Regulation of phospholipase Cβ activity by muscarinic acetylcholine and 5-HT 2 receptors in crude and synaptosomal membranes from human cerebral cortex

Abstract

Stimulation of phospholipase Cβ by receptor agonists and G proteins has been characterized in crude cerebral membrane preparations, but little is known about their presynaptic localizations and little information is currently available for human brain tissue. The characteristics of phosphoplipase C transmembrane signaling were studied in crude and synaptosomal plasma membranes from postmortem human prefrontal cortex by measuring the hydrolysis of exogenous [ 3H]phosphatidylinositol4,5bisphosphate(PIP 2) and the immunoreactive levels of phospholipase C (PLC) and G αq/11 proteins. Regulation of PLC activity by Ca 2+ and the 5-HT 2 receptor agonist 5-methyltryptamine, but not by guanosine 5′- O-[3-thiotriphosphate] and the muscarinic acetylcholine receptor agonist carbachol were different between crude and synaptosomal membranes. KCl (20 mM) stimulation was absent in both preparations. Levels of G αq/11-protein subunits differed between preparations. The functional inhibition carried out with pirenzepine in crude membranes in order to reverse the carbachol-induced PLC stimulation indicates the existence of a component (53%) of the response that is activated by the M 1 muscarinic acetylcholine receptor subtype, and another component (47%) probably mediated by the M 3 muscarinic acetylcholine receptor subtype. In synaptosomal plasma membranes an increased inhibition of carbachol-induced PLC activation through M 1 was found. The PLC activation by 5-methyltryptamine (ketanserin-sensitive in crude membranes) was absent in synaptosomal plasma membranes suggesting the lack of activity mediated by 5-HT 2-serotonin receptors.