Chapter 8: Autoradiographic distribution of putative muscarinic receptor sub-types in mammalian brain

doi:10.1016/S0079-6123(08)62384-5

Progress in Brain Research

Volume 98, 1993, Pages 85-93

https://doi.org/10.1016/S0079-6123(08)62384-5 Get rights and content

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The existence of multiple classes of muscarinic receptors is now well established. Major progress in that regard came about with the cloning of five different subtypes (m₁,-m₅), which are differentially regulated and expressed throughout the body. All five muscarinic receptor subtypes are members of the seven-transmembrane rhodopsin receptor superfamily and are coupled to either CAMP production or to the phosphoinositide cycle. However, little is currently known regarding the physiological role of each muscarinic receptor sub-type; their respective structure-activity relationships still being rather poorly established. It is clear that currently available radioligands lack high selectivity for a given muscarinic receptor subtype and that various complementary methodological approaches must be used, in addition to receptor autoradiography, to confirm the specific location of a given receptor class. However, in spite of this cautionary note, it is evident that multiple classes of muscarinic receptors are expressed and differentially distributed in the mammalian CNS. Further detailed anatomical information now awaits the development of tools, such as sub-type selective, high affinity M₁ to M₅ radioligands.

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Cited by (30)

Regional variation in cholinergic terminal activity determines the non-uniform occurrence of cortical slow waves during REM sleep in mice
2023, Cell Reports
Sleep consists of two basic stages: non-rapid eye movement (NREM) and rapid eye movement (REM) sleep. NREM sleep is characterized by slow high-amplitude cortical electroencephalogram (EEG) signals, while REM sleep is characterized by desynchronized cortical rhythms. Despite this, recent electrophysiological studies have suggested the presence of slow waves (SWs) in local cortical areas during REM sleep. Electrophysiological techniques, however, have been unable to resolve the regional structure of these activities because of relatively sparse sampling. Here, we map functional gradients in cortical activity during REM sleep using mesoscale imaging in mice and show local SW patterns occurring mainly in somatomotor and auditory cortical regions with minimum presence within the default mode network. The role of the cholinergic system in local desynchronization during REM sleep is also explored by calcium imaging of cholinergic activity within the cortex and analyzing structural data. We demonstrate weaker cholinergic projections and terminal activity in regions exhibiting frequent SWs during REM sleep.
Different roles for M<inf>1</inf> and M<inf>2</inf> receptors within perirhinal cortex in object recognition and discrimination
2014, Neurobiology of Learning and Memory
Citation Excerpt :
Because PRh lesions disrupt object processing in object recognition and oddity discrimination, intra-PRh infusions of M1 antagonists were predicted to impair both object recognition and oddity discrimination. Conversely, M2 receptors are located primarily presynaptically (Mrzljak, Levey, & Goldman-Rakic, 1993; Packard, Regenold, Quirion, & White, 1990) and function as autoreceptors (Galarraga et al., 1999; Quirion, Aubert, Araujo, Hersi, & Gaudreau, 1993); therefore, antagonising M2 receptors might increase cholinergic activity in PRh, possibly facilitating performance in object recognition and oddity discrimination tasks. The series of experiments examined the effects of intra-PRh infusions of pirenzepine (Experiment 1), AF-DX 116 (Experiment 2), and MT-7 (Experiment 3) in object recognition.
Recognition and discrimination of objects and individuals are critical cognitive faculties in both humans and non-human animals, and cholinergic transmission has been shown to be essential for both of these functions. In the present study we focused on the role of M₁ and M₂ muscarinic receptors in perirhinal cortex (PRh)-dependent object recognition and discrimination. The selective M₁ antagonists pirenzepine and the snake toxin MT-7, and a selective M₂ antagonist, AF-DX 116, were infused directly into PRh. Pre-sample infusions of both pirenzepine and AF-DX 116 significantly impaired object recognition memory in a delay-dependent manner. However, pirenzepine and MT-7, but not AF-DX 116, impaired oddity discrimination performance in a perceptual difficulty-dependent manner. The findings indicate distinct functions for M₁ and M₂ receptors in object recognition and discrimination.
Muscarinic receptor/G-protein coupling is reduced in the dorsomedial striatum of cognitively impaired aged rats
2012, Behavioural Brain Research
Citation Excerpt :
To measure M2/M4 receptor levels, the muscarinic receptor antagonist [3H]AFDX-398 was used. It preferentially binds to the M2 and M4 AChRs at the concentration used in this study (2 nM) ([18,20,21]). Muscarinic receptor function and levels were quantified in the same brain regions of the same rats and data were analyzed relative to both age and cognitive ability.
Behavioral flexibility, the ability to modify responses due to changing task demands, is detrimentally affected by aging with a shift towards increased cognitive rigidity. The neurobiological basis of this cognitive deficit is not clear although striatal cholinergic neurotransmission has been implicated. To investigate the possible association between striatal acetylcholine signaling with age-related changes in behavioral flexibility, young, middle-aged, and aged F344 X Brown Norway F1 rats were assessed using an attentional set-shifting task that includes two tests of behavioral flexibility: reversal learning and an extra-dimensional shift. Rats were also assessed in the Morris water maze to compare potential fronto-striatal-dependent deficits with hippocampal-dependent deficits. Behaviorally characterized rats were then assessed for acetylcholine muscarinic signaling within the striatum using oxotremorine-M-stimulated [³⁵S]GTPγS binding and [³H]AFDX-384 receptor binding autoradiography. The results showed that by old age, cognitive deficits were pronounced across cognitive domains, suggesting deterioration of both hippocampal and fronto-striatal regions. A significant decline in oxotremorine-M-stimulated [³⁵S]GTPγS binding was limited to the dorsomedial striatum of aged rats when compared to young and middle-aged rats. There was no effect of age on striatal [³H]AFDX-384 receptor binding. These results suggest that a decrease in M2/M4 muscarinic receptor coupling is involved in the age-associated decline in behavioral flexibility.
Carbachol injections into the intergeniculate leaflet induce nonphotic phase shifts
2007, Brain Research
Citation Excerpt :
The pharmacological identity of the cholinergic receptor subtypes responsible for the phase shifting effects of carbachol remains to be determined. Other studies have demonstrated that all five muscarinic acetylcholine receptors and nicotinic receptor subunits are found in the thalamus of the rat (Brann et al., 1993; Buckley et al., 1988; Levey et al., 1994; Quirion et al., 1993; Vilaro et al., 1990; Wada et al., 1989; Zubieta and Frey, 1993) and the M2 and M3 muscarinic receptors have been reported in the geniculate bodies of the rat and cat (Plummer et al., 1999). Further studies will examine the roles of the different receptor subtypes involved in mediating nonphotic phase shifting.
Nonphotic phase shifts of the circadian clock in mammals are mediated by the intergeniculate leaflet (IGL) of the thalamus via a geniculohypothalamic projection to the suprachiasmatic nucleus. These shifts can be induced by arousing stimuli, such as wheel running, brain stimulation reward and foot shock. Because mesopontine cholinergic neurons are also activated by arousing stimuli, we tested the hypothesis that cholinergic input to the IGL mediates nonphotic phase shifts. Carbachol injected into the IGL of hamsters in their subjective day (CT8) induced phase advances similar to shifts that are induced by arousal at the same circadian time. Control injections of saline at CT8 did not advance phase similarly. Carbachol injections outside the IGL produced smaller shifts. Pre-injections of the muscarinic antagonist, atropine, reduced carbachol-induced phase advances relative to saline pre-injections. The results indicate that muscarinic input to the IGL can induce nonphotic phase shifts.
Involvement of the intralaminar parafascicular nucleus in muscarinic-induced antinociception in rats
2004, Brain Research
Citation Excerpt :
However, recent studies using muscarinic receptor gene knockout mice and relatively selective muscarinic toxins suggest that M2 and M4 receptors are preferentially involved in muscarinic-induced antinociception [34,103]. Localization studies show moderate to high expression of M2 receptors in nPf [67,93], and suggest that M2 and M3 receptors are the predominant muscarinic receptor expressed in the thalamus [20,22,30,61,83,85,100,107,111]. Thus, similar to findings observed following carbachol administration into the reticular formation [64], the antinociceptive effects observed following intra-nPf carbachol are most likely mediated by M2 receptors.
The thalamic contribution to cholinergic-induced antinociception was examined by microinjecting the acetylcholine (ACh) agonist carbachol into the intralaminar nucleus parafascicularis (nPf) of rats. Pain behaviors organized at spinal (spinal motor reflexes), medullary (vocalizations during shock), and forebrain (vocalization afterdischarges, VADs) levels of the neuraxis were elicited by noxious tailshock. Carbachol (0.5, 1, and 2 μg/side) administered into nPf produced dose-dependent elevations of vocalization thresholds, but failed to elevate spinal motor reflex threshold. Injections of carbachol into adjacent sites dorsal or ventral to nPf failed to alter vocalization thresholds. Elevations in vocalization thresholds produced by intra-nPf carbachol were reversed in a dose-dependent manner by local administration of the muscarinic receptor antagonist atropine (30 and 60 μg/side). These results provide the first direct evidence supporting the involvement of the intralaminar thalamus in muscarinic-induced antinociception. Results are discussed in terms of the contribution of nPf to the processing of the affective dimension of pain.
Changes in acetylcholinesterase activity and muscarinic receptor bindings in μ-opioid receptor knockout mice
2004, Molecular Brain Research
Anatomical evidence indicates that cholinergic and opioidergic systems are co-localized and acting on the same neurons. However, the regulatory mechanisms between cholinergic and opioidergic system have not been well characterized. In the present study, we investigated whether there are compensatory changes of acetylcholinesterase activity and cholinergic receptors in mice lacking μ-opioid receptor gene. The acetylcholinesterase activity was determined by histochemistry assay. The cholinergic receptor binding was carried out by quantitative autoradiography using [³H]-quinuclidinyl benzilate (nonselective muscarinic receptors), N-[³H]-methylscopolamine (nonselective muscarinic receptors), [³H]-pirenzepine (M1 subtype muscarinic receptors) and [³H]-AF-DX384 (M2 subtype muscarinic receptors) in brain slices of wild-type and μ-opioid receptor knockout mice. The acetylcholinesterase activity of μ-opioid receptor knockout mice was higher than that of the wild-type in the striatal caudate putamen and nucleus accumbens, but not in the cortex and hippocampus areas. In addition, the bindings in N-[³H]-methylscopolamine and [³H]-AF-DX384 of μ-opioid receptor knockout mice were significantly lower when compared with that of the wild-type controls in the striatal caudate putamen and nucleus accumbens. However, there were no significant differences in bindings of [³H]-quinuclidinyl benzilate and [³H]-pirenzepine between μ-opioid receptor knockout and wild-type mice in the cortex, striatum and hippocampus. These data indicate that there are up-regulation of acetylcholinesterase activity and compensatory down-regulation of M2 muscarinic receptors in the striatal caudate putamen and nucleus accumbens of μ-opioid receptor knockout mice.

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Chapter 8: Autoradiographic distribution of putative muscarinic receptor sub-types in mammalian brain

Publisher Summary

Eur. J. Pharmacol.

Neuroscience

Brain Res.

Brain Res.

Neuroscience

Eur. J. Pharmacol.

Eur. J. Pharmacol.

Brain Res.

Eur. J. Pharmacol.

Neuroscience

Eur. J. Pharmacol.

Brain Res.

Dev. Brain Res.

Brain Res.

Prog. Neurobiol.

Differential alterations of various cholinergic markers in cortical and subcortical regions of the human brain in Alzheimer's Disease.

J. Neurochem.

Characterization of [3H]AF-DX 116 binding sites in rat brain: Evidence for heterogeneity of muscarinic-M2 receptor sites.

Synapse

Effects of aging on nicotinic and muscarinic autoreceptor function in the rat brain: Relationship to presynaptic cholinergic markers and binding sites.

J. Neurosci.

Heterogeneous binding of [3H]4-DAMP to muscarinic cholinergic sites in the rat brain: evidence from membrane binding and autoradiographic studies.

Synapse

Comparative alterations of nicotinic and muscarinic binding sites in Alzheimer's and Parkinson's Diseases.

J. Neurochem.

New subtypes of muscarinic acetylcholine receptors.

Trends Pharmacol. Sci.

The molecular basis of muscarinic receptor diversity.

Trends Neurosci.

Localization of a family of muscarinic receptor mRNAs in rat brain.

J. Neurosci.

Histochemical distribution of acetylcholinesterase in the central nervous system: clues to the localization of cholinergic neurons.

Muscarinic cholinergic receptor subtypes in the rat brain. I. Quantitative autoradiographic studies.

Brain Res.

Selectivity of muscarinic antagonists in radioligand and in vivo experiments for the putative M1, M2 and M3 receptors.

J. Pharmacol. Exp. Ther.

Antagonist binding profiles of five cloned human muscarinic receptor subtypes.

J. Pharmacol. Exp. Ther.

Labeling of rat heart muscarinic receptors using the new M2 selective antagonist [3H]AF-DX 384.

Biochem. Pharmacol.

The organization and some projections of cholinergic neurons of the mammalian forebrain.

Brain Res. Rev.

Characterization of [³H]AF-DX 116 binding sites in rat brain: Evidence for heterogeneity of muscarinic-M₂ receptor sites.

Heterogeneous binding of [³H]4-DAMP to muscarinic cholinergic sites in the rat brain: evidence from membrane binding and autoradiographic studies.

Selectivity of muscarinic antagonists in radioligand and in vivo experiments for the putative M₁, M₂ and M₃ receptors.

Labeling of rat heart muscarinic receptors using the new M₂ selective antagonist [³H]AF-DX 384.