Muscarinic cholinergic receptors subtypes in rat cerebellar cortex: light microscope autoradiography of age-related changes

doi:10.1016/S0006-8993(00)03146-2

Brain Research

Volume 889, Issues 1–2, 19 January 2001, Pages 256-259

https://doi.org/10.1016/S0006-8993(00)03146-2 Get rights and content

Abstract

Muscarinic cholinergic M1–M5 receptor subtypes were investigated in the cerebellar cortex of Fischer 344 rats aged 6 (young), 15 (adult) and 22 months (senescent) by combined kinetic and equilibrium binding and light microscope autoradiography. In young rats the rank order of receptor density was M5<M4<M3 and M3<M5<M4 in the molecular and granular layers, respectively. M1, M2, M4 and M5 receptors were also observed within Purkinje neurons. M1 receptor did not show age-related changes as well as the M2 receptor in the molecular layer. In this layer, M3–M5 receptors were increased in senescent compared to younger rats. In the granular layer the expression of M2 and M5 muscarinic receptors was similar in young and senescent rats and higher in adult rats. M3 and M4 receptors were more in adult and senescent rats compared to young animals. In Purkinje neurons, a slight-to-moderate age-related increase of M1 and M5 receptor expression was observed.

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Citation Excerpt :
For labeling M5 muscarinic cholinergic receptor, sections were pre-incubated in a phosphate-MgCl2 buffer containing 30 mg/ml of Mamba venom for 30 min at 20 °C, and then incubated with 1 μM AQ-RA 741 and 0.5 nM [3H]NMS for 30 min at 20 °C. Sections were then transferred in scintillation vials that were read in a liquid scintillation spectrometer or processed for high resolution (using nuclear emulsion-coated cover slips to reveal radioactivity) light microscope autoradiography according to the procedure detailed in a previous study of our group (Tayebati et al., 2001). For assessing levels of non-specific retention of radioligand by sections, some slides were incubated for 60 min at 20 °C with 0.5 nM [3H]NMS alone (total binding) or plus 1 μM atropine (non-specific binding).
Striatum expresses a cholinergic system involved in the regulation of its activity and changes in striatal cholinergic receptors may be related to cognitive impairment. This study has investigated muscarinic cholinergic M1–M5 receptor subtype expression in striatum of Fischer 344 rats aged 6 (young), 15 (adult) and 22 months (senescent) to assess the contribution of different muscarinic cholinergic receptor subtypes in age-related changes of striatal cholinergic neurotransmission. Western blot analysis revealed the expression of the M1–M5 muscarinic receptor subtytpes in the striatum of rats of the three age groups investigated. Both radioligand binding assay and light microscope autoradiography showed in young rats a M4>M1>M2>M3>M5 rank order of receptor density. With the exception of M1 receptor, the density of which is similar in the dorsal (motor) and ventral (limbic) striatum, other receptor subtypes were more abundant in ventral than in dorsal striatum. M1 receptor expression was unchanged between young and adult rats and decreased in senescent animals both in dorsal and ventral striatum. In dorsal striatum M2 and M5 receptor expression did not show age-related changes, whereas in ventral striatum it was slightly decreased in adult rats compared to young or senescent cohorts. M3 receptor expression did not show age-related modifications, whereas a progressive age-related decrease of M4 receptor was found, both in dorsal and ventral striatum. These data indicate a heterogeneous response to age of different muscarinic receptor subtypes. Striatal cholinergic markers are thought to correlate with cognitive impairment in aged rats. In view of this, the identification of age-related changes of striatal muscarinic receptor subtypes may contribute to develop cholinergic strategies to counter cholinergic neurotransmission changes occurring with aging.
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Short communicationMuscarinic cholinergic receptors subtypes in rat cerebellar cortex: light microscope autoradiography of age-related changes

Abstract

Pharmacol. Ther.

Prog. Brain Res.

Anat. Anz.

Neuropharmacology

Life Sci.

Brain Res.

J. Neuroimmunol.

Short communication
Muscarinic cholinergic receptors subtypes in rat cerebellar cortex: light microscope autoradiography of age-related changes