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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-[³H]methyl scopolamine and several subtype-selective antagonists: hexahydro-sila-difenidol, p-fluoro-hexahydro-sila-difenidol, 4-diphenylacetoxy-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 …