Biochemical investigation to the reliability of the histochemical demonstration of microsomal arylsulphatase activity in cryostat sections.
Polyacrylamide gel-electrophoresis was performed with an extract from cultivated skin fibroblasts. Arylsulphatase activity is measured and visualised using the biochemical substrate dehydroepiandrosterone sulphate and the histochemical substrate 6-bromo-2-naphthyl sulphate respectively. The histochemical substrate was hydrolysed at Rf = 0.49 and 0.58 while the biochemical substrate was hydrolysed only at 0.49. We conclude that two different microsomal arylsulphatases exist: a sulphatase able to hydrolyse steroid sulphatases (Rf = 0.49) and one unable to hydrolyse steroid sulphatases (RF = 0.58). In consequence if is recommended to carry out an electrophoresis experiment after the histochemical investigation, in order to discriminate between these two types of sulphatase.
D001192:Arylsulfatases / Q000302:isolation & purification*; D003687:Dehydroepiandrosterone; D005347:Fibroblasts / Q000201:enzymology; D006651:Histocytochemistry; D006801:Humans; D008861:Microsomes / Q000201:enzymology*; D009281:Naphthalenes; D012867:Skin / Q000201:enzymology; D013429:Sulfatases / Q000302:isolation & purification*
C021156:6-bromonaphthyl sulfate; D009281:Naphthalenes; D003687:Dehydroepiandrosterone; D013429:Sulfatases; D001192:Arylsulfatases