Alterations in the Multivariate Organization of Plasma Fatty Acid Profiles and Spontaneous Behavior in an...

Current experimental neurotoxicity research commonly relies on evaluating individual behavioral and biochemical endpoints, often without considering whether spontaneous behavior and lipid metabolism exhibit coordinated multivariate organization under toxicological stress. To address this question, the present study employed an integrative framework combining spontaneous behavioral profiling, targeted plasma fatty acid methyl ester (FAME) profiling, chemometric analyses, targeted assessment of cholinergic function, and permutation-based statistical validation in an AlCl₃-induced rat model of neurotoxicity. Behavioral assessment was performed using the Y-maze, and multivariate organization was evaluated using covariance analyses, principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), one-dimensional projection modeling, and nonparametric permutation testing (10,000 permutations). AlCl₃ exposure was associated with early dose-related behavioral impairment at day 21 (ANOVA, p = 0.0006). In addition, both the measured fatty acid profiles and behavioral measures exhibited significant non-random multivariate organization, with progressive dose-related changes observed following AlCl₃ exposure. Collectively, these findings suggest that neurotoxic exposure is associated with coordinated changes in the multivariate organization of measured fatty acid profiles, behavioral measures, and cholinergic function rather than isolated alterations in individual variables. Although confirmation in larger independent cohorts and broader molecular profiling remains warranted, the present findings support the use of multivariate organizational analysis as a complementary systems-level framework for investigating experimental neurotoxicity.