The mechanoelectric feedback: a novel "calcium clamp" method, using tetanic contraction, for testing the role of the intracellular free calcium.
Mechanical perturbations affect the membrane action potential, a phenomenon denoted as the mechanoelectric feedback (MEF), and may elicit cardiac arrhythmias. Two plausible mechanisms were suggested to explain this phenomenon: (i) stretch-activated channels (SACs) within the cell membrane and (ii) modulation of the action potential by the intracellular Ca(2+) (the Calcium hypothesis). The intracellular Ca(2+) varies mainly due to the effects of the mechanical perturbations on the affinity of troponin for calcium. The present study concentrates on the unique experimental methods that allow differentiating between the effects of SAC and Ca(2+) on the action potential. This is achieved by controlling the sarcomere lengths (SLs) independently of the intracellular Ca(2+) concentration, in the intact fiber. A dedicated experimental setup allowed simultaneous measurements of the membrane potential and the mechanical performance (Force and SL). The action potential was measured by voltage-sensitive dye (Di-4-ANEPPS). The SL was measured by laser diffraction technique and was controlled by a fast servomotor. The intracellular Ca(2+) was controlled (calcium clamp) by imposing stable tetanic contractions at various extracellular calcium concentrations ([Ca(2+)](0)s). Tetanus was obtained by 8 Hz stimulation in the presence of cyclopiazonic acid (CPA) (30 muM). Isolated trabeculae from a rat's right ventricle were studied at different SLs and [Ca(2+)](0)s. The experimental data strongly support the calcium hypothesis. Although the action potential duration (APD) decreases at longer SL, the [Ca(2+)](0) has a significantly larger effect on the APD. The APD decreases as the [Ca(2+)](0) increases. Understanding the underlying mechanism opens new research avenues for the development of therapeutic modalities for cardiac arrhythmias.
Yaniv
Yael
Y
Department of Biomedical Engineering, Technion, IIT, Haifa 32000, Israel.
Levy
Carmit
C
Landesberg
Amir
A
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D040542:Mechanotransduction, Cellular; D051381:Rats; D017207:Rats, Sprague-Dawley; D013746:Tetany
D002118:Calcium