Kopitz J, Gerhard C, Höfler P, Cantz M
Institute of Pathochemistry and General Neurochemistry, University of Heidelberg, Germany.
Clin Chim Acta. 1994 Jun;227(1-2):121-33. doi: 10.1016/0009-8981(94)90141-4.
Incorporation of the lysosomotropic amine [14C]methylamine by fibroblasts cultured from patients with lysosomal storage diseases and from controls was used to estimate the size of the lysosomal compartment. All cell lines from patients with infantile and juvenile forms of mucopolysaccharidoses, mucolipidoses and oligosacharidoses showed markedly increased radioactivity compared with the normal range of controls. In cells from patients with sphingolipidoses and adult forms of storage diseases, however, methylamine accumulation was not significantly increased. Experimentally induced lysosomal storage by enzyme inhibitors (leupeptin, suramin) also caused increased methylamine accumulation. When the lysosomal pH was determined with fluorescein isothiocyanate-dextran, it was in the range of normal controls (pH 4.7-5.0) in patients cells. Thus, [14C]methylamine accumulation should depend on the volume rather than differences in acidity of the lysosomal compartment and be a measure of its eventual pathological enlargement. We conclude that the determination of [14C]methylamine accumulation in fibroblasts provides a valuable tool in the screening for a variety of lysosomal storage disorders.
利用溶酶体亲和胺[¹⁴C]甲胺被溶酶体贮积病患者和对照者培养的成纤维细胞摄取的情况来估计溶酶体区室的大小。与正常对照范围相比,所有来自婴儿型和青少年型黏多糖贮积症、黏脂贮积症和寡糖贮积症患者的细胞系都显示出明显增加的放射性。然而,在来自鞘脂贮积症患者和成人型贮积病患者的细胞中,甲胺积累没有显著增加。用酶抑制剂(亮抑酶肽、苏拉明)实验性诱导的溶酶体贮积也导致甲胺积累增加。当用异硫氰酸荧光素 - 葡聚糖测定溶酶体pH时,患者细胞的pH在正常对照范围内(pH 4.7 - 5.0)。因此,[¹⁴C]甲胺积累应取决于溶酶体区室的体积而非酸度差异,并且是其最终病理性增大的一个指标。我们得出结论,测定成纤维细胞中[¹⁴C]甲胺积累为筛查多种溶酶体贮积症提供了一种有价值的工具。