MUSCARINIC CHOLINERGIC RECEPTORS IN THE RAT CAUDATE-PUTAMEN AND OLFACTORY TUBERCLE BELONG PREDOMINANTLY TO THE M4 CLASS - INSITU HYBRIDIZATION AND RECEPTOR AUTORADIOGRAPHY EVIDENCE

In order to establish the nature of the muscarinic cholinergic receptors present in the rat caudate-putamen and olfactory tubercle, we have combined in situ hybridization histochemistry with oligonucleotide probes and receptor autoradiography with N-[H-3]methyl scopolamine and several subtype-selective antagonists: hexahydro-sila-difenidol, p-fluoro-hexahydro-sila-difenidol, 4-diphenyl-acetoxy-N-methylpiperidine methbromide, AF-DX 116, pirenzepine and methoctramine. In both brain regions, transcripts for the m4 muscarinic receptor subtype were the most abundant, followed by transcripts for the m1 subtype. m2 and m3 transcripts were much less abundant, whereas m5 mRNA was not detected under the present conditions. The binding profiles obtained in these areas were clearly distinct from those obtained in the CA1 layer of the hippocampus and in the pontine nuclei, regions enriched in M1 and M2 sites, respectively. In contrast, they were in good agreement with the characteristics of atyptical muscarinic receptors present in cell lines such as NG108-15, which contains mRNA for the m4 subtype, and PC12. The profiles displayed by some of the compounds used in the present study for cloned m4 receptors expressed in mammalian cells also agree with our results in rat caudate-putamen and olfactory tubercule. Taken together, these facts support the existence, in rat caudate-putamen and olfactory tubercle, of a major population of muscarinic cholinergic receptors belonging to the M4 type.