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Analysis of type I and IV collagens by FT-IR spectroscopy and imaging for a molecular investigation of skeletal muscle connective tissue

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Abstract

Many muscular diseases result from abnormal organization of connective tissue and/or collagen network formation. Only a few molecular imaging techniques are able to analyze this collagen network by differentiating collagen types. In this study, FT-IR spectroscopy was used to analyze type I and IV collagens, the most important compounds of which are perimysium and endomysium, respectively. Secondary structure of collagen types was determined by curve-fitting the 1,700–1,480 cm−1 spectral interval. Type I collagen could be differentiated from type IV by its higher amounts of triple helix and α-helix, but lower amounts of β-sheets (P < 0.01). FT-IR imaging was then used to determine structural features of perimysium and endomysium collagen network in bovine Flexor carpi radialis muscle. Secondary structure of proteins contained in perimysium and endomysium was found to be very close to type I and IV collagens, respectively. FT-IR spectroscopy and imaging are thus analytical tools that might be used for investigating biodistribution and assembly of collagen types in connective tissues.

Visible (left) and full spectral FT-IR (right) images of skeletal muscle tissue section (16 μm) exhibiting a vertical arrangement of fibers. + and × in FT-IR image show selected positions to obtain FT-IR spectra of perimysium and endomysium, respectively

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Abbreviations

ECM:

extracellular matrix

FT-IR:

Fourier-transform infrared

IMCT:

intramuscular connective tissue

tI-C:

type I collagen

tIV-C:

type IV collagen

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Authors and Affiliations

  1. CNRS UMR 5084, CNAB, Bio-Organic Chemistry Group, Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076, Bordeaux Cedex, France

    Cyril Petibois, Gilles Gouspillou, Katia Wehbe & Gérard Déléris

  2. INSERM U688 Physiopathologie Mitochondriale, Université Victor Segalen Bordeaux 2, 146 rue Léo Saignat, 33076, Bordeaux Cedex, France

    Gilles Gouspillou & Jean-Paul Delage

Authors
  1. Cyril Petibois
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  2. Gilles Gouspillou
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  3. Katia Wehbe
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  4. Jean-Paul Delage
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  5. Gérard Déléris
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Correspondence to Cyril Petibois.

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Petibois, C., Gouspillou, G., Wehbe, K. et al. Analysis of type I and IV collagens by FT-IR spectroscopy and imaging for a molecular investigation of skeletal muscle connective tissue. Anal Bioanal Chem 386, 1961–1966 (2006). https://doi.org/10.1007/s00216-006-0828-0

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  • DOI: https://doi.org/10.1007/s00216-006-0828-0

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