Visualization of cardiac muscle thin filaments and measurement of their lengths by electron tomography.
OBJECTIVE
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 latter. 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).


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 microm, with a mean length of 1.04 microm and standard deviation of 0.03 microm. For frog sartorius muscle, the thin filament length was 0.94 microm with standard deviation of 0.01 microm.


CONCLUSIONS
Electron tomography of cardiac and skeletal muscles allows direct visualization and high precision measurement of the lengths of thin filaments.
Burgoyne
Thomas
T
Molecular Medicine Section, National Heart and Lung Institute, Faculty of Medicine, Imperial College London, London, UK.
Muhamad
Farina
F
Luther
Pradeep K
PK
D000818:Animals; D002352:Carrier Proteins / Q000235:genetics; D003599:Cytoskeleton / Q000648:ultrastructure*; D007091:Image Processing, Computer-Assisted; D021621:Imaging, Three-Dimensional; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008854:Microscopy, Electron; D018482:Muscle, Skeletal / Q000648:ultrastructure; D010210:Papillary Muscles / Q000648:ultrastructure*; D011898:Ranidae; D051381:Rats; D017207:Rats, Sprague-Dawley; D012518:Sarcomeres / Q000648:ultrastructure*; D014054:Tomography / Q000379:methods*; D000099163:Myosin Binding Protein C
D002352:Carrier Proteins; D000099163:Myosin Binding Protein C