TY - JOUR
T1 - Identification and localization of muscarinic acetylcholine receptor proteins in brain with subtype-specific antibodies
AU - Levey, A. I.
AU - Kitt, C. A.
AU - Simonds, W. F.
AU - Price, D. L.
AU - Brann, M. R.
PY - 1991
Y1 - 1991
N2 - mRNAs encoding five genetically distinct muscarinic ACh receptors are present in the CNS. Because of their pharmacological similarities, it has not been possible to detect the individual encoded proteins; thus, their physiological functions are not well defined. To characterize the family of proteins, a panel of subtype-selective antibodies was generated against recombinant muscarinic receptor proteins and shown to bind specifically to each of the cloned receptors. Using immunoprecipitation, three receptor proteins (m1, m2, and m4) accounted for the vast majority of the total solubilized muscarinic binding sites in rat brain. These receptor subtypes had marked differences in regional and cellular localization as shown by immunocytochemistry. The m1-protein was present in cortex and striatum and was localized to cell bodies and neurites, consistent with its role as a major postsynaptic muscarinic receptor. The m2-receptor protein was abundant in basal forebrain, scattered striatal neurons, mesopontine tegmentum, and cranial motor nuclei; this distribution is similar to that of cholinergic neurons and suggests that m2 is an autoreceptor. However, m2 was also present in noncholinergic cortical and subcortical structures, providing evidence that this subtype may presynaptically modulate release of other neurotransmitters and/or function postsynaptically. The m4-receptor was enriched in neostriatum, olfactory tubercle, and islands of Calleja, indicating an important role in extrapyramidal function. These results clarify the roles of these genetically defined receptor proteins in cholinergic transmission in brain. Since the nonselective muscarinic drugs used in the treatment of patients with neurological disease produce many side effects, the characterization of receptor subtypes, including cellular and subcellular localization, will be of great value in defining the targets for the development of more effective and specific therapeutic agents.
AB - mRNAs encoding five genetically distinct muscarinic ACh receptors are present in the CNS. Because of their pharmacological similarities, it has not been possible to detect the individual encoded proteins; thus, their physiological functions are not well defined. To characterize the family of proteins, a panel of subtype-selective antibodies was generated against recombinant muscarinic receptor proteins and shown to bind specifically to each of the cloned receptors. Using immunoprecipitation, three receptor proteins (m1, m2, and m4) accounted for the vast majority of the total solubilized muscarinic binding sites in rat brain. These receptor subtypes had marked differences in regional and cellular localization as shown by immunocytochemistry. The m1-protein was present in cortex and striatum and was localized to cell bodies and neurites, consistent with its role as a major postsynaptic muscarinic receptor. The m2-receptor protein was abundant in basal forebrain, scattered striatal neurons, mesopontine tegmentum, and cranial motor nuclei; this distribution is similar to that of cholinergic neurons and suggests that m2 is an autoreceptor. However, m2 was also present in noncholinergic cortical and subcortical structures, providing evidence that this subtype may presynaptically modulate release of other neurotransmitters and/or function postsynaptically. The m4-receptor was enriched in neostriatum, olfactory tubercle, and islands of Calleja, indicating an important role in extrapyramidal function. These results clarify the roles of these genetically defined receptor proteins in cholinergic transmission in brain. Since the nonselective muscarinic drugs used in the treatment of patients with neurological disease produce many side effects, the characterization of receptor subtypes, including cellular and subcellular localization, will be of great value in defining the targets for the development of more effective and specific therapeutic agents.
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U2 - 10.1523/jneurosci.11-10-03218.1991
DO - 10.1523/jneurosci.11-10-03218.1991
M3 - Article
C2 - 1941081
AN - SCOPUS:0025943814
SN - 0270-6474
VL - 11
SP - 3218
EP - 3226
JO - Journal of Neuroscience
JF - Journal of Neuroscience
IS - 10
ER -