Acrylamide,2,5-Hexanedione and β-Aminopropionitrile Toxicity Tested in Rat Embryo Mid-brain Cell Cultures

Abstract

The abilities of three substances, acrylamide, 2,5-hexanedione and β-aminopropionitrile to affect nerve cell differentiation were compared with their general cytotoxicities (ability to affect cell survival) over five days in rat embryo mid-brain micromass cultures. ACA and 25HD are neurotoxins in the adult, but only ACA is known to have transplacental effects on the developing central nervous system. BAPN is a model teratogen, but not a neurotoxin, in the adult and was used as a positive control. ACA inhibited neuronal differentiation (assayed by reduction of number of foci of differentiated neurons) by 50% at a concentration (the IC50) of 15μg/ml. The corresponding IC50 for cell survival (assayed by reduction in cell number) was 36μg/ml. In 25HD-exposed cultures, IC50 values of 729 and 872μg/ml were obtained for the inhibition of differentiation and cell survival, respectively. BAPN inhibited differentiation with an IC50 of 226μg/ml, but had very little effect on cell viability up to 500μg/ml. This pronounced difference in sensitivity of the two endpoints is in agreement with previous reports of its teratogenicity. The more-limited but significant difference detected for ACA, but not for 25HD, suggests that rat embryo mid-brain micromass cultures can be used to study subteratogenic effects of ACA on brain development.

Keywords

acrylamide 2,5-hexanedione β-aminopropionitrile in vitro embryotoxicity midbrain cell cultures

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