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2′‐Deoxyadenosine Induces Apoptosis in Rat Chromaffin Cells
Author(s) -
Wakade Arun R.,
Guo Xi,
Palmer Kenneth C.,
Kulkarni Jayant S.,
Przywara Dennis A.,
Wakade Taruna D.
Publication year - 1996
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.1471-4159.1996.67062273.x
Subject(s) - deoxyadenosine , biology , programmed cell death , apoptosis , microbiology and biotechnology , deoxycoformycin , dna laddering , nucleoside , biochemistry , adenosine , dna fragmentation , adenosine deaminase
We show here that 2′‐deoxyadenosine (2′‐dAdo) but not adenosine was toxic to chromaffin cells of 3–4‐week‐old rat adrenal glands. More than 75% of the cells plated in culture gradually died over a 3‐day period in the presence of 100 µ M 2′‐dAdo plus 3 µ M deoxycoformycin (DCF). Morphological observations together with bisbenzimide staining and terminal deoxynucleotidyl transferase‐mediated nick end labeling showed membrane blebbing, shrinkage of cell bodies, chromatin condensation, and DNA fragmentation, suggesting apoptosis‐like cell death by 2′‐dAdo. Lethal effects of 2′‐dAdo were potentiated by DCF, a drug that inhibits adenosine deaminase. 2′‐dAdo‐prompted cell death was not prevented by inhibitors of nucleoside transporter (3 µ M dilazep or 1 µ M nitrobenzylthioinosine), precursors of pyrimidine nucleotide biosynthesis (300 µ M uridine or 100 µ M 2′‐deoxycytidine), or 5 m M nicotinamide. Cells incubated with 2′‐dAdo (100 and 300 µ M ) showed a three‐ and ninefold, respectively, increase in content of dATP, a product known to be an inhibitor of ribonucleotide reductase, an enzyme essential for DNA synthesis. Formation of dATP was completely prevented by iodotubercidin (ITu), a drug that inhibits phosphorylation of 2′‐dAdo to dATP by nucleoside kinase. It is interesting that nanomolar concentrations of ITu also completely protected chromaffin cells from 2′‐dAdo lethality. Our study demonstrates for the first time that mammalian adrenal chromaffin cells undergo apoptotic cell death by a natural nucleoside and suggests that this model could be used to study apoptosis and cell function.