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Biophys. J. BioFAST: First Published October 28, 2005. doi:10.1529/biophysj.105.071555
© 2005 by the Biophysical Society.

A more recent version of this article appeared on January 15, 2006.
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Characterization of the myosin-based source for second-harmonic generation from muscle sarcomeres

Sergey V. Plotnikov 1, Andrew C. Millard 1, Paul J. Campagnola 1 and William Mohler 2*

1 UConn Health Center
2 University of Connecticut Health Center

* To whom correspondence should be addressed. E-mail: wmohler{at}neuron.uchc.edu.

Submitted on July 27, 2005
Revised on August 19, 2005
Accepted on 5 October 2005


  Abstract
Several biologically important protein structures give rise to strong second-harmonic generation (SHG) in their native context. In addition to high-contrast optical sections of cells and tissues, SHG imaging can provide detailed structural information based on the physical constraints of the optical effect. In this study we characterize, by biochemical and optical analysis, the critical structures underlying SHG from the complex muscle sarcomere. SHG emission arises from domains of the sarcomere containing thick filaments, even within nascent sarcomeres of differentiating myocytes. SHG from isolated myofibrils is abolished by extraction of myosin, but is unaffected by removal or addition of actin filaments. Furthermore, the polarization-dependence of sarcomeric SHG is not affected by either the proportion of myosin head domains or the orientation of myosin heads. By fitting SHG polarization anisotropy readings to theoretical response curves, we find an orientation for the elemental harmonophore that corresponds well to the pitch of the myosin rod ?-helix along the thick filament axis. Taken together, these data indicate that myosin rod domains are the key structures giving SHG from striated muscle. This study should guide the interpretation of SHG contrast in images of cardiac and skeletal muscle tissue for a variety of biomedical applications.

Key Words: muscle, myofibril, myosin, nonlinear optics, sarcomere, second-harmonic generation




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Copyright © 2005 by the Biophysical Society.