Diagnosis of arylsulfatase A deficiency in intact cultured cells using a fluorescent derivative of cerebroside sulfate.
A fluorescent derivative of cerebroside sulfate (12-(1-pyrene)dodecanoyl-sphingosylgalactosyl-0-3-sulfate (P12-sulfatide) has been synthesized as a potential substrate for the determination of cerebroside sulfatidase (or arylsulfatase A) activity. It was administered into cultured human skin fibroblasts and thereby utilized for the diagnosis of arylsulfatase A deficiency. Cultured skin fibroblasts from normal individuals and healthy persons suffering from a pseudoarylsulfatase A deficiency (PD) degraded the P12-sulfatide, while in cells derived from a metachromatic leukodystrophy (MLD) patient it remained essentially intact. This contrasts with in vitro determinations of enzymatic activity, where the MLD or PD-derived arylsulfatase A exhibit similar deficiency, in spite of a profoundly different clinical course. Administration of the fluorescent sulfatide into the intact cells permitted a sensitive and rapid diagnosis of MLD and its distinction from the PD-phenomenon. This might be of particular importance for cases in which a rapid diagnosis is required and for prenatal diagnosis of fetuses from families afflicted with both MLD and pseudo-deficiency mutant genes.
D002460:Cell Line; D002553:Cerebroside-Sulfatase / Q000172:deficiency*; D005260:Female; D005456:Fluorescent Dyes; D006801:Humans; D007966:Leukodystrophy, Metachromatic / Q000175:diagnosis* / Q000201:enzymology / Q000235:genetics; D011247:Pregnancy; D011296:Prenatal Diagnosis; D013433:Sulfoglycosphingolipids
D005456:Fluorescent Dyes; D013433:Sulfoglycosphingolipids; C052759:12-(1-pyrenedodecanoyl)sphingosylgalactosyl-O-3-sulfate; D002553:Cerebroside-Sulfatase