CEREBROSIDE GALACTOSIDASE: A METHOD FOR DETERMINATION AND A COMPARISON WITH OTHER LYSOSOMAL ENZYMES IN DEVELOPING RAT BRAIN1

Abstract- ( 1) A method is described for assaying brain for cerebroside galactosidase activity. The enzyme was liberated by sonication and addition of sodium taurocholate, then by digestion with pancreatic enzymes. It was further purified by precipitation at pH 3. The enzyme was then incubated with an emulsion of galactose-labelled cerebroside in tauro-cholate and oleate at pH 4.5, and the liberated galactose was determined by scintillation counting. (2) The content of cerebroside galactosidase in rat brain at various ages has been determined. The enzyme was present before cerebroside appears in noticeable amounts (4 days) and the amount rose considerably during the period of active cerebroside deposition and myelination. The amount then remained at a high concentration even in the adult. (3) Comparison with other lysosomal brain enzymes was made in the age study. Nitro-phenyl galactoside hydrolase also increased during myelination but levelled off earlier; its activity paralleled the amount of ganglioside. Nitrophenyl glucoside hydrolase started at a lower level and decreased with age. Sulphatase activity rose during myelination, then decreased somewhat after 15 days. Ceramidase followed a pattern similar to that of nitro