Radioligand binding assay of M1–M5 muscarinic cholinergic receptor subtypes in human peripheral blood lymphocytes

Tayebati, Seyed Khosrow; Codini, Michela; Gallai, Virgilio; Mannino, F; Parnetti, Lucilla; Rícci, Alberto; Sarchielli, Paola; Amenta, Francesco

doi:10.1016/s0165-5728(99)00119-8

Cited by 37 publications

(19 citation statements)

References 28 publications

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“…49 Additionally, the recent discovery of the h4 receptor primarily expressed in bone marrow and eosinophils, but undetected in brain tempts speculation of a peripheral immune target particularly with respect to asthma and allergy, to which clozapine has moderate affinity. 50 The current data confirm and extend on previous pharmacological 26,27 and mRNA expression studies that suggested the existence of multiple muscarinic receptors on blood lymphocytes. 33,63,51,52 While mRNA for m3, m4, and m5 receptors has been reported in mononuclear cells, the additional detection of m3 and m4 mRNA in neutrophils and stromal cells remains a novel finding.…”

Section: Discussionsupporting

confidence: 89%

“…20 In addition, peripheral cells such as blood lymphocytes express D2, 21 D3, 22,23 D4, 23,24 D5 receptors, 25 M2, M3, M4, M5 receptors 26,27 and a1 28 and a2 receptors 29,30 using radioligand binding studies. Since pharmacological characterization remains difficult given the lack of suitable receptor-specific ligands, 31 the expression of neurotransmitter receptors by the detection of corresponding mRNAs has recently been undertaken.…”

Section: Introductionmentioning

confidence: 99%

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Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

et al. 2003

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The antipsychotic drug clozapine, acts via interaction with selective neurotransmitter receptor systems. Its use however, is associated with lifethreatening agranulocytosis. The mechanism by which this occurs and its possible relationship with the drug's atypicality remain unclear. As a first step in identifying mechanistic pathways involved, profiling of neurotransmitter receptors on human neutrophils, mononuclear and bone marrow stromal cells as putative targets for clozapine-mediated toxicity was undertaken. Expression of mRNA encoding dopaminergic d2, d3, d4; serotonergic 5ht2a, 5ht2c, 5ht3, 5ht6, 5ht7; adrenergic a1a, a2; histaminergic h1 and muscarinic m1, m2, m3, m4, m5 receptors was analyzed by reverse transcription-polymerase chain reaction methods. While 5ht2c, 5ht6, m1 and m2 mRNA were undetected, the presence of the other receptors indicates sites at which clozapine could bind and induce toxicity of neutrophils and stromal components which regulate granulopoiesis. The functional significance of differential receptor expression while unknown, may argue for neural regulation of hematopoiesis.

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Section: Discussionsupporting

confidence: 89%

Section: Introductionmentioning

confidence: 99%

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

et al. 2003

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“…Radioligand binding and RT-PCR have identified the presence of mAChRs M2-M5 on PBMCs [19,20] and using RT-PCR, it has been demonstrated that human PBMC express between 3 and 5 of the known muscarinic receptor types including M1 receptors [14]. Interestingly, this last study demonstrated differences in the array of mAChRs (and nAChRs) expressed by PBMCs from different individuals, thus revealing a basis for individual variation of cholinergic immunoregulation.…”

Section: Discussionmentioning

confidence: 74%

“…T cells have also been shown to express choline acetyltransferase mRNA [13], and to synthesise and release ACh [14][15][16]. Furthermore, both T and B lymphocytes isolated from peripheral blood, and Tand B-cell lines, have been reported to express a range of both nicotinic and muscarinic AChRs [17][18][19][20][21]. A limited number of studies suggest that activation of these receptors can signal the augmentation or dimunition of the cell activation process [12,22].…”

Section: Introductionmentioning

confidence: 99%

An in vitro investigation of the effects of the nerve agent pretreatment pyridostigmine bromide on human peripheral blood T-cell function

Telford¹,

Wilkinson²,

Hooi³

et al. 2004

International Immunopharmacology

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“…Interestingly, the use of highly sensitive reverse transcriptase-PCR (RT-PCR) techniques suggests that M 5 receptors are expressed in all major brain regions (12). More recently, M 5 receptors also have been detected in several peripheral tissues or cells including peripheral blood lymphocytes (13,14), skin fibroblasts (15), smooth muscle of the iris sphincter (16) and the esophagus (17), and parotid gland tissue (18), where they are colocalized with several other muscarinic receptor subtypes (13)(14)(15)(16)(17)(18).…”

mentioning

confidence: 99%

Cholinergic dilation of cerebral blood vessels is abolished in M ₅ muscarinic acetylcholine receptor knockout mice

Yamada

Lamping

Duttaroy

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

240

208

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The M5 muscarinic receptor is the most recent member of the muscarinic acetylcholine receptor family (M1-M5) to be cloned. At present, the physiological relevance of this receptor subtype remains unknown, primarily because of its low expression levels and the lack of M 5 receptor-selective ligands. To circumvent these difficulties, we used gene targeting technology to generate M 5 receptor-deficient mice (M5R ؊/؊ mice). M5R ؊/؊ mice did not differ from their wild-type littermates in various behavioral and pharmacologic tests. However, in vitro neurotransmitter release experiments showed that M 5 receptors play a role in facilitating muscarinic agonist-induced dopamine release in the striatum. Because M 5 receptor mRNA has been detected in several blood vessels, we also investigated whether the lack of M 5 receptors led to changes in vascular tone by using several in vivo and in vitro vascular preparations. Strikingly, acetylcholine, a powerful dilator of most vascular beds, virtually lost the ability to dilate cerebral arteries and arterioles in M5R ؊/؊ mice. This effect was specific for cerebral blood vessels, because acetylcholine-mediated dilation of extracerebral arteries remained fully intact in M5R ؊/؊ mice. Our findings provide direct evidence that M 5 muscarinic receptors are physiologically relevant. Because it has been suggested that impaired cholinergic dilation of cerebral blood vessels may play a role in the pathophysiology of Alzheimer's disease and focal cerebral ischemia, cerebrovascular M 5 receptors may represent an attractive therapeutic target. M olecular cloning studies have revealed the existence of five molecularly distinct muscarinic acetylcholine receptor subtypes (M 1 -M 5 ) (1, 2). During the past decade, considerable progress has been made in delineating the physiological roles of the M 1 -M 4 muscarinic receptors (3, 4). In contrast, the physiological relevance of the M 5 receptor subtype, which is the most recent member of the muscarinic receptor family to be cloned (5, 6), remains unknown at present (7,8). However, expression of the cloned M 5 muscarinic receptor gene in cultured mammalian cells has shown that the encoded receptor protein is functional and efficiently couples to G proteins of the G q family, similar to the M 1 and M 3 receptor subtypes (5-8).Immunoprecipitation and in situ mRNA hybridization studies have demonstrated the presence of M 5 receptor protein͞mRNA in different areas of the brain including hippocampus, hypothalamus, and distinct midbrain regions (substantia nigra pars compacta and ventral tegmental area) (9-11). However, M 5 receptors are expressed at very low levels, representing less than 2% of the total muscarinic receptor population (M 1 -M 5 ) expressed in the brain (9). Interestingly, the use of highly sensitive reverse transcriptase-PCR (RT-PCR) techniques suggests that M 5 receptors are expressed in all major brain regions (12). More recently, M 5 receptors also have been detected in several peripheral tissues or cells including peripheral blood l...

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Radioligand binding assay of M1–M5 muscarinic cholinergic receptor subtypes in human peripheral blood lymphocytes

Cited by 37 publications

References 28 publications

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

An in vitro investigation of the effects of the nerve agent pretreatment pyridostigmine bromide on human peripheral blood T-cell function

Cholinergic dilation of cerebral blood vessels is abolished in M ₅ muscarinic acetylcholine receptor knockout mice

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Radioligand binding assay of M1–M5 muscarinic cholinergic receptor subtypes in human peripheral blood lymphocytes

Cited by 37 publications

References 28 publications

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

Potential clozapine target sites on peripheral hematopoietic cells and stromal cells of the bone marrow

An in vitro investigation of the effects of the nerve agent pretreatment pyridostigmine bromide on human peripheral blood T-cell function

Cholinergic dilation of cerebral blood vessels is abolished in M 5 muscarinic acetylcholine receptor knockout mice

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Cholinergic dilation of cerebral blood vessels is abolished in M ₅ muscarinic acetylcholine receptor knockout mice