Abstract
A highly organized cytoskeleton architecture is the basis for continuous and controlled contraction in cardiomyocytes (CMs). Abnormalities in cytoskeletal elements, like the Z-disc, are linked to several diseases. It is challenging to reveal the mechanisms of CM failure, endogenous repair, or mechanical homeostasis on the scale of single cytoskeletal elements. Here, we used a femtosecond (fs) laser to ablate single Z-discs in human pluripotent stem cells (hPSC) -derived CMs (hPSC-CM) and neonatal rat CMs. We show, that CM viability was unaffected by the loss of a single Z-disc. Furthermore, more than 40% of neonatal rat and 68% of hPSC-CMs recovered the Z-disc loss within 24 h. Significant differences to control cells, after the Z-disc loss, in terms of cell perimeter, x- and y-expansion and calcium homeostasis were not found. Only 14 days in vitro old hPSC-CMs reacted with a significant decrease in cell area, x- and y-expansion 24 h past nanosurgery. This demonstrates that CMs can compensate the loss of a single Z-disc and recover a regular sarcomeric pattern during spontaneous contraction. It also highlights the significant potential of fs laser-based nanosurgery to physically micro manipulate CMs to investigate cytoskeletal functions and organization of single elements.
Highlights
The cellular cytoskeleton represents a complex ubiquitous element across all cell types
Cardiomyocyte viability is unaffected by loss of a single Z-disc
In an initial series of experiments, we determined the viability of CMs after ablation of single Z-disc in the sarcomeric cytoskeleton
Summary
The cellular cytoskeleton represents a complex ubiquitous element across all cell types It defines the shape, determines mechanical properties, and affects signaling pathways[1,2]. In response to increased calcium levels, the well-controlled cross-bridge cycle between actin and myosin filaments leads to force generation and muscle shortening[3,4] Single sarcomeres in this sarcomeric cytoskeleton are connected at the lateral ends via Z-discs. A tool to better identify the fundamental function of single Z-discs in a disease context versus appropriate controls is attractive to investigate respective disease mechanisms in a dish This may support studies on pharmacological treatments and elucidate their potential effect(s) on cell regeneration versus disruption thereof.
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