Cardiomyocyte subdomain contractility arising from microenvironmental stiffness and topography


Cellular structure and function are interdependent. To understand this relationship in beating heart cells, individual neonatal rat ventricular myocytes (NRVMs) were analyzed one and 3 days after plating when cultured on different stiffness (100, 400 kPa) and surface structures (flat or \(15\,\upmu \hbox {m}\) high, \(15\,\upmu \hbox {m}\) diameter, microposts spaced \(75 \,\upmu \hbox {m}\) apart) manufactured from polydimethylsiloxane. Myofibril structure seen by immunohistochemistry was organized in three dimensions when NRVMs were attached to microposts. On day three, paxillin distribution near the post serving as cellular anchorage was quantified on both soft posts (12.04 % of total voxel count) and stiff posts (8.16 %). Living NRVMs were analyzed using line scans for sarcomeric shortening and shortening velocity, and traction force microscopy for surface stress and surface tension. One day after plating, NRVMs shortened more on soft posts (\(1.08\,\upmu \hbox {m}\) at \(4.75 \,\upmu \hbox {m}/\hbox {s}\)) compared to either soft flat (\(0.84 \,\upmu \hbox {m}\) at \(3.41 \,\upmu \hbox {m}/\hbox {s}\)), stiff posts (\(0.66 \,\upmu \hbox {m}\) at \(2.88 \,\upmu \hbox {m}/\hbox {s}\)) or stiff flat (\(0.48 \,\upmu \hbox {m}\) at \(1.44 \,\upmu \hbox {m}/\hbox {s}\)). NRVMs have decreased shortening and shortening velocity on soft posts (\(1.04 \,\upmu \hbox {m}\) at \(3.85 \,\upmu \hbox {m}/\hbox {s}\)) compared to soft flat (\(0.72 \,\upmu \hbox {m}\) at \(2.36 \,\upmu \hbox {m}/\hbox {s}\)) substrates. The surface stress and surface tension increased over time for both soft post (\(29.12\,\hbox {kN}/\hbox {m}^{2}\) and \(30.10\,\upmu \hbox {N}/\hbox {mm}\) to \(42.87\,\hbox {kN}/\hbox {m}^{2}\) and \(51.68 \,\upmu \hbox {N}/\hbox {mm}\)) and flat (\(15.36\,\hbox {kN}/\hbox {m}^{2}\) and \(19.00\,\upmu \hbox {N}/\hbox {mm}\) to \(32.87\,\hbox {kN}/\hbox {m}^{2}\) and \(37.38\,\upmu \hbox {N}/\hbox {mm}\)) substrates. Paxillin displacement during contraction on day three was significantly greater in NRVMs attached to soft posts \((1.39\,\upmu \hbox {m})\) compared to flat \((1.16\,\upmu \hbox {m})\) substrates. The volume and time creating four-dimensional data, interpreted by structural engineering theory, demonstrate subdomain structure is maintained by the counterbalance between the external load acting upon and the internal forces generated by the cardiomyocyte. These findings provide further insight into localized regulation of cellular mechanical function.

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We gratefully acknowledge Drs Shen Sun and Michael Cho for providing assistance with AFM experiments. Gratitude is also extended to Dr. Mark Sussman for providing paxillin-GFP used in these experiments. We also thank Dr. Matthew W. Curtis for his help to customize the method of adhering beads to the substratum surface for the traction force microscopy experiments. Support to conduct this research was provided by NIH NHLBI T32/HL07692, PO/HL62426.

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Correspondence to Brenda Russell.

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Broughton, K.M., Russell, B. Cardiomyocyte subdomain contractility arising from microenvironmental stiffness and topography. Biomech Model Mechanobiol 14, 589–602 (2015) doi:10.1007/s10237-014-0624-2

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  • 4D Imaging
  • Anisotropic elastic deformation
  • Kymograph
  • Shortening
  • Shortening velocity
  • Traction force
  • Surface stress
  • Surface tension