Visualization of cardiac muscle thin filaments and measurement of their lengths by electron tomography

Aims An intriguing difference between vertebrate skeletal and cardiac muscles is that the lengths of the thin filaments are constant in the former but variable in the tatter. The thick filaments have constant lengths in both types of muscles. The contractile behaviour of a muscle is affected by the Lengths of both types of filaments as the tension generated during contraction depends on the amount of filament overlap. To understand the behaviour of cardiac muscle, it is important to know the distribution of the thin filament lengths. The previous detailed analysis by Robinson and Winegrad used serial transverse sections to determine the lengths of the thin filaments. However, the precision, set by the 100 nm section thickness, was low. Here, we have used electron tomography to produce 3D images of rat and mouse cardiac muscles in which we can actually see individual thin filaments up to the free ends and see that these free ends have variable locations. For comparison, we also measure the thin filament Lengths in skeletal muscle (frog sartorius). Methods and results Cardiac papillary muscles were obtained from a rat (Sprague-Dawley) and a mouse (C57/B6). Skeletal muscle (sartorius) was obtained from a frog (Rana pipiens). Longitudinal sections (100 nm thick) were used to produce tilt series and tomograms from which the thin filament paths were traced. Cardiac papillary muscle thin filaments in rat and mouse range from 0.94 to 1.10 mu m, with a mean length of 1.04 mu m and standard deviation of 0.03 mu m. For frog sartorius muscle, the thin filament length was 0.94 mu m with standard deviation of 0.01 mu m. Conclusion Electron tomography of cardiac and skeletal muscles allows direct visualization and high precision measurement of the lengths of thin filaments.