N-(1,1-dimethyl-3-oxobutyl)acrylamide

Neurotoxicity

Administrative data

Endpoint:

neurotoxicity

Remarks:

other: in vitro

Type of information:

experimental study

Adequacy of study:

supporting study

Study period:

1990

Reliability:

2 (reliable with restrictions)

Rationale for reliability incl. deficiencies:

other: Publication in a recognized journal.

Data source

Reference

Reference Type:

publication

Title:

Unnamed

Year:

1991

Materials and methods

Test guideline

Qualifier:

no guideline followed

Principles of method if other than guideline:

Dorsal root ganglia (DRG) from 3-day-old rats were used. DRG was evaluated in culture as a screening system by studying the relation of the neurotoxicity of test chemicals in this culture to their acute toxicity and rate constant with glutathione. Acrylamide and 6 derivatives were investigated.

GLP compliance:

not specified

Limit test:

no

Test material

Test material information

Constituent 1

Reference substance name:

diacetone acrylamide

IUPAC Name:

diacetone acrylamide

Details on test material:

Supplier: Tokyo Kasei Co.

Test animals

Species:

other: In vitro: dorsal root ganglia (DRG) from 3-day-old rats.

Administration / exposure

Details on study design:

The dorsal root ganglia from 3-day-old rats were taken out under a dissecting microscope and collected in a plastic dish containing phosphate-buffered saline (PBS). These ganglia were then incubated for 30 min in PBS containing 0.05 % trypsin at 37 °C, followed by washing with PBS. One ganglion explant was placed in a dish coated with collagen and cultured in Dulbecco's Modified Eagle Medium (DMEM, Gibco). The medium was supplemented with 10 % fetal calf serum (Gibco), penicillin (100 units/ml, Gibco), Streptomycin (100 units/ml, Gibco), 25 µM cytosine arabinofuranoside and 2.5S nerve growth factor (10 ng/ml) in a final volume of 1.0 ml at 37 °C in a humidified atmosphere of 10 % C02. Test chemicals were added at various concentrations to the medium containing the explant at the beginning of culture. After culturing for 4 days, the neurite growth from the explant was observed under a phase-contrast microscope, and photographed on Fuji NEOPAN F films. To quantify the inhibitory effects of test compounds, a scoring of the neurite growth was done. The half-maximum inhibition concentration (I50) was determined from the dose-response curve.

Results and discussion

Any other information on results incl. tables

Acrylamide and 6 derivatives inhibited neurite growth from rat dorsal root ganglion in culture. The half-maximum inhibition concentration varied among test compounds, ranging from 0.8 mM for acrylamide and N-hydroxymethylacrylamide to 30 mM for methacrylamide. The value correlated well with the acute oral LD50 for rats.

For the test chemicals, the neurotoxicity obtained in this culture system did not correlate with the neurotoxicity in vivo.

Applicant's summary and conclusion

Executive summary:

Acrylamide and 6 derivatives (including diacetone acrylamide) inhibited neurite growth from rat dorsal root ganglion in culture. The half-maximum inhibition concentration varied among test compounds, ranging from 0.8 mM for acrylamide and N-hydroxymethylacrylamide to 30 mM for methacrylamide. The value correlated well with the acute oral LD50 for rats.

For the test chemicals, the neurotoxicity obtained in this culture system did not correlate with the neurotoxicity in vivo.