Structural and functional plasticity in long-term cultures of adult ventricular myocytes.
Cultured heart cells have long been valuable for characterizing biological mechanism and disease pathogenesis. However, these preparations have limitations, relating to immaturity in key properties like excitation-contraction coupling and β-adrenergic stimulation. Progressive attenuation of the latter is intimately related to pathogenesis and therapy in heart failure. Highly valuable would be a long-term culture system that emulates the structural and functional changes that accompany disease and development, while concurrently permitting ready access to underlying molecular events. Accordingly, we here produce functional monolayers of adult guinea-pig ventricular myocytes (aGPVMs) that can be maintained in long-term culture for several weeks. At baseline, these monolayers exhibit considerable myofibrillar organization and a significant contribution of sarcoplasmic reticular (SR) Ca(2+) release to global Ca(2+) transients. In terms of electrical signaling, these monolayers support propagated electrical activity and manifest monophasic restitution of action-potential duration and conduction velocity. Intriguingly, β-adrenergic stimulation increases chronotropy but not inotropy, indicating selective maintenance of β-adrenergic signaling. It is interesting that this overall phenotypic profile is not fixed, but can be readily enhanced by chronic electrical stimulation of cultures. This simple environmental cue significantly enhances myofibrillar organization as well as β-adrenergic sensitivity. In particular, the chronotropic response increases, and an inotropic effect now emerges, mimicking a reversal of the progression seen in heart failure. Thus, these aGPVM monolayer cultures offer a valuable platform for clarifying long elusive features of β-adrenergic signaling and its plasticity.
Department of Biomedical Engineering, The Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
Rosy
R
Joshi-Mukherjee
Ivy E
IE
Dick
Ting
T
Liu
Brian
B
O'Rourke
David T
DT
Yue
Leslie
L
Tung
D000200:Action Potentials; D000375:Aging; D000818:Animals; D002118:Calcium; D015220:Calcium Channels / Q000378:metabolism; D020013:Calcium Signaling; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D003600:Cytosol / Q000378:metabolism; D004558:Electric Stimulation; D056966:Excitation Contraction Coupling; D015726:Giant Cells / Q000378:metabolism; D006168:Guinea Pigs; D006329:Heart Conduction System / Q000502:physiology; D006352:Heart Ventricles / Q000166:cytology*; D008297:Male; D008954:Models, Biological; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology*; D009210:Myofibrils / Q000378:metabolism; D021122:Protein Subunits / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D013997:Time Factors
D015220:Calcium Channels; D021122:Protein Subunits; D011943:Receptors, Adrenergic, beta; D002118:Calcium
Adult cardiomyocyte; Cardiac electrophysiology; Cell culture; Electrical stimulation; Optical mapping; β-Adrenergic response