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Author Notes:

Hang Lu, School of Chemical and Biomolecular Engineering, Georgia Institute of Technology, Atlanta, GA 30332, United States, hang.lu@gatech.edu

Guy M. Benian Department of Pathology, Emory University, Atlanta, GA 30322, United States, pathgb@emory.edu

The authors are grateful to Dr. David Garton (Georgia Tech) for his expert advice on statistical analysis.

Subjects:

Research Funding:

Some strains used in this work were provided by the Caenorhabditis Genetics Center, which is supported by the National Center for Research Resources of the NIH.

Other strains were obtained from Dr. Shohei Mitani at Tokyo Women’s Medical University School of Medicine. J. F. N. was supported by an REU from the National Science Foundation.

G.M.B. thanks the Emory University Research Committee; the Emory University Department of Pathology; and the American Heart Association for financial support.

H.L. thanks NIH, NSF and the Sloan Foundation for financial support.

Keywords:

  • Science & Technology
  • Life Sciences & Biomedicine
  • Biochemical Research Methods
  • Biochemistry & Molecular Biology
  • Muscle
  • Sarcomere
  • Muscle focal adhesions
  • Caenorhabditis elegans
  • Muscle genes
  • Locomotion assay
  • NEMATODE CAENORHABDITIS-ELEGANS
  • PROTEIN-KINASE DOMAINS
  • ALPHA-ACTININ
  • FILAMENT ORGANIZATION
  • THICK FILAMENTS
  • FINGER PROTEIN
  • M-LINES
  • COMPLEXES
  • UNC-98
  • UNC-97/PINCH

Bending amplitude - A new quantitative assay of C. elegans locomotion: Identification of phenotypes for mutants in genes encoding muscle focal adhesion components

Tools:

Journal Title:

Methods

Volume:

Volume 56, Number 1

Publisher:

, Pages 95-102

Type of Work:

Article | Post-print: After Peer Review

Abstract:

The nematode Caenorhabditis elegans uses striated muscle in its body wall for locomotion. The myofilament lattice is organized such that all the thin filament attachment structures (dense bodies, analogous to Z-disks) and thick filament organizing centers (M-lines) are attached to the muscle cell membrane. Thus, the force of muscle contraction is transmitted through these structures and allows locomotion of the worm. Dense bodies and M-lines are compositionally similar to focal adhesions and costameres, and are based on integrin and associated proteins. Null mutants for many of the newly discovered dense body and M-line proteins do not have obvious locomotion defects when observed casually, or when assayed by counting the number of times a worm moves back and forth in liquid. We hypothesized that many of these proteins, located as they are in muscle focal adhesions, function in force transmission, but we had not used an appropriate or sufficiently sensitive assay to reveal this function. Recently, we have developed a new quantitative assay of C. elegans locomotion that measures the maximum bending amplitude of an adult worm as it moves backwards. The assay had been used to reveal locomotion defects for null mutants of genes encoding ATN-1 (α-actinin) and PKN-1 (protein kinase N). Here, we describe the details of this method, and apply it to 21 loss of function mutants in 17 additional genes, most of which encode components of muscle attachment structures. As compared to wild type, mutants in 11 genes were found to have less ability to bend, and mutants in one gene were found to have greater ability to bend. Loss of function mutants for eight proteins had been reported to have normal locomotion (ZYX-1 (zyxin), ALP-1 (Enigma), DIM-1, SCPL-1), or locomotion that was not previously investigated (FRG-1 (FRG1), KIN-32 (focal adhesion kinase), LIM-8), or had only slightly decreased locomotion (PFN-3 (profilin)).

Copyright information:

© 2011 Elsevier Inc..

This is an Open Access work distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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