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The Use of a Modified [3H]4-DAMP Radioligand Binding Assay With Increased Selectivity for Muscarinic M3 Receptor Shows That Cortical CHRM3 Levels Are Not Altered in Mood Disorders

The Use of a Modified [3H]4-DAMP Radioligand Binding Assay With Increased Selectivity for Muscarinic M3 Receptor Shows That Cortical CHRM3 Levels Are Not Altered in Mood Disorders

Won Je Jeon, Andrew S Gibbons, Brian Dean

Prog Neuropsychopharmacol Biol Psychiatry. 2013 Dec 2;47:7-12.

PMID: 23962466

Abstract:

[(3)H]4-DAMP is a radioligand that has been used to quantify levels of the muscarinic receptor CHRM3 protein in situ. However, in addition to high affinity binding to CHRM3, [(3)H]4-DAMP binds with low affinity to CHRM1 confounding the potential to discriminate between changes in these two muscarinic receptors. We have developed a [(3)H]4-DAMP binding assay, optimised for measuring CHRM3 protein levels in the cortex, with minimal selectivity towards CHRM1. The selectivity of our assay towards CHRM3 was confirmed using recombinant receptor-expressing, cell lysate preparations. [(3)H]4-DAMP binding levels were similar between wildtype and CHRM1 knockout mice, confirming that the amount of [(3)H]4-DAMP binding to CHRM1 was negligible. We used this assay to measure CHRM3 protein levels in the frontal pole, obtained post-mortem from subjects with bipolar disorder (n = 15), major depressive disorder (n = 15) and matched controls (n = 20) and showed that [(3)H]4-DAMP binding was not altered in either bipolar disorder or major depressive disorder. Western blotting confirmed that CHRM3 protein levels were unchanged in these subjects.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP1952154	4-DAMP		1952-15-4	Price

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