Journal ArticleDOI
m1-m5 muscarinic receptor distribution in rat CNS by RT-PCR and HPLC.
TLDR
The combination of RT‐PCR and HPLC provides a rapid and sensitive method for quantifying the expression of mRNA coding for all five muscarinic receptor subtypes derived from the CNS.Abstract:
Five muscarinic receptor genes (m1-m5) that encode distinct muscarinic receptor subtypes have been cloned. Because of their structural homology and pharmacological similarity, ligand binding probes currently available do not clearly distinguish among the subtypes. To obtain a clear distribution within the CNS of molecularly defined muscarinic receptor subtypes, seven brain regions were examined for the expression of the respective mRNAs. The most sensitive method for detecting mRNA is through amplification of the respective cDNAs. Brain regions were obtained from male Wistar rats, and total RNA was isolated. The isolates were extensively treated with RNase-free DNase to remove any residual genomic DNA. Total RNA (1 microgram) was reverse-transcribed using random primers and reverse transcriptase. The resulting cDNA was amplified using a thermal cycler, and the polymerase chain reaction (PCR)-amplified products were analyzed by gel electrophoresis containing ethidium bromide and visualized with fluorescent illumination. PCR-amplified samples were also injected directly onto an HPLC anion exchange column and quantified by UV detection. Each of the five muscarinic subtypes was found in every brain region examined. The m1 subtype was most abundant in cortex and gradually declined in content caudally to the spinal cord. The m2 subtype was most abundant in thalamus-hypothalamus and ponsmedulla. The m4 subtype was found in greatest amount in the striatum, whereas m3 and m5 were expressed consistently throughout the CNS. The combination of RT-PCR and HPLC provides a rapid and sensitive method for quantifying the expression of mRNA coding for all five muscarinic receptor subtypes derived from the CNS.read more
Citations
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Journal ArticleDOI
Selective cognitive dysfunction in acetylcholine M1 muscarinic receptor mutant mice.
Stephan G. Anagnostaras,Geoffrey G. Murphy,Susan E. Hamilton,Scott L. Mitchell,Nancy P. Rahnama,Neil M. Nathanson,Alcino J. Silva +6 more
TL;DR: Memory in mice with a null mutation of the gene coding the M1 receptor, the most densely distributed muscarinic receptor in the hippocampus and forebrain is examined, suggesting that the M 1 receptor is specifically involved in memory processes for which the cortex and hippocampus interact.
Journal ArticleDOI
M1 Receptors Play a Central Role in Modulating AD-like Pathology in Transgenic Mice
Antonella Caccamo,Salvatore Oddo,Lauren M. Billings,Kim N. Green,Hilda Martinez-Coria,Abraham Fisher,Frank M. LaFerla +6 more
TL;DR: AF267B represents a peripherally administered low molecular weight compound to attenuate the major hallmarks of AD and to reverse deficits in cognition and selective M1 agonists may be efficacious for the treatment of AD.
Journal ArticleDOI
Pteropodine and isopteropodine positively modulate the function of rat muscarinic M1 and 5-HT2 receptors expressed in Xenopus oocyte
Tai-Hyun Kang,Kinzo Matsumoto,Michihisa Tohda,Yukihisa Murakami,Hiromitsu Takayama,Mariko Kitajima,Norio Aimi,Hiroshi Watanabe +7 more
TL;DR: It is suggested that pteropodine and isopteropodines act as positive modulators of muscarinic M(1) and 5-HT(2) receptors.
Journal ArticleDOI
Nicotinic receptor-mediated effects on appetite and food intake.
TL;DR: An overview of the literature at, or near, the interface of nicotinic receptors and appetite regulation is presented and the hypothesis that the lateral hypothalamus is a particularly important locus of the anorectic effects of nicotine is presented.
Journal ArticleDOI
Cholinergic dilation of cerebral blood vessels is abolished in M 5 muscarinic acetylcholine receptor knockout mice
Masahisa Yamada,Kathryn G. Lamping,Alokesh Duttaroy,Weilie Zhang,Yinghong Cui,Frank P. Bymaster,David L. McKinzie,Christian C. Felder,Chu-Xia Deng,Frank M. Faraci,Jürgen Wess +10 more
TL;DR: In vitro neurotransmitter release experiments showed that M5 receptors play a role in facilitating muscarinic agonist-induced dopamine release in the striatum, providing direct evidence that M1-M5 receptors are physiologically relevant.
References
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Journal ArticleDOI
Identification of a family of muscarinic acetylcholine receptor genes
TL;DR: Analysis of human and rat genomic clones indicates that there are at least four functional muscarinic receptor genes and that these genes lack introns in the coding sequence.
Journal ArticleDOI
Identification and localization of muscarinic acetylcholine receptor proteins in brain with subtype-specific antibodies
TL;DR: The results clarify the roles of these genetically defined receptor proteins in cholinergic transmission in brain and suggest that 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.
Journal ArticleDOI
Cloning, sequencing and expression of complementary DNA encoding the muscarinic acetylcholine receptor
Tai Kubo,Kazuhiko Fukuda,Atsushi Mikami,Akito Maeda,Hideo Takahashi,Masayoshi Mishina,Tatsuya Haga,Kazuko Haga,Arata Ichiyama,Kenji Kangawa,Masayasu Kojima,Hisayuki Matsuo,Tadaaki Hirose,Shosaku Numa +13 more
TL;DR: Cloning and sequence analysis of DNA complementary to porcine cerebral messenger RNA encoding the muscarinic acetylcholine receptor predict the complete amino-acid sequence of this protein.
Journal ArticleDOI
Distinct primary structures, ligand-binding properties and tissue-specific expression of four human muscarinic acetylcholine receptors.
TL;DR: Differences among subtypes in the affinities and proportions of such sites suggest the capacity of mAChR subtypes to interact differentially with the cellular effector‐coupling apparatus.
Journal ArticleDOI
Cloning and expression of the human and rat m5 muscarinic acetylcholine receptor genes.
TL;DR: expression of mRNA has yet to be observed in brain or selected peripheral tissues, suggesting that either it is substantially less abundant than m1-m4 or its distribution is quite different.