Screening for developmental neurotoxicity using PC12 cells: comparisons of organophosphates with a carbamate, an organochlorine, and divalent nickel.
BACKGROUND
In light of the large number of chemicals that are potential developmental neurotoxicants, there is a need to develop rapid screening techniques.


OBJECTIVE
We exposed undifferentiated and differentiating neuronotypic PC12 cells to different organophosphates (chlorpyrifos, diazinon, parathion), a carbamate (physostigmine), an organochlorine (dieldrin), and a metal (divalent nickel; Ni2+) and examined indices of cell replication and differentiation for both short- and long-term exposures.


RESULTS
In undifferentiated cells, all the agents inhibited DNA synthesis, with the greatest effect for diazinon, but physostigmine eventually produced the largest deficits in the total number of cells after prolonged exposure. The onset of differentiation intensified the adverse effects on DNA synthesis and changed the rank order in keeping with a shift away from noncholinergic mechanisms and toward cholinergic mechanisms. Differentiation also worsened the effects of each agent on cell number after prolonged exposure, whereas cell growth was not suppressed, nor were there any effects on viability as assessed with trypan blue. Nevertheless, differentiating cells displayed signs of oxidative stress from all of the test compounds except Ni2+, as evidenced by measurements of lipid peroxidation. Finally, all of the toxicants shifted the transmitter fate of the cells away from the cholinergic phenotype and toward the catecholaminergic phenotype.


CONCLUSIONS
These studies point out the feasibility of developing cell-based screening methods that enable the detection of multiple end points that may relate to mechanisms associated with developmental neurotoxicity, revealing some common targets for disparate agents.
Slotkin
Theodore A
TA
Department of Pharmacology & Cancer Biology, Duke University Medical Center, Durham, North Carolina 27710, USA. t.slotkin@duke.edu
MacKillop
Emiko A
EA
Ryde
Ian T
IT
Tate
Charlotte A
CA
Seidler
Frederic J
FJ
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002795:Choline O-Acetyltransferase / Q000378:metabolism; D002800:Cholinesterase Inhibitors / Q000633:toxicity; D004247:DNA / Q000096:biosynthesis; D004026:Dieldrin / Q000633:toxicity*; D007306:Insecticides / Q000633:toxicity; D009532:Nickel / Q000633:toxicity*; D009943:Organophosphorus Compounds / Q000633:toxicity*; D016716:PC12 Cells; D010830:Physostigmine / Q000633:toxicity*; D051381:Rats; D018675:Toxicity Tests / Q000379:methods*
D002800:Cholinesterase Inhibitors; D007306:Insecticides; D009943:Organophosphorus Compounds; D009532:Nickel; D004247:DNA; D010830:Physostigmine; D002795:Choline O-Acetyltransferase; D004026:Dieldrin