Liver protein expression in F1 offspring arising from spermatogonia exposed to 60 cGy of fission spectrum neutrons from the JANUS reactor was compared to that in offspring from unexposed spermatogonia by using two-dimensional electrophoresis (2DE). Approximately 100 protein spots in 2DE patterns from 167 control offspring and 530 offspring from irradiated sires were monitored for quantitative decreases of 50%, indicative of mutation events causing the loss of one normal copy of a structural gene. Reproducible abnormalities were found only in 3 patterns, all from the offspring of neutron-irradiated sires. Two of the three patterns were from littermates (brother and sister) and both showed an approximately 70% decrease in the amount of liver protein MSN188. The third pattern was from a male mouse sired by a different male and showed an approximately 50% decrease in the abundance of protein MSN94. The decreased abundance of MSN188 and MSN94 was assumed to be due to mutation events referred to as NEUT1 and NEUT2, respectively. Sibling crosses between the 2 mice showing the NEUT1 trait produced offspring with control, decreased and undetectable levels of MSN188 in a ratio of 0.25:0.5:0.25. Test crosses between the F1 offspring expressing the NEUT2 trait back to C57BL/6JANL mice produced offspring expressing normal or decreased amounts of MSN94 in a ratio of 0.5:0.5. Inbreeding of individuals expressing decreased amounts of MSN94 produced mice expressing control, decreased amounts, or no detectable amount of that protein in a ratio of 0.25:0.5:0.25. These results indicate that the decreased abundance of MSN188 or MSN94 originally detected in the F1 offspring is due to a genetically transmissible event. Unlike the heritable protein changes observed previously in the F1 offspring of sires exposed to N-ethyl-N-nitrosourea in which a protein variant was produced, both the NEUT1 and NEUT2 mutation events appear to prevent the production of any protein product. These 2 mutations may thus represent mutation lesions other than point mutations (e.g., deletions or translocations) detectable as quantitative changes in protein expression in the F1 generation.