Gravel R A, Leung A
Hum Genet. 1983;65(2):112-6. doi: 10.1007/BF00286645.
Gaucher disease is a lysosomal storage disorder resulting from a deficiency of acid beta-glucosidase. Several clinical forms have been described, including infantile, juvenile, and adult onset variant. We have examined complementation in infantile and adult forms of Gaucher disease by monitoring enzyme activity in multinucleate cells produced by fusing skin fibroblasts from different patients in the presence of polyethylene glycol. beta-Glucosidase activity was monitored in lysates of individual multinucleate cells by a microassay method utilizing methylumbelliferyl-beta-D-glucoside as the substrate (normal: 1.3 +/- 0.12 x 10(-13) mol/h/cell). The microassay was linear with time up to 4 h, for up to 20 mononucleate cells, and for individual multinucleate cells containing up to 12 nuclei. Complementation was examined in 11 fibroblasts strains fused in all pairwise combinations. In no instance was there any clear indication of complementation (at least 10-15% of normal activity to adequately account for experimental variation) although there was an indication of marginal increases in some fusions. On the other hand, the expected 50% activity was obtained in "heterozygous" fusions (normal/mutant) for both types of clinical variants. Our results are consistent with a single gene, presumably the structural gene encoding the enzyme, responsible for at least the infantile and adult variants, and confirm the autosomal recessive nature of the disorder.
戈谢病是一种由于酸性β-葡萄糖苷酶缺乏导致的溶酶体贮积症。已描述了几种临床类型，包括婴儿型、青少年型和成人型。我们通过在聚乙二醇存在下融合来自不同患者的皮肤成纤维细胞产生的多核细胞中监测酶活性，研究了婴儿型和成人型戈谢病的互补作用。利用甲基伞形酮基-β-D-葡萄糖苷作为底物，通过微量测定法监测单个多核细胞裂解物中的β-葡萄糖苷酶活性（正常：1.3±0.12×10⁻¹³摩尔/小时/细胞）。该微量测定法在长达4小时内、对于多达20个单核细胞以及对于含有多达12个核的单个多核细胞而言，活性与时间呈线性关系。对11个成纤维细胞株进行了所有两两组合的融合，并检测了互补作用。在任何情况下都没有明显的互补迹象（至少要有正常活性的10 - 15%才能充分解释实验差异），尽管在某些融合中显示有少量增加。另一方面，对于两种临床变体，在“杂合”融合（正常/突变）中均获得了预期的50%活性。我们的结果与单个基因一致，推测该基因是编码该酶的结构基因，至少负责婴儿型和成人型变体，并证实了该疾病的常染色体隐性性质。
