3 RS APPROACHES FOR THE STUDY OF SIGNALING EVENTS TRIGGERED BY PESTICIDE INDUCED NEUROTOXICITY.

CONDE, MELISA A.; MANISCALCHI, ATHINA; NICASIO, MARÍA L.; BENZI JUNCOS, ORIANA N.; FUNK, MELANIA I.; ALZA, NATALIA P.; SALVADOR, GABRIELA A.

Congreso; REUNIÓN CONJUNTA SAIC SAB AAFE AACYTAL 2023; 2023

Maneb (MB), a dithiocarbamate pesticide, can cross the blood-brainbarrier and is considered an environmental risk factor associatedwith Parkinsonism. Our aim was to characterize redox signalingduring MB-induced neurotoxicity in different experimental modelsaccording to the 3Rs principle.Taking into account full Replacement we firstly exposed neuronalcell line cultures to MB and we detected increased expression levelsof the redox sensitive transcription factor Nrf2 and its regulator Sirt1.Neuronal MB exposure also triggered the increase of lipid peroxidation,GSH depletion, GpX4 downregulation and mitochondrial alterations,indicating that ferroptosis is involved in the mechanism of celldeath. We confirmed these results in a model of relative Replacementby using glial mixed primary cultures. We also demonstratethat Nrf2/Sirt1 signaling is a protective strategy against neurotoxicity.Based on these results and with the aim of evaluating phenotypicalaspects for establishing a preclinical model, we design an experimentalparadigm with C57BL/6 mice. Following Refinement criteria,treatments were performed with the supervision of a veterinarianwho care the animal welfare during the entire procedure. C57BL/6mice (18-20g) were intraperitoneally injected with MB (100 mg/kg)for 6 weeks. Behavioral tests (Open Field and Rotarod) were performedon week 3 and 6 of treatment. One day after the last administration,animals were sacrificed by cervical dislocation. Forebrain, midbrain and hindbrain were separated for molecular determinations.To apply the Reduction criteria all other animal organs wereused for related investigations. MB-treated animals showed a decreasein total distance travelled, supported and unsupported rearsand in falling latency. These results indicate that MB neurotoxicitytriggers a parkinsonian phenotype in C57BL/6.The results presented here constitute a platform for toxicologicaland preclinical studies according to 3Rs principles.