Table 1.
Experimental parameters for receptor/ transporters binding assays
| Receptor/transporter | Brain tissue | Radioligand (concentration) | Nonspecific displacer | Ionic conditions | Incubation parameters |
|---|---|---|---|---|---|
| 5-HT1A | cerebral cortex | [3H]-WAY-100,635 (3.0 nm) | 8-OHDPAT | 60 min, ambient T°, in the presence of 10 nm L-745,870 | |
| 5-HT2A | cerebral cortex | [3H]-ketanserin (2.5 nm) | spiperone | 45 min, 4°C | |
| NET | cerebellum | [3H]-nisoxetine (2.0 nm) | maprotiline | 300 mm NaCl, 5 mm KCl | 60 min, 4°C |
| SERT | ROB | [3H]-citalopram (2.5 nm) | paroxetine | 120 mm NaCl, 5 mm KCl | 60 min, ambient T° |
| DAT | striatum | [3H]-GBR 12935 (5.6 nm) | GBR 12909 | 125 mm NaCl | 45 min, ambient T° |
Brain tissue concentration remained fixed for all of the experiments (2 mg/ml of protein). The nonspecific displacer concentration remained fixed for all of the experiments (10 μm). For 5-HT1A receptor binding, 10 nm L-745,870 was added to each well to prevent [3H]-WAY-100,635 from interacting with dopaminergic D4 receptors (Chemel et al., 2006). Such a concentration of L-745,870 was chosen because it is >20-fold over the Ki for D4 receptors (0.43 nm), while being >10 times under the Ki of L-745,870's next target (sigma receptors, 130 nm) (Patel et al., 1997). The remainder of brain (ROB) corresponds to the brain areas remaining following dissection of the cortex, striatum, and cerebellum. This encompasses the raphe nuclei, substantia nigra, diencephalon, globus pallidus, hippocampus, and subcortical white matter. Ambient T°, 20–22°C.