5-Bromo-2'-deoxyuridine is selectively toxic to neuronal precursors in vitro.
The effect of 5-bromo-2'-deoxyuridine (BrdU) incorporation on the phenotype of progeny derived from expanded E18 rat striatal precursors was examined. BrdU was administered to cultures for 24 h prior to differentiation. Results revealed that there was selective toxicity of this compound to developing TuJ1+ neurons, but not glia, at concentrations used in most labelling studies. Therefore, a BrdU dose-response curve from 0.2 microM to 10 microM was established. The optimum dose of BrdU for labelling cells was 0.2 microM, well below the 1-10 microm recommended concentration. This dose resulted in the survival of significantly more newborn BrdU/TuJ1+ double-labelled neurons and eliminated the toxic effects of BrdU. Administration of 10 microm BrdU resulted in a significant decrease in extracellular regulated kinase (ERK) phosphorylation compared with untreated cultures, this could be completely restored by the administration of either N-methyl-D-aspartate (NMDA) receptor antagonists such as MK801 or the nitric oxide synthesis inhibitor L-methyl-arginine methyl ester (L-NAME). Our results show that high levels of BrdU are selectively toxic to neurons through a mechanism that activates classical cell death pathways. This has implications for labelling studies both in vivo and in vitro.
Caldwell
Maeve A
MA
Centre for Brain Repair and Department of Clinical Neurosciences, Cambridge University Forvie Site, Robinson Way, Cambridge CB2 2PY, UK. mac28@hermes.cam.ac.uk
He
Xiaoling
X
Svendsen
Clive N
CN
D000818:Animals; D000963:Antimetabolites / Q000633:toxicity*; D001253:Astrocytes / Q000187:drug effects / Q000378:metabolism; D015153:Blotting, Western; D001973:Bromodeoxyuridine / Q000633:toxicity*; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D016291:Dizocilpine Maleate / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D018691:Excitatory Amino Acid Antagonists / Q000494:pharmacology; D005904:Glial Fibrillary Acidic Protein / Q000378:metabolism; D008938:Mitosis / Q000187:drug effects; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects*; D052248:Nitric Oxide Synthase Type I / Q000037:antagonists & inhibitors; D009836:Oligodendroglia / Q000187:drug effects / Q000378:metabolism; D010766:Phosphorylation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013234:Stem Cells / Q000187:drug effects*
D000963:Antimetabolites; D004791:Enzyme Inhibitors; D018691:Excitatory Amino Acid Antagonists; D005904:Glial Fibrillary Acidic Protein; D016291:Dizocilpine Maleate; D052248:Nitric Oxide Synthase Type I; D001973:Bromodeoxyuridine; D019331:NG-Nitroarginine Methyl Ester