Research activities within the Molecular Neurotoxicology Laboratory are directed toward the discovery and characterization of gene-activation and cell-signaling pathways associated with neural injury induced gliosis. A related focus is to apply our findings concerning the molecular basis of neurotoxicity to examine the influence of susceptibility factors (obesity, age, stress and gender) on neurotoxic outcomes. Technically, the lab focuses on the use of in vivo and ex vivo models using conventional rats and mice as well as transgenic “knock-ins” and “knock-outs.” Ex vivo and in vitro work takes advantage of the brain-slice preparation. Methodology has been developed or established to examine the common-down stream effectors in the dopamine/ hormone/stress/inflammation/trophic factor cascades: the PKA, MAP, SAP, CDK, and JAK-STAT kinase phosphatase/phosphorylation modules. Specifically, phospho-state specific antibodies are being utilized to examine the in vivo status of specific phosphorylation pathways. Coupled with use of antibodies already available and with our demonstration of a technique to “fix” phosphorylation state, in vivo (Focused high intensity microwave fixation), it is possible to identify altered phosphorylation states associated with drug and toxicant-induced effects on the basal ganglia or other brain regions of interest. Phosphorylation studies are being complimented by examination of gene expression events (e.g. fos-related antigens, heat-shock proteins, glial genes linked to astrocyte hypertrophy and microglial activation) associated with altered signaling using the Affymetrix gene-array platform. Traditional RT-PCR, filter expression arrays (CloneTech) and real-time quantitative PCR (Taqman) are routinely employed in our studies. The results of these expression studies, in turn, are carried out in conjunction with assays for an array of neuronal and glial proteins using robotically-processed ELISAs developed in the Molecular Neurotoxicology Laboratory over the last decade. In addition, our recent acquisition of SELDI technology has allowed us to pursue more open-ended discovery studies aimed at developing novel biomarkers of neurotoxicity as well as more broad-scale screening of protein phosphorylation modules affected by a variety of neural injury models. Finally, in addition to outside collaborators, extensive neuroanatomical support is available in-house through collaborators and a morphology core containing a confocal microscope and an unbiased stereology morphometry system. Ultimately, our goal is to identify the earliest molecular signatures emanating from any damaged cell in the nervous system to intervene therapeutically to ameliorate or prevent irreversible damage to the developing or adult central nervous system.