ABSTRACT
The mechanical strength of sarcomere structures of skeletal muscle was studied by rupturing single myofibrils of rabbit psoas muscle by submicromanipulation techniques. Microbeads coated with alpha-actinin were attached to the surface of myofibrils immobilized to coverslip. By use of either optical tweezers or atomic force microscope, the attached beads were captured and detached from the myofibrils. During the detachment of the beads, the actin filaments bound specifically to the beads were peeled off from the bulk structures of myofibrils, thus rupturing the peripheral components of the myofibrils bound to the actin filaments. By analyzing the ruptures thus produced in various myofibril preparations, it was found that the sarcomere structure of myofibrils is maintained by numerous molecular components having the mechanical strength sufficient to sustain the contractile force produced by the actomyosin system. The present techniques could be applied to study the mechanical strength of cellular organelles containing actin filaments as their component.
Subject(s)
Micromanipulation/methods , Myofibrils/physiology , Sarcomeres/physiology , Actin Cytoskeleton/physiology , Actin Cytoskeleton/ultrastructure , Actinin , Animals , Biomechanical Phenomena , Electrophoresis, Polyacrylamide Gel , Microscopy, Atomic Force , Microspheres , Muscle Contraction/physiology , Myofibrils/ultrastructure , Psoas Muscles/physiology , Psoas Muscles/ultrastructure , Rabbits , Sarcomeres/ultrastructureABSTRACT
The motions of myosin filaments actively sliding along suspended actin filaments were studied. By manipulating a double-beam laser tweezers, single actin filaments were suspended between immobilized microbeads. When another beads coated with myosin filaments were dragged to suspended actin filaments, the beads instantly and unidirectionally slid along the actin filaments. The video image analysis showed that the beads slid at a velocity of ca. 3-5 microm/s accompanied with zigzag motions. When beads were densely coated with myosin filaments, the sliding motions became straight and smooth. The obtained results indicate that (1) during the sliding motions, the interaction between myosin heads and actin filaments is weak and susceptible to random thermal agitations, (2) the effects of thermal agitations to the sliding motions of myofilaments are readily suppressed by mechanical constraints imposed to the filaments, and (3) the active sliding force is produced almost in parallel to the filaments axis.
