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The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling

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Summary

In the present study we report a novel histochemical method which, by sequential pre-incubations in alkaline and acidic media, selectively differentiates muscle fibres expressing myosin heavy chain IIX, on the basis of a specific profile for myofibrillar actomyosin ATPase (mATPase) activity. The enzyme reactions were tested for specificity by means of anti-myosin heavy chain monoclonal antibodies, which were characterized on Western blots of muscle homogenates. Enzyme histochemical reactions with the traditional pH buffers were compared to those of the new method and, in conjunction with the immunoreactions, used to confirm the relationship between MyHC expression and the distinct profiles for mATPase. Imrnunohistochemical reactions demonstrated that the new method only differentiates those fibres expressing myosin heavy chain IIX. The method revealed a continuum in which the intermediate staining intensities corresponded to hybrid fibres expressing myosin heavy chain IIX in combination with either the IIA or IIB forms. Quantitative histochemistry and immunohistochemistry (by image analysis), used to examine the relationship between staining intensities for mATPase and amounts of myosin heavy chain IIX expression, revealed that the new method discriminates well between hybrid fibres expressing variable amounts of the IIX isoform (r2 = 0.93).

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Authors and Affiliations

  1. Department of Muscle & Exercise Physiology, Vrije University, Amsterdam, The Netherlands

    José A. A. Sant'Ana Pereira, Arnold De Haan & Anthony J. Sargeant

  2. Department of Anatomy & Embryology, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands

    José A. A. Sant'Ana Pereira, Andy Wessels & Antoon F. M. Moorman

Authors
  1. José A. A. Sant'Ana Pereira
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  2. Arnold De Haan
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  3. Andy Wessels
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  4. Antoon F. M. Moorman
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  5. Anthony J. Sargeant
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Pereira, J.A.A.S., De Haan, A., Wessels, A. et al. The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling. Histochem J 27, 715–722 (1995). https://doi.org/10.1007/BF02388544

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  • DOI: https://doi.org/10.1007/BF02388544

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