Collecting Pig Choroid Plexus

Since the putative 5-HT receptor had been clearly identified in the choroid plexus (we later demonstrated its presence in other brain regions but to a much lesser extent), we decided to study its pharmacology by applying radioligand binding to membranes of choroid plexus. As the rat choroid plexus are not easy to collect in large quantities in a reasonable manner, we turned to pig brains - it was cheaper to collect the choroid plexus from brains obtained from the local slaughterhouse. However, dealing with tissue preparation was not simple, since the choroid plexus is primarily a tight mix of vessels and harsh connective tissue, but with the initial help of Monique Rigo, a resourceful technician at JMP laboratory, we managed to obtain good enough membrane fractions, perform regular membrane labeling studies, and get decent quality binding data (Pazos et al. 1984a, b; Hoyer et al. 1985a).

From there, we started parallel autoradiographic studies to establish what came to be the 5-HT2C receptor, as will be discussed later in section 1.4 in this chapter. The project in fact followed a research plan carried out in parallel in two different buildings: Autoradiographic experiments were done at building 360 (JMP lab), while membrane assays were carried out at building 386 (DH lab). During the autumn of 1983 and the whole year 1984, it become quite usual to Angel to walk daily from one place to the other, bearing tissue samples, autoradiograms and counter prints in his hands, and frequently, a certain level of anxiety in his mind, since we were using old-generation liquid scintillation counters, with rather modest throughput, when we were preparing hundreds of samples. This meant that they had to be loaded and unloaded one by one, by hand with two or three loading sessions per day, weekends included; every sample was labeled by hand in case of counter failure and to allow recounting.

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