High-resolution imaging of muscle attachment structures in Caenorhabditis elegans

Hiroshi Qadota, Yohei Matsunaga, Ken C.Q. Nguyen, Alexa Mattheyses, David H. Hall, Guy M. Benian

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We used structured illumination microscopy (SIM) to obtain super-resolution images of muscle attachment structures in Caenorhabditis elegans striated muscle. SIM imaging of M-line components revealed two patterns: PAT-3 (β-integrin) and proteins that interact in a complex with the cytoplasmic tail of β-integrin and localize to the basal muscle cell membrane [UNC-112 (kindlin), PAT-4 (ILK), UNC-97 (PINCH), PAT-6 (α-parvin), and UNC-95], are found in discrete, angled segments with gaps. In contrast, proteins localized throughout the depth of the M-line (UNC-89 (obscurin) and UNC-98) are imaged as continuous lines. Systematic immunostaining of muscle cell boundaries revealed that dense body components close to the basal muscle cell membrane also localize at cell boundaries. SIM imaging of muscle cell boundaries reveal "zipper-like" structures. Electron micrographs reveal electron dense material similar in appearance to dense bodies located adjacent to the basolateral cell membranes of adjacent muscle cells separated by ECM. Moreover, by EM, there are a variety of features of the muscle cell boundaries that help explain the zipper-like pattern of muscle protein localization observed by SIM. Short dense bodies in atn-1 mutants that are null for α-actinin and lack the deeper extensions of dense bodies, showed "zipper-like" structures by SIM similar to cell boundary structures, further indicating that the surface-proximal components of dense bodies form the "zipper-like" structures at cell boundaries. Moreover, mutants in thin and thick filament components do not have "dot-like" dense bodies, suggesting that myofilament tension is required for assembly or maintenance of proper dense body shape.

Original languageEnglish (US)
JournalCytoskeleton
DOIs
StateAccepted/In press - 2017

Fingerprint

Caenorhabditis elegans
Muscle Cells
Lighting
Microscopy
Muscles
Cell Membrane
Integrins
Electrons
Actinin
Myeloma Proteins
Striated Muscle
Muscle Proteins
Myofibrils
Tail
Proteins
Maintenance

Keywords

  • C. elegans
  • Electron microscopy
  • Integrin adhesion sites
  • Muscle
  • Super resolution microscopy

ASJC Scopus subject areas

  • Structural Biology
  • Cell Biology

Cite this

High-resolution imaging of muscle attachment structures in Caenorhabditis elegans. / Qadota, Hiroshi; Matsunaga, Yohei; Nguyen, Ken C.Q.; Mattheyses, Alexa; Hall, David H.; Benian, Guy M.

In: Cytoskeleton, 2017.

Research output: Contribution to journalArticle

Qadota, Hiroshi ; Matsunaga, Yohei ; Nguyen, Ken C.Q. ; Mattheyses, Alexa ; Hall, David H. ; Benian, Guy M. / High-resolution imaging of muscle attachment structures in Caenorhabditis elegans. In: Cytoskeleton. 2017.
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AB - We used structured illumination microscopy (SIM) to obtain super-resolution images of muscle attachment structures in Caenorhabditis elegans striated muscle. SIM imaging of M-line components revealed two patterns: PAT-3 (β-integrin) and proteins that interact in a complex with the cytoplasmic tail of β-integrin and localize to the basal muscle cell membrane [UNC-112 (kindlin), PAT-4 (ILK), UNC-97 (PINCH), PAT-6 (α-parvin), and UNC-95], are found in discrete, angled segments with gaps. In contrast, proteins localized throughout the depth of the M-line (UNC-89 (obscurin) and UNC-98) are imaged as continuous lines. Systematic immunostaining of muscle cell boundaries revealed that dense body components close to the basal muscle cell membrane also localize at cell boundaries. SIM imaging of muscle cell boundaries reveal "zipper-like" structures. Electron micrographs reveal electron dense material similar in appearance to dense bodies located adjacent to the basolateral cell membranes of adjacent muscle cells separated by ECM. Moreover, by EM, there are a variety of features of the muscle cell boundaries that help explain the zipper-like pattern of muscle protein localization observed by SIM. Short dense bodies in atn-1 mutants that are null for α-actinin and lack the deeper extensions of dense bodies, showed "zipper-like" structures by SIM similar to cell boundary structures, further indicating that the surface-proximal components of dense bodies form the "zipper-like" structures at cell boundaries. Moreover, mutants in thin and thick filament components do not have "dot-like" dense bodies, suggesting that myofilament tension is required for assembly or maintenance of proper dense body shape.

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