Construction of 1 mm microdialysis probe for amino acids dialysis in rats

We describe here a microdialysis probe with 1 mm opening for precise and confined dialysis area in the awake, freely moving rat. This probe is designed to allow the local diffusion of the perfusion medium to an area approximately 175 μm high, 266 μm wide (mediolateral direction), and 305 μm in rostrocaudal direction. In addition, the probe allows the local application of drugs to the same precise area of interest. The probe was constructed from a piece of 25 gauge tubing with 1 mm hallowed opening located 0.5 mm from the distal (inserting) end. The dialysis fiber which was inserted into the stainless steel 25 gauge tubing and cemented into place has 200 μm diameter and 5000 molecular weight cut off. We tested the probe diffusion extent by direct infusion of fluorogold through the dialysis cannula. Changes in the extracellular concentrations of amino acids were measured in response to infusion of veratridine a sodium channel activator. All amino acids tested showed a significant 80% times decrease in their recovery concentration when compared to their respective concentrations recovered through 2 mm probe constructed earlier in our laboratory (Renno et al., 1992). Tests in awake rats with probes in the ventrocaudal PAG showed stable amounts of 12 different amino acids during repeated (6-8 times) 12 min samples at 3-5 μl/min collecting rate. Depolarization with 75 μM veratridine resulted in significant elevation in extracellular γ- aminobutyric acid (GABA), aspartate, glutamate, taurine, glycine and citrulline. This design enables us to apply drugs of interest and measure the concentrations of amino acid neurotransmitters to a more precise, delineated and premeasured areas in the CNS.