Characterization of Tritiated Noradrenaline Release from the Rat Preoptic Area with Microdialysis In Vivo

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Abstract
Present techniques are unable to provide a sensitive and accurate index of noradrenergic activity in the rat preoptic area. In this study, we have examined the brainstem A1 noradrenergic input to the preoptic area using a new technique whereby [3H]noradrenaline is preloaded into the preoptic area and release of radioactivity from this region is measured subsequently using microdialysis in vivo. Electrical stimulation of the ipsilateral A1 area for 20 min at 5, 10, and 15 Hz evoked significant increases in dialysate radioactivity that were repeatable and frequency-dependent. After removal of calcium from the perfusion medium, basal release of radioactivity was markedly reduced and the effect of A1 stimulation abolished. Changing to a 100 mM K+ medium evoked an increase in the release of radioactivity that was sixfold greater than that seen after A1 stimulation. Separation of the dialysate with HPLC showed that 33% of the increase in measured radioactivity after A1 stimulation was directly attributable to [3H]noradrenaline and the remainder to the metabolites vanillylmandelic acid, 3,4-dihydroxymandelic acid, and 3,4-dihydroxyphenylglycol. In contrast, the increase in radioactivity after K+ depolarization was due almost completely to [3H]noradrenaline. Addition of 10 microM clonidine to the perfusion medium markedly reduced basal release of radioactivity, but had no effect on evoked release following A1 stimulation. Conversely, perfusion with 10 microM yohimbine had no effect on basal release, but significantly increased evoked release after A1 stimulation. These results now provide a characterization of noradrenergic activity in the preoptic area and indicate the importance of the A1 noradrenergic input to this region. The technique of measuring radioactivity with microdialysis after preloading with [3H]noradrenaline provides a relatively simple, sensitive index of noradrenergic activity in vivo with good temporal resolution.