Contractile Properties of Cardiac Muscle

Permeabilized trabeculae (mouse) or thin strips (pig) were dissected and mounted between a force transducer (Aurora Scientific, model 400A) and a motor (Aurora Scientific, model 315C) using aluminum T-clips (Aurora Scientific).^{14 (link)} Sarcomere length was set to ≈2.3 µm for the experiments. Experiments were conducted in physiological solution (pH 7.0) at 15 °C (mouse) or 21 °C (pig) containing a range of negative log of calcium concentration (pCa; −log₁₀[Ca²⁺]) values from 9.0 to 4.0 with and without 1 μM Danicamtiv (MedChemExpress). Tissue was allowed to reach stead-state force (F) at each pCa. F-pCa curves were collected and analyzed with custom code using LabView software and fit to the Hill equation. The rate of tension redevelopment (k_tr) and high-frequency stiffness was measured at each pCa (details in the Supplemental Material).

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Kooiker K.B., Mohran S., Turner K.L., Ma W., Martinson A., Flint G., Qi L., Gao C., Zheng Y., McMillen T.S., Mandrycky C., Mahoney-Schaefer M., Freeman J.C., Costales Arenas E.G., Tu A.Y., Irving T.C., Geeves M.A., Tanner B.C., Regnier M., Davis J, & Moussavi-Harami F. (2023). Danicamtiv Increases Myosin Recruitment and Alters Cross-Bridge Cycling in Cardiac Muscle. Circulation Research, 133(5), 430-443.

Publication 2023

model 400A model 315C aluminum T-clips Danicamtiv

Aluminum Calcium Clips Mouse Physiological Sarcomere Tissue Trabeculae Transducer

Corresponding Organization :

Other organizations : University of Washington, Washington State University, Illinois Institute of Technology, Dalian Medical University, University of Kent

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Variable analysis

independent variables

Negative log of calcium concentration (pCa; −log10[Ca2+]) values from 9.0 to 4.0
Presence or absence of 1 μM Danicamtiv (MedChemExpress)

dependent variables

Steady-state force (F)
Rate of tension redevelopment (ktr)
High-frequency stiffness

control variables

Sarcomere length set to ≈2.3 μm
Physiological solution (pH 7.0) at 15 °C (mouse) or 21 °C (pig)

controls

Positive control: Experiments with varying pCa values
Negative control: Experiments with 1 μM Danicamtiv

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