ABSTRACT
Two-dimensional maps of birefringence in sarcomers of a single fiber of rabbit m.psoas were obtained by an automated interference microscope developed at our laboratory. The changes in birefringence of muscle fibers reflect the movement of myosin cross-bridges. The orientation of cross-bridges was modified by varying the pH (pH 7.0, 6.0, 8.0) and ionic strength (mu = 0.115, 0.085, 0.235) of the bathing rigor solution. The maximum value of total birefringence in the rigor state was observed at neutral pH. Total birefringence markedly decreased (by 40%) as pH was changed from 7.0 to both 8.0 and 6.0. No significant changes in light phase shifts were found at a 1.5 reduction of ionic strength in the rigor solution. The calculated birefringence values were 45% higher in rigor solutions of a high (mu = 0.235) ionic strength. The results observed are discussed in terms of changes in the orientation of cross-bridges due to the movement of the alpha-helical subfragment-2 away from the filament shaft (pH 8) or coming closer to it (mu = 0.235). The available data do not allow one to explain the results obtained at pH 6.0.
Subject(s)
Muscle Fibers, Skeletal/chemistry , Myosins/chemistry , Animals , Birefringence , In Vitro Techniques , Interferometry , Muscle Fibers, Skeletal/physiology , Muscle Relaxation , Muscle, Skeletal/chemistry , Muscle, Skeletal/physiology , Muscle, Skeletal/ultrastructure , Rabbits , Sarcomeres/chemistryABSTRACT
A new method of automatized quantitative interferometry of skeletal muscle fibers was developed for the investigation of birefringence. A device based on the Linnic microscope was constructed to obtain phase images, which are two-dimensional pictures of birefringence. For the first time, two-and-three-dimensional maps of both total birefringence and birefringence for individual sarcomeres in the central part of muscle fiber were visualized using large databases. It was shown that total birefringence of fibers at rest length in the rigor state was lower as compared with the relaxed. Birefringence values from individual sarcomere interferograms revealed also that normalized A-disk birefringence was lower in the rigor state. The results obtained could be explained by a decrease of thick filaments anisotropy, due to the moving away of myosin heads from the rod during transition into the rigor state.