Structural dynamics of the skeletal muscle fiber by second harmonic generation

V. Nucciotti; C. Stringari; L. Sacconi; F. Vanzi; M. Linari; G. Piazzesi; V. Lombardi; F. S. Pavone

doi:10.1117/12.762704

15 February 2008 Structural dynamics of the skeletal muscle fiber by second harmonic generation

V. Nucciotti, C. Stringari, L. Sacconi, F. Vanzi, M. Linari, G. Piazzesi, V. Lombardi, F. S. Pavone

Proceedings Volume 6860, Multiphoton Microscopy in the Biomedical Sciences VIII; 68600A (2008) https://doi.org/10.1117/12.762704
Event: SPIE BiOS, 2008, San Jose, California, United States

Abstract

The high degree of structural order in skeletal muscle allows imaging of this tissue by Second Harmonic Generation (SHG). As previously found (Vanzi et al., J. Muscle Cell Res. Motil. 2006) by fractional extraction of proteins, myosin is the source of SHG signal. A full characterization of the polarization-dependence of the SHG signal can provide very selective information on the orientation of the emitting proteins and their dynamics during contraction. We developed a line scan polarization method, allowing measurements of a full polarization curve in intact muscle fibers from skeletal muscle of the frog to characterize the SHG polarization dependence on different physiological states (resting, rigor and isometric tetanic contraction). The polarization data have been interpreted by means of a model in terms of the average orientation of SHG emitters.The different physiological states are characterized by distinct patterns of SHG polarization. The variation of the orientation of emitting molecules in relation to the physiological state of the muscle demonstrates that one part of SHG signal arises from the globular head of the myosin molecule that cross-links actin and myosin filaments. The dependence of the SHG modulation on the degree of overlap between actin and myosin filaments during an isometric contraction, provides the constraints to estimate the fraction of myosin heads generating the isometric force in the active muscle fiber.

Citation Download Citation

V. Nucciotti, C. Stringari, L. Sacconi, F. Vanzi, M. Linari, G. Piazzesi, V. Lombardi, and F. S. Pavone "Structural dynamics of the skeletal muscle fiber by second harmonic generation", Proc. SPIE 6860, Multiphoton Microscopy in the Biomedical Sciences VIII, 68600A (15 February 2008); https://doi.org/10.1117/12.762704

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KEYWORDS

Second-harmonic generation

Molecules

Polarization

Data modeling

Head

Harmonic generation

Tissues

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