Characterization of probe and tissue factors that influence interpretation of quantitative microdialysis experiments for dopamine.
Two quantitative methods, the Lönnroth (no-net-flux) and variation of perfusion flow rate methods, were used to investigate the influence of the probe and tissue on dopamine microdialysis measurements. In vivo measurements were made in the nucleus accumbens of awake, freely moving rats on two consecutive days of dialysis. The results of the no-net-flux study showed that there was no statistically significant difference in extraction fraction at a perfusion flow rate of 2.0 microl/min between in vitro in a well-stirred solution and in vivo measured during 2 days of continuous dialysis. Also, varying the perfusate flow rate over the range 0.25-2.0 microl/min produced a variation in the extraction fraction that was the same in vitro and in vivo. These results indicate that the extraction fraction for dopamine over the 2 days was dominated by the properties of the probe. The negligible influence of the tissue on dopamine extraction fraction was probably due to the high basal activity of the dopamine transporter in vivo. Therefore, the extraction fraction is unlikely to be sensitive to increases in dopamine uptake in the vicinity of the probe. The apparent extracellular dopamine concentration increased by 37% on the second day of dialysis while the calcium-dependence of basal dialysate dopamine levels declined by 20%. These findings are consistent with a decrease in physiological viability of the dopamine nerve terminals surrounding the probe during a long-term experiment.
Tang
Amanda
A
Division of Pharmacology and Toxicology, College of Pharmacy, University of Texas at Austin, Austin, TX 78712, USA. matang@mail.utexas.edu
Bungay
Peter M
PM
Gonzales
Rueben A
RA
D000818:Animals; D002118:Calcium / Q000737:chemistry; D004298:Dopamine / Q000032:analysis*; D019544:Equipment Failure Analysis / Q000379:methods*; D005110:Extracellular Space / Q000737:chemistry / Q000378:metabolism*; D008297:Male; D017551:Microdialysis / Q000379:methods*; D015335:Molecular Probes; D009714:Nucleus Accumbens / Q000737:chemistry / Q000166:cytology / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D015335:Molecular Probes; D002118:Calcium; D004298:Dopamine