Quantitative autoradiographic localization in rat brain of alpha 2-adrenergic and non-adrenergic I-receptor binding sites labelled by [3H]rilmenidine

King, P.R.; Gundlach, A.L.; Louis, W.J.

Brain Research 675(1-2): 264-278

1995


ISSN/ISBN: 0006-8993
PMID: 7796138
DOI: 10.1016/0006-8993(95)00083-3
Accession: 044113817

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Abstract
alpha 2A-Adrenergic receptor (AR) and non-adrenergic imidazoline receptor (I-R) binding sites have been previously characterized in rat cerebral cortex membranes using the N-substituted oxazoline, [3H]rilmenidine ([3H]Ril) [King, P.R. et al., Eur. J. Pharmacol., 218 (1992) 101-108]. In the present study, in vitro autoradiography was used to quantify the regional distribution of these receptors throughout the rat neuroaxis. The distribution and relative density (fmol/mg tissue) of I-Rs was examined in the presence of 1 microM adrenaline to block the adrenergic component of 40 nM [3H]Ril binding and non-specific binding was measured in the presence of another oxazoline, Bay a6781 (10 microM). Both alpha 2A-ARs and I-Rs were broadly, but heterogeneously, distributed. In forebrain, high levels of [3H]Ril-labelled alpha 2A-AR sites were observed in the anterior olfactory nucleus, the piriform, entorhinal and perirhinal cortices, lateral septum, bed nucleus of the stria terminalis, several thalamic nuclei, the amygdala and the arcuate, dorsomedial and posterior hypothalamic nuclei. In hindbrain, alpha 2A-AR sites were concentrated in locus coeruleus, lateral parabrachial nucleus, nucleus of the solitary tract and area postrema. I-R sites accounted for 50% or more of specific [3H]Ril binding (40 nM) in most cortical and hypothalamic nuclei, nucleus of the solitary tract, cranial motor nuclei and most spinal cord layers. The highest densities of I-Rs were found in the arcuate, dorsomedial and posterior hypothalamic nuclei, the locus coeruleus, the area postrema, the cranial motor nuclei and associated with spinal motor neurones. A very high concentration of I-Rs was also detected in the pineal gland. The distribution of alpha 2-AR sites determined resembled that reported with [3H]p-aminoclonidine which appears to specifically label alpha 2-ARs and not I1-R sites in rat brain sections, and [3H]methoxyidazoxan which is a selective alpha 2-AR antagonist. The regional and cellular distribution of I-R binding sites was unlike the distribution of putative I1-R sites labelled by [3H]clonidine in human brain, although comparable autoradiographic mapping studies in rat brain have not been done using this ligand. The regional and cellular distribution of [3H]-labelled I-R binding sites had both similarities and differences to that reported using the imidazoline ligand, [3H]idazoxan, with common labelling of areas such as area postrema, arcuate and interpeduncular nuclei and pineal gland with the two ligands, and differential relative binding levels ([3H]Ril > [3H]idazoxan) associated with hippocampal pyramidal cells and brainstem and spinal motor neurones.