[3H]‐RS‐45041‐190: a selective high‐affinity radioligand for I2 imidazoline receptors

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Abstract
1 RS-45041-190 (4-chloro-2-(imidazolin-2-yl)isoindoline) is an I2 imidazoline receptor ligand with the highest affinity and selectivity so far described; [3H]-RS-45041-190 has a tritium atom attached to the 7-position on the isoindoline ring 2 [3H]-RS-45041-190 binding to rat kidney membranes was saturable (Bmax = 223.1 ± 18.4 fmol mg−1 protein) and of high affinity (Kd = 2.71 ±0.59 nM). Kinetic studies revealed that the binding was rapid and reversible, with [3H]-RS-45041-190 interacting with two sites or two affinity states 3 Competition studies showed that 60–70% of [3H]-RS-45041-190 binding (1 nM) was specifically to imidazoline binding sites of the I2 subtype, characterized by high affinity for idazoxan (pIC50 7.85 ±0.03) and cirazoline (pIC50 8.16 ±0.05). The remaining 30–40% was displaced specifically by the monoamine oxidase A inhibitors, clorgyline and pargyline 4 α1 and α2-adrenoceptor, Ii imidazoline, histamine, 5-hydroxytryptamine or dopamine receptor ligands had low affinity suggesting that [3H]-RS-45041-190 did not label receptors of these classes 5 In autoradiography studies, [3H]-RS-45041-190 labelled discrete regions of rat brain corresponding to the distribution of I2 subtypes, notably the subfornical organ, arcuate nucleus, interpeduncular nucleus, medial habenular nucleus and lateral mammillary nucleus, and additional sites in the locus coeruleus, dorsal raphe and dorsomedial hypothalamic nucleus 6 [3H]-RS-45041-190 therefore labels I2 receptors with high affinity, and an additional site which has high affinity for some monoamine oxidase inhibitors.