Please use this identifier to cite or link to this item: http://ahro.austin.org.au/austinjspui/handle/1/13633
Title: Pharmacology and subcellular distribution of [3H]rilmenidine binding sites in rat brain.
Authors: Hosseini, A R;Jackman, G P;King, P R;Louis, William J;Gundlach, Andrew L
Affiliation: The University of Melbourne, Department of Medicine, Austin and Repatriation Medical Centre, Heidelberg, Victoria, Australia.
Issue Date: 15-Oct-1998
Citation: Journal of the Autonomic Nervous System; 72(2-3): 129-36
Abstract: We have previously reported that in rat brain membranes, [3H]rilmenidine, in addition to labelling alpha2-adrenoceptors and the I2B-subtype of imidazoline receptor binding site (I2B-RBS), may label an additional I-RBS population, distinct from previously classified I1-RBS and I2-RBS. In this study, using crude or fractionated rat brain membranes we examined the possible association of [3H]rilmenidine-labelled I-RBS with the A- and B-isoforms of monoamine oxidase (MAO) by studying the inhibition of [3H]rilmenidine binding by a number of MAO inhibitors; and comparing the maximal binding density (Bmax) and subcellular distribution of [3H]rilmenidine binding sites with that of MAO-A and MAO-B catalytic sites labelled by [3H]RO41-1049 and [3H]RO19-6327 and 12-RBS labelled by [3H]2-BFI. Inhibition of [3H]rilmenidine binding by all MAO inhibitors tested produced very shallow curves (slope 0.29-0.56). Clorgyline and moclobemide (selective MAO-A inhibitors) displayed moderate affinities (60-140 nM), while pargyline (non-selective MAO-inhibitor), RO41-1049 (selective MAO-A inhibitor) and RO19-6327 (selective MAO-B inhibitor) exhibited very low affinities (> 2 microM) for 50-75% of [3H]rilmenidine-labelled I-RBS in crude brain membranes and even lower affinity for the remaining binding. Under identical buffer conditions, the Bmax of [3H]rilmenidine-labelled I-RBS (1.45+/-0.14 pmol/mg protein) was considerably lower than those of MAO-A (13.10+/-0.15 pmol/mg) and MAO-B (10.35+/-0.50 pmol/mg) sites. These results suggest that [3H]rilmenidine does not interact directly with the active catalytic site of either MAO enzyme and could at best only associate with a subpopulation of MAO molecules. Binding studies on five fractions of rat cortex homogenates-nuclear (N), heavy (M) and light (L) mitochondrial, microsomal non-mitochondrial (P), and soluble cytosolic (S) fractions-revealed that 45% of total [3H]rilmenidine binding was present in the P fraction cf. 20 and 23% in the M and L fractions, in contrast to [3H]RO19-6327 and [3H]2-BFI which bound 11-13% in the P fraction and 36-38% and 35-44% in the M and L fractions, respectively. Binding of all ligands in the N fraction was 6-15% of total. These studies reveal that [3H]rilmenidine-labelled I-RBS, unlike the I2-RBS, are not predominantly associated with mitochondrial fractions containing the MAO enzymes (and cytochrome oxidase activity), but appear to be distributed in both the mitochondrial and plasma membrane fractions in rat cerebral cortex.
Internal ID Number: 9851561
URI: http://ahro.austin.org.au/austinjspui/handle/1/13633
URL: http://www.ncbi.nlm.nih.gov/pubmed/9851561
Type: Journal Article
Subjects: Adrenergic alpha-Agonists.metabolism
Animals
Binding Sites
Brain.drug effects.enzymology.metabolism.ultrastructure
Imidazoles.metabolism
Imidazoline Receptors
In Vitro Techniques
Male
Membranes
Monoamine Oxidase.metabolism
Monoamine Oxidase Inhibitors.pharmacology
Oxazoles.metabolism
Picolinic Acids.metabolism
Radioligand Assay
Rats
Rats, Inbred WKY
Receptors, Drug.antagonists & inhibitors.metabolism
Subcellular Fractions.drug effects.enzymology.metabolism
Thiazoles.metabolism
Appears in Collections:Journal articles

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