Dependence of dopamine calibration factors on media Ca2+ and Mg2+ at carbon-fiber microelectrodes used with fast-scan cyclic voltammetry.

Carbon-fiber microelectrodes and voltammetric methods have been used extensively for the detection of dopamine in brain tissue in vivo and in vitro. Voltammetric microelectrodes are often calibrated in non-physiological media, like phosphate-buffered saline, rather than in oxygenated physiological media. Here, we determined dopamine calibration factors (nA microM-1) in several defined solutions for two types of carbon-fiber electrode used with fast-scan cyclic voltammetry. For both electrode types, dopamine calibration factors, and thus electrode sensitivities, were 2-3-fold higher in phosphate- or HEPES-buffered saline than in a bicarbonate-based artificial CSF (ACSF) that reflected that normal ionic composition of brain extracellular fluid. Removal of Ca2+ and Mg2+ from ACSF eliminated this difference. Because extracellular Ca2+ concentration ([Ca2+]o) can fall under stimulation conditions used to elicit dopamine release, we also evaluated the size of stimulated [Ca2+]o shifts in guinea pig midbrain slices using ion-selective microelectrodes. The [Ca2+]o decreases were less than 100 microM, which was well below the mM decreases observed to alter DA sensitivity. Consequently, calibration data obtained in normal physiological solutions should be valid under conditions of mild stimulation. Moreover, calibration in divalent cation-free media will cause calculated DA levels to be underestimated and should be avoided, unless appropriate for a given experimental paradigm.