Advertisement

Development of an Apparatus to Control Load by Electromagnet for a Motility System In Vitro

  • Takashi Watari
  • Akira Ono
  • Yoshiki Ishii
  • Huang Zhenli
  • Shinichi Miyake
  • Teizo Tsuchiya
Conference paper
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 538)

Abstract

We developed an electromagnet to perform quick changes in load in the motility system consisting of myosin molecules attached to a magnetizable bead and actin filaments. The electromagnet was combined with an inverted microscope and load could be quickly changed under optical observation. The magnetic field was generated by high electric current (6V, 0-125A) and the maximum field was 8,000 0e. The maximum force exerted on a bead was 80pN at 2.5mm distance from a magnet. The change in force was 0.48 % at the distance of 5.0 mm from the magnet when a bead moved longitudinally for 30 μ m. The time to change load was about 20 ms. The movements of a bead in water were recorded by video when step changes in magnetic field were applied and it was shown that a bead exactly followed the change in force. This apparatus is very much useful to analyze the transient changes in the movement of a bead, if the movement is relatively slow as in the interaction between actin and myosin from molluscan smooth muscle.

Keywords

Motility System Quick Change Single Muscle Fiber Myosin Molecule Actin Cable 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Civan, M. M. and Podolsky, R., 1966, Contraction kinetics of striated muscle fibres following quick changes in load. J.Physiol. 184: 511–5PubMedGoogle Scholar
  2. Edman, K. A. P.,1979, The velocity of unloaded shortening and its relation to sarcomere length and isometric force in vertebrate muscle fibres. J. Physiol. 291: 143–159.PubMedGoogle Scholar
  3. Edman, K. A. P. and Curtain, N. A.,2001, Synchronal oscillations of length and stiffness during loaded shortening of frog muscle fibres. J. Physiol. 534: 552–563.CrossRefGoogle Scholar
  4. Edman, K. A. P. and Tsuchiya T.,1996, Strain of passive elements during force enhancement by stretch in frog muscle fibres. J. Physiol. 490: 191–205.PubMedGoogle Scholar
  5. Hill, A. V.,1938, The heat of shortening and the dynamic constants of muscle. Proc. Roy. Soc. B126: 136–195.Google Scholar
  6. Huxley, A. F. and Simmons, R. M.,1971, Proposed mechanism of force generation in striated muscle. Nature 233: 533–538.PubMedCrossRefGoogle Scholar
  7. Ishii, N., Tsuchiya, T. and Sugi, H.,1997, An in vitro motility assay system retaining the steady-state characteristics of muscle fibres under positive and negative load. Biochim. Biophy Acta 1319: 155–162.CrossRefGoogle Scholar
  8. Nishizaka, T., Seo, R., Tadakura, H., Kinosita, K. Jr and Ishiwata, S.,2000, characterization of single rigor bonds: load dependence of lifetime and mechanical properties. Biophy. J. 79: 962–9CrossRefGoogle Scholar
  9. Shin, W. M, Gryczynski, Z., Kakowicz, J. R. and Spudich, J. A.,2000, A fret-based sensor reveals large ATP hydrolysis-induced conformational changes and three distinct states of the molecular motor myosin. Cell 102: 683–694.CrossRefGoogle Scholar
  10. Sugi, H. and Tsuchiya, T., 1981a,Isotonic velocity transients in frog muscle fibres following quick changes in load. J. Physiol. 319: 219–238.PubMedGoogle Scholar
  11. Sugi, H. and Tsuchiya, T., 1981b,Enhancement of mechanical performance in frog muscle fibres after quick increases in load. J. Physiol. 319: 239–252.PubMedGoogle Scholar
  12. Sugiura, S., Kobayakawa, N., Fujita, H., Yamashita, H., Momomura, S., Chaen, S., Omata, M. and Sugi, H.,1998, Comparison of unitary displacements and forces between 2 cardiac myosin isoforms by the optical trap technique; molecular basis for cardiac adaptation. Circ. Res 82: 1029–1034.PubMedCrossRefGoogle Scholar
  13. Tanaka, H., Homma, K., Iwane-Hikikoshi, A., Katayama, E., Ikebe, R., Saito, J., Yanagida, T. and Ikebe, M.,2002, The motor domain determines the large step of myosin V. Nature 415: 192–195.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2003

Authors and Affiliations

  • Takashi Watari
  • Akira Ono
  • Yoshiki Ishii
  • Huang Zhenli
  • Shinichi Miyake
  • Teizo Tsuchiya
    • 1
  1. 1.Department of Biology, Faculty of ScienceKobe UniversityNada-ku, KobeJapan

Personalised recommendations