Cytoplasmic accumulations in rat primary brain cell cultures following treatment with E-64, a thiol protease inhibitor.
The present communication deals with morphological effects of E-64, a thiol protease inhibitor, on primary cultures dissociated from 15-day-old fetal rat brain. Brain cells were treated with different concentrations of E-64 varying from 0.1 to 50 micrograms/ml medium. Numerous cytoplasmic accumulations appeared in neuronal cells, astrocytes and oligodendrocytes exposed to a concentration of 1.0 micrograms/ml of E-64 or higher, and after withdrawal of E-64, the accumulations disappeared. These accumulations were immunocytochemically stained with the antibody against cathepsin B, a marker of lysosomal activity, and the staining patterns found with acid phosphatase were similar to those found with cathepsin B. It is hypothesized that the cytoplasmic accumulations were lysosomes and that the action of E-64 was reversible. Use of thiol protease inhibitors on primary brain cell culture may be an appropriate model system for studying protein storage diseases in neuronal cells.
Matsui
K
K
Department of Pediatrics, Jikei University School of Medicine, Tokyo, Japan.
Shirasawa
N
N
Eto
Y
Y
D000818:Animals; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000648:ultrastructure*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D007930:Leucine / Q000031:analogs & derivatives* / Q000494:pharmacology; D008578:Meninges / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008854:Microscopy, Electron; D051381:Rats; D011919:Rats, Inbred Strains
D004791:Enzyme Inhibitors; D003546:Cysteine Endopeptidases; D007930:Leucine; C024974:E 64