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Cytosine Arabinoside Induces Apoptosis in Cerebellar Neurons in Culture
Author(s) -
Dessi Frédéric,
Pollard Hélène,
Moreau Joelle,
BenAri Yezekiel,
CharriautMarlangue Christiane
Publication year - 1995
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1046/j.1471-4159.1995.64051980.x
Subject(s) - dna fragmentation , microbiology and biotechnology , biology , apoptosis , agarose gel electrophoresis , cytosine , fragmentation (computing) , dna , programmed cell death , gel electrophoresis , cell culture , dna synthesis , rna , biochemistry , gene , genetics , ecology
Cytosine arabinoside (AraC) is a pyrimidine antimetabolite that prevents cell proliferation by inhibiting DNA synthesis. We report that AraC kills cultured cerebellar neurons in a concentration‐dependent fashion with an EC 50 of ∼60 µ M when added shortly after seeding. This cell death has apoptotic features because we observed (1) morphology of apoptotic nuclei as judged by DNA staining with Hoechst 33258, (2) DNA fragmentation with typical ladder pattern on agarose gel, (3) positive nuclear labeling with a specific in situ DNA fragmentation staining, (4) prevention by deoxycytidine (IC 50 = 1 µ M ), protein, and RNA synthesis inhibitors, and (5) release of DNA fragments in the incubating medium. We have also observed that several proteins were overexpressed in AraC‐treated neurons by two‐dimensional polyacrylamide gel electrophoresis. We conclude that AraC induces a signal that triggers a cascade of new mRNA and protein synthesis, leading to apoptotic cell death in cultured cerebellar granule cells.