Binding and internalization of lysosomal enzymes by primary cultures of rat glia.
Highly purified cultures of rat astrocytes and oligodendrocytes were examined for their ability to bind and internalize lysosomal enzymes. Astrocytes displayed a saturable uptake of beta-glucosidase and beta-galactosidase. The uptake was specifically inhibited by mannose-6-phosphate but not by several other sugars or sugar phosphates, indicating that the process was mediated by mannose-6-phosphate receptors. When cells were allowed to take up 125I-beta-glucosidase for 1 hr at 37 degrees C and subcellular organelles were isolated, the enzyme was shown to comigrate with a lysosomal organelle marker enzyme, suggesting that the enzyme was targeted to lysosomes. Astrocyte receptors were probed directly by binding of 125I labeled beta-glucosidase to astrocyte membranes at 4 degrees C. Binding was saturable and competitively inhibited by mannose-6-phosphate. In contrast to the astrocytes, cultured oligodendrocytes showed no specific binding or uptake of the lysosomal enzymes. Immunocytochemical staining of mixed glial cultures supported the biochemical data; only the astrocytes stained positive with anti-mannose-6-phosphate receptor antibodies.
Hill
D F
DF
Bullock
P N
PN
Chiappelli
F
F
Rome
L H
LH
D000818:Animals; D000831:Animals, Newborn; D001253:Astrocytes / Q000201:enzymology*; D002352:Carrier Proteins / Q000502:physiology; D002463:Cell Membrane Permeability; D002478:Cells, Cultured; D002540:Cerebral Cortex; D005696:Galactosidases / Q000378:metabolism*; D005959:Glucosidases / Q000378:metabolism*; D008247:Lysosomes / Q000201:enzymology*; D009457:Neuroglia / Q000201:enzymology*; D009836:Oligodendroglia / Q000201:enzymology*; D051381:Rats; D011919:Rats, Inbred Strains; D017527:Receptor, IGF Type 2; D001616:beta-Galactosidase / Q000378:metabolism*; D001617:beta-Glucosidase / Q000378:metabolism*
D002352:Carrier Proteins; D017527:Receptor, IGF Type 2; D005696:Galactosidases; D005959:Glucosidases; D001617:beta-Glucosidase; D001616:beta-Galactosidase