Medical Genetics Branch, National Human Genome Research Institute, National Institutes of Health, 10 Center Drive, Bld. 10, Room 10C103 Bethesda, Maryland, 20892, USA.
Therapeutics for Rare and Neglected Diseases, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, Maryland, USA.
Muscle Nerve. 2018 Aug;58(2):286-292. doi: 10.1002/mus.26135. Epub 2018 Apr 23.
GNE myopathy is an adult-onset muscle disorder characterized by impaired sialylation of (muscle) glycans, detectable by lectin histochemistry. We describe a standardized method to quantify (lectin-) fluorescence in muscle sections, applicable for diagnosis and response to therapy for GNE myopathy.
Muscle sections were fluorescently labeled with the sialic acid-binding Sambucus nigra agglutinin (SNA) lectin and antibodies to sarcolemma residence protein caveolin-3 (CAV-3). Entire tissue sections were imaged in tiles and fluorescence was quantified.
SNA fluorescence co-localizing with CAV-3 was ∼50% decreased in GNE myopathy biopsies compared with muscle-matched controls, confirming previous qualitative results.
This quantitative fluorescence method can accurately determine sialylation status of GNE myopathy muscle biopsies. This method is adaptable for expression of other membrane-associated muscle proteins, and may be of benefit for disorders in which therapeutic changes in expression are subtle and difficult to assess by other methods. Muscle Nerve 58: 286-292, 2018.
GNE 肌病是一种成人发病的肌肉疾病，其特征是（肌肉）糖链唾液酸化受损，可通过凝集素组织化学检测到。我们描述了一种标准化的方法来量化肌肉切片中的（凝集素）荧光，适用于 GNE 肌病的诊断和治疗反应。
肌肉切片用唾液酸结合的黑接骨木凝集素（SNA）凝集素和肌膜驻留蛋白窖蛋白-3（CAV-3）抗体进行荧光标记。整个组织切片以瓷砖形式成像，并对荧光进行定量。
与肌肉匹配的对照组相比，GNE 肌病活检中的 SNA 荧光与 CAV-3 共定位减少了约 50%，证实了之前的定性结果。
这种定量荧光方法可以准确确定 GNE 肌病肌肉活检的唾液酸化状态。该方法适用于其他膜相关肌肉蛋白的表达，对于其他方法难以评估的治疗表达变化细微的疾病可能有益。肌肉神经 58: 286-292, 2018.
