QUANTIFICATION OF LECTIN FLUORESCENCE IN GNE MYOPATHY MUSCLE BIOPSIES
QUANTIFICATION OF LECTIN FLUORESCENCE IN GNE MYOPATHY MUSCLE BIOPSIES
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DOI：
10.1002/mus.26135
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发表时间：
2018-08-01
期刊：
影响因子：
3.4
通讯作者：
Malicdan, May Christine
中科院分区：
文献类型：
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作者：
Leoyklang, Petcharat;Class, Bradley;Malicdan, May Christine
GNE肌病是一种成人发病的肌肉疾病，其特征是（肌肉）聚糖的唾液化受损，可通过凝集素组织化学检测。我们描述了一种标准化的方法来量化（凝集素-）荧光在肌肉切片，适用于诊断和治疗反应的GNE肌病。方法：用唾液酸结合的黑参凝集素（SNA）和肌膜停留蛋白CAV-3抗体对肌肉切片进行荧光标记。整个组织切片在瓦片中成像并定量荧光。结果：与肌肉匹配的对照组相比，与CAV-3共定位的SNA荧光在GNE肌病活检中减少了50%，证实了先前的定性结果。讨论：该定量荧光法可准确测定GNE肌病肌肉活检组织唾液化状态。该方法适用于其他膜相关肌肉蛋白的表达，并且可能对表达治疗变化微妙且难以通过其他方法评估的疾病有益。
Introduction: 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. Methods: 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. Results: SNA fluorescence co-localizing with CAV-3 was similar to 50% decreased in GNE myopathy biopsies compared with muscle-matched controls, confirming previous qualitative results. Discussion: 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.