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Journal of Neuroscience, Vol 8, 4646-4652, Copyright © 1988 by Society for Neuroscience
Localization of a family of muscarinic receptor mRNAs in rat brain
NJ Buckley, TI Bonner and MR Brann
Laboratory of Cell Biology, National Institute of Mental Health, Bethesda, MD 20892.
A family of 4 rat muscarinic receptors (m1, m2, m3, and m4) have recently
been cloned and sequenced (Bonner et al., 1987). Since pharmacological
probes that are presently available do not appear to distinguish among 3 of
these muscarinic receptors, we constructed oligonucleotide probes
corresponding to the N-terminal sequences of the muscarinic receptors and
used them to specifically localize m1, m2, m3, and m4 mRNA in sections of
rat brain using in situ hybridization histochemistry. Northern analysis
demonstrated a 3.1 kilobase (kb) m 1 mRNA, a 4.5 kb m3 mRNA, and a 3.3 kb
m4 mRNA in cerebral cortex, striatum, hippocampus, and cerebellum. In situ
hybridization histochemistry indicated a prevalence of m1 mRNA in the
pyramidal cell layer of the hippocampus, the granule cell layer of the
dentate gyrus, the olfactory bulb, amygdala, olfactory tubercule, and
piriform cortex. Caudate putamen and cerebral cortex showed moderate levels
of labeling. m2 mRNA was detectable in the medial septum, diagonal band,
olfactory bulb, and pontine nuclei. m3 and m4 mRNA were also prevalent in
the olfactory bulb and pyramidal cell layer of the hippocampus but were
present only in low levels in the dentate gyrus. m3 mRNA was present in
superficial and deep layers of the cerebral cortex, whereas m4 mRNA was
more evenly distributed with a slightly more intense labeling evident in
the midcortical layer. In addition, m3 mRNA was present in a number of
thalamic nuclei and brain-stem nuclei, while m4 mRNA predominated in the
caudate putamen. These data offer a new basis on which to interpret the
heterogeneity of muscarinic actions in the CNS.
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