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

The Histochemical Journal Pub Date : 1995-09-01
J A Santána Pereira, A de Haan, A Wessels, A F Moorman, A J Sargeant
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Abstract

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. Immunohistochemical 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).

大鼠IIX型纤维的mATPase组织化学特征:与肌球蛋白重链免疫标记的相关性。
在本研究中,我们报告了一种新的组织化学方法,该方法通过在碱性和酸性培养基中连续预孵育,根据肌纤维肌动球蛋白atp酶(mATPase)活性的特定谱选择性地区分表达肌球蛋白重链IIX的肌纤维。用抗肌球蛋白重链单克隆抗体检测酶反应的特异性,该抗体在肌肉匀浆的Western blots上进行表征。将传统pH缓冲液的酶组织化学反应与新方法的酶组织化学反应进行比较,并结合免疫反应,用于确认MyHC表达与mATPase不同谱之间的关系。免疫组化反应表明,新方法仅能区分表达肌球蛋白重链IIX的纤维。该方法揭示了一个连续体,其中中间染色强度对应于表达肌球蛋白重链IIX与IIA或IIB形式相结合的杂交纤维。定量组织化学和免疫组织化学(通过图像分析)用于检查mATPase染色强度与肌球蛋白重链IIX表达量之间的关系,表明新方法可以很好地区分表达可变量IIX亚型的杂交纤维(r2 = 0.93)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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