Detecting zinc release induced by mu-opioid receptor agonists in brain slices

Abstract

After iron, zinc is the second most abundant transition metal in living organisms and it is known to be a contributory factor in a series of neurological disorders. In biological systems zinc exists as either bound Zn2+, representing the majority of the total zinc in tissues, or free (chelatable) Zn2+. Several fluorescents dyes have been developed to detect free zinc when it is released from zinc-binding proteins, which occurs via redox mechanisms in response to the stimulation of a number of neurotransmitter receptors, including the μ opioid receptor. Here we describe a detailed protocol to detect drug stimulated intracellular zinc release in rodent brain slices using time-lapse microscopy and fluorescence imaging.