title: Subacute developmental exposure of zebrafish to the organophosphate pesticide metabolite, chlorpyrifos-oxon, results in defects in Rohon-Beard sensory neuron development.
abstract: Organophosphate pesticides (OPs) are environmental toxicants known to inhibit the catalytic activity of acetylcholinesterase (AChE) resulting in hypercholinergic toxicity symptoms. In developing embryos, OPs have been hypothesized to affect both cholinergic and non-cholinergic pathways. In order to understand the neurological pathways affected by OP exposure during embryogenesis, we developed a subacute model of OP developmental exposure in zebrafish by exposing embryos to a dose of the OP metabolite chlorpyrifos-oxon (CPO) that is non-lethal and significantly inhibited AChE enzymatic activity compared to control embryos (43% at 1 day post-fertilization (dpf) and 11% at 2dpf). Phenotypic analysis of CPO-exposed embryos demonstrated that embryonic growth, as analyzed by gross morphology, was normal in 85% of treated embryos. Muscle fiber formation was similar to control embryos as analyzed by birefringence, and nicotinic acetylcholine receptor (nAChR) cluster formation was quantitatively similar to control embryos as analyzed by α-bungarotoxin staining. These results indicate that partial AChE activity during the early days of zebrafish development is sufficient for general development, muscle fiber, and nAChR development. Rohon-Beard (RB) sensory neurons exhibited aberrant peripheral axon extension and gene expression profiling suggests that several genes responsible for RB neurogenesis are down-regulated. Stability of CPO in egg water at 28.5 °C was determined by HPLC-UV-MS analysis which revealed that the CPO concentration used in our studies hydrolyzes in egg water with a half-life of 1 day. The result that developmental CPO exposure affected RB neurogenesis without affecting muscle fiber or nAChR cluster formation demonstrates that zebrafish are a strong model system for characterizing subtle neurological pathologies resulting from environmental toxicants.
authors.affiliation: Department of Biomedical and Pharmaceutical Sciences, University of Montana, Missoula, MT 59812, USA. saskia1.jacobson@umontana.edu
authors.forename: Saskia M
authors.initials: SM
authors.lastname: Jacobson
authors.forename: Denise A
authors.initials: DA
authors.lastname: Birkholz
authors.forename: Marcy L
authors.initials: ML
authors.lastname: McNamara
authors.forename: Sandip B
authors.initials: SB
authors.lastname: Bharate
authors.forename: Kathleen M
authors.initials: KM
authors.lastname: George
mesh_terms: D000818:Animals; D004390:Chlorpyrifos / Q000031:analogs & derivatives* / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004625:Embryo, Nonmammalian / Q000002:abnormalities / Q000187:drug effects / Q000648:ultrastructure; D047108:Embryonic Development / Q000187:drug effects; D015870:Gene Expression / Q000187:drug effects; D007306:Insecticides / Q000633:toxicity*; D055495:Neurogenesis / Q000187:drug effects / Q000235:genetics; D009469:Neuromuscular Junction / Q000187:drug effects / Q000254:growth & development; D010755:Organophosphates / Q000633:toxicity; D011984:Sensory Receptor Cells / Q000187:drug effects* / Q000648:ultrastructure; D018675:Toxicity Tests; D014874:Water Pollutants, Chemical / Q000633:toxicity; D015027:Zebrafish / Q000002:abnormalities / Q000196:embryology / Q000254:growth & development*
chemical_list: D007306:Insecticides; D010755:Organophosphates; D014874:Water Pollutants, Chemical; C009618:O,O-diethyl O-3,5,6-trichloro-2-pyridyl phosphate; D004390:Chlorpyrifos