Preparation and Measuring of Brain GABAAR-coupled Cl-/HCO3-- Activity for Integral Assessment of Aquatic Toxicity

Abstract

The wide use of aromatic hydrocarbons in various industries is having a negative effect on the environment and human health. Therefore, a key focus of current toxicology is the development and use of protein reporters with high sensitivity to various aromatic hydrocarbons (including phenolics and drugs). One molecular target for a wide range of pharmacology drugs and aromatic hydrocarbons (including phenol) is the neuronal GABA_AR-coupled Cl^-/HCO₃^--ATPase. In this study, we present a protocol for isolation of the membrane-bound Cl^-/HCO₃^--ATPase from neuronal cells of animal brain. We then describe an uncomplicated in vitro method for measuring this ATPase activity for assessment of toxicity after interaction of this protein with an aquatic sample. This assay offers new avenues for using the Cl^-/HCO₃^--ATPase as a biomarker of water toxicity. This biotest is efficient, requires very little of the enzyme, and retains its sensitivity at low levels of various compounds. © 2019 by John Wiley & Sons, Inc.