Repeated perfusion with elevated potassium in in vivo microdialysis--A method for detecting small changes in extracellular dopamine.

As a great deal of variability between subjects is often seen when using the microdialysis technique to measure the effects of depolarizing agents on extracellular neurotransmitter levels, we have developed a technique to account for the variability inherent in this method. High potassium (50 or 100 mM) artificial cerebrospinal fluid (ACSF), perfused through the probe for 10 min, significantly increased extracellular dopamine (DA) concentration during both an initial and second perfusion, and the two responses were highly correlated. However, extracellular DA returned to normal following the first perfusion with 50 mM K+ but not 100 mM K+ perfusion. The slope of the regression line obtained by plotting the response of the second K+ perfusion as a function of the first K+ perfusion for all K+ concentrations was 1.03 (not significantly different from unity). Similarly, when the time between two 50 mM potassium perfusions was varied from 30-150 min, the responses were highly correlated. This technique was used to demonstrate an interaction between N-methyl-D-aspartate (NMDA) and 50 mM K+. Perfusion of 0.1 mM NMDA alone had no effect on extracellular DA, but NMDA paired with a 50 mM K+ perfusion significantly increased extracellular DA over that increase by 50 mM K+ alone. We propose that a first stimulation with 50 mM potassium may characterize an individual animal's responsiveness to a depolarizing stimulus, and may be used as a control for testing drug effects by coupling drug treatments with a second 50 mM potassium stimulation to give a more accurate measure of small changes in extracellular dopamine.