Subcellular analysis of Tyr-aminopeptidase activities in the developing rat cerebellum

The endogenous opioid system seems to play important roles in the developing cerebellum. The first opioid peptide isolated, Met-enkephalin, is expressed transiently in this brain area. In the present study, several enzyme activities capable of hydrolyzing enkephalins are measured during the first month of cerebellar development, using Tyr-beta-naphthylamyde as substrate and puromycin as inhibitor of one of the membrane-bound aminopeptidases. Puromycin-sensitive soluble and membrane-bound aminopeptidase activities decrease in the synaptosomal and mitochondrial fractions at the end of the first month of life, just when enkephalin-like immunoreactivity decreases in the cerebellum. Membrane-bound enzyme also decreases in the myelinic fraction. Synaptosomal activity increases after birth, coinciding with decreases in the activity in the microsomal fraction. Puromycin-insensitive and membrane-bound aminopeptidase shows less significant developmental changes and they occur mainly in the first week of life, coinciding with the axonal and dendrite growth. These results could suggest a possible role of these enzymes, together with the rest of the opioid system, in cerebellar development.