8. Despite the unchanging APD80 and CV, a decrease in Kv4.3 expression and an increase in NCX expression were observed in paced compared with non-paced monolayers. Cessation of pacing or administration of BDM caused a reversal of phenotype back to that of non-paced monolayers. In summary, chronic electrical stimulation of confluent NRVM monolayers results in stabilization of APD80 and an advancement of the developmental rise of CV that is mediated by electromechanical coupling. These effects produce a steadier functional phenotype that may be beneficial for electrophysiological studies.
KW - Cell culture
KW - Chronic pacing
KW - Monolayers
KW - Neonatal rat ventricular myocyte
KW - Optical mapping
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U2 - 10.1016/j.yjmcc.2006.06.076
DO - 10.1016/j.yjmcc.2006.06.076
M3 - Article
C2 - 16950369
AN - SCOPUS:33748542565
SN - 0022-2828
VL - 41
SP - 633
EP - 641
JO - Journal of Molecular and Cellular Cardiology
JF - Journal of Molecular and Cellular Cardiology
IS - 4
ER -