PMID- 34513802 OWN - NLM STAT- PubMed-not-MEDLINE LR - 20220514 IS - 2296-4185 (Print) IS - 2296-4185 (Electronic) IS - 2296-4185 (Linking) VI - 9 DP - 2021 TI - Chlorpyrifos Disrupts Acetylcholine Metabolism Across Model Blood-Brain Barrier. PG - 622175 LID - 10.3389/fbioe.2021.622175 [doi] LID - 622175 AB - Despite the significant progress in both scientific understanding and regulations, the safety of agricultural pesticides continues to be called into question. The need for complementary analytics to identify dysregulation events associated with chemical exposure and leverage this information to predict biological responses remains. Here, we present a platform that combines a model organ-on-chip neurovascular unit (NVU) with targeted mass spectrometry (MS) and electrochemical analysis to assess the impact of organophosphate (OP) exposure on blood-brain barrier (BBB) function. Using the NVU to simulate exposure, an escalating dose of the organophosphate chlorpyrifos (CPF) was administered. With up to 10 muM, neither CPF nor its metabolites were detected across the BBB (limit of quantitation 0.1 microM). At 30 microM CPF and above, targeted MS detected the main urinary metabolite, trichloropyridinol (TCP), across the BBB (0.025 microM) and no other metabolites. In the vascular chamber where CPF was directly applied, two primary metabolites of CPF, TCP and diethylthiophosphate (DETP), were both detected (0.1-5.7 microM). In a second experiment, a constant dose of 10 microM CPF was administered to the NVU, and though neither CPF nor its metabolites were detected across the BBB after 24 h, electrochemical analysis detected increases in acetylcholine levels on both sides of the BBB (up to 24.8 +/- 3.4 microM) and these levels remained high over the course of treatment. In the vascular chamber where CPF was directly applied, only TCP was detected (ranging from 0.06 muM at 2 h to 0.19 muM at 24 h). These results provide chemical evidence of the substantial disruption induced by this widely used commercial pesticide. This work reinforces previously observed OP metabolism and mechanisms of impact, validates the use of the NVU for OP toxicology testing, and provides a model platform for analyzing these organotypic systems. CI - Copyright (c) 2021 Miller, McClain, Dodds, Balinski, May, McLean and Cliffel. FAU - Miller, Dusty R AU - Miller DR AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. FAU - McClain, Ethan S AU - McClain ES AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. FAU - Dodds, James N AU - Dodds JN AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. AD - Center for Innovative Technology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, United States. FAU - Balinski, Andrzej AU - Balinski A AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. AD - Center for Innovative Technology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, United States. FAU - May, Jody C AU - May JC AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. AD - Center for Innovative Technology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, United States. FAU - McLean, John A AU - McLean JA AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. AD - Center for Innovative Technology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt-Ingram Cancer Center, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, United States. FAU - Cliffel, David E AU - Cliffel DE AD - Department of Chemistry, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute of Chemical Biology, Vanderbilt University, Nashville, TN, United States. AD - Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, Nashville, TN, United States. LA - eng GR - R01 HD102752/HD/NICHD NIH HHS/United States PT - Journal Article DEP - 20210827 PL - Switzerland TA - Front Bioeng Biotechnol JT - Frontiers in bioengineering and biotechnology JID - 101632513 PMC - PMC8431803 OTO - NOTNLM OT - electrochemistry OT - mass spectrometry OT - organ-on-a-chip OT - organophosphate OT - pesticide COIS- The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. EDAT- 2021/09/14 06:00 MHDA- 2021/09/14 06:01 PMCR- 2021/01/01 CRDT- 2021/09/13 06:55 PHST- 2020/10/27 00:00 [received] PHST- 2021/07/16 00:00 [accepted] PHST- 2021/09/13 06:55 [entrez] PHST- 2021/09/14 06:00 [pubmed] PHST- 2021/09/14 06:01 [medline] PHST- 2021/01/01 00:00 [pmc-release] AID - 622175 [pii] AID - 10.3389/fbioe.2021.622175 [doi] PST - epublish SO - Front Bioeng Biotechnol. 2021 Aug 27;9:622175. doi: 10.3389/fbioe.2021.622175. eCollection 2021.