Receptor Localization Using [3Hmesulergine

In 1983, having just set up the autoradiographic methodology for receptor localization developed in Mike Kuhar's laboratory at the Johns Hopkins University to label brain tissue sections with radioligands, Jose M. Palacios decided to investigate the localization of [3H]mesulergine. At his lab, Rolf Lenherr incubated rat brain tissue sections with the radiolabeled ligand and generated some autoradiograms. When we developed the films, we were disappointed - at first glance the film appeared "empty." A closer look revealed some repeated dark lines in the film; after a while, we realized that those heavy labeled thready-looking spots corresponded to the choroid plexus (Fig. 1.1) and were filling the brain ventricles in our tissues sections. This labeling was selectively blocked by coincubation with unlabeled serotonin. We knew from our own experience that radiolabeled serotonin also labeled intensely the choroid plexuses in the rat brain, as well as many other brain areas, but no special attention had been paid to them.

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Fig. 1.1 Images from early autoradiograms of sagittal and horizontal rat brain sections incubated with [3H]mesulergine [(a), (b), and (c)] or 3H-5-HT [(d), (e), and (f)] and different unlabeled compounds. (a) Short exposure revealing intense labeling in choroid plexus. (b) and (c) are longer exposures showing binding of [3H]mesulergine to 5-HT2A and 5-HT2C; (b) and (c) show the blockade of 5-HT2A binding by coincubation with ketanserin. (d) shows [3H]5-HT binding to 5-HT1A, 5-HT1B and 5-HT2C. (e) Binding to 5-HT1A has been blocked with 8-OH-DPAT. (f) 5-HT1B binding was blocked with compound SDZ 21009 (see text for details)

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