Muscarinic Acetylcholine Receptors Induce the Expression of the Immediate Early Growth Regulatory Gene CYR61

Abstract

In brain, muscarinic acetylcholine receptors (mAChRs) modulate neuronal functions including long term potentiation and synaptic plasticity in neuronal circuits that are involved in learning and memory formation. To identify mAChR-inducible genes, we used a differential display approach and found that mAChRs rapidly induced transcription of the immediate early gene CYR61 in HEK 293 cells with a maximum expression after 1 h of receptor stimulation. CYR61 is a member of the emerging CCN gene family that includes CYR61/CEF10, CTGF/FISP-12, and NOV; these encode secretory growth regulatory proteins with distinct functions in cell proliferation, migration, adhesion, and survival. We found that CYR61, CTGF, and NOV were expressed throughout the human central nervous system. Stimulation of mAChRs induced CYR61 expression in primary neurons and rat brain where CYR61 mRNA was detected in cortical layers V and VI and in thalamic nuclei. In contrast, CTGF and NOV expression was not altered by mAChRs neither in neuronal tissue culture nor rat brain. Receptor subtype analyses demonstrated that m1 and m3 mAChR subtypes strongly induced CYR61 expression, whereas m2 and m4 mAChRs had only subtle effects. Increased CYR61 expression was coupled to mAChRs by both protein kinase C and elevations of intracellular Ca 2؉ . Our results establish that CYR61 expression in mammalian brain is under the control of cholinergic neurotransmission; it may thus be involved in cholinergic regulation of synaptic plasticity.

The C-terminal Tail of the M3-muscarinic Receptor Possesses Anti-apoptotic Properties

“…This suggested that the mechanism of the M 3 -muscarinic receptor response was rapid in its onset and was then maintained for an extended period after agonist withdrawal. A strong candidate for mediating such a mechanism was rapid changes in intracellular calcium, which in the case of a number of GPCRs has been shown to encode for longer adaptive responses such as changes in gene transcription (24,25). However, analysis of a truncated mutant of the M 3 -muscarinic receptor (⌬565-M 3 ), which was unable to mediate an anti-apoptotic response, revealed that this receptor was still coupled to the calcium/PLC pathway in a manner similar to the wild-type receptor.…”

Expression and regulation of Cyr61 in human breast cancer cell lines

“…Phorbol ester 12-O-tetradecanoyphorbol-13-acetate (TPA) mediates Cyr61 expression in the liver (Nathans et al, 1988). Incidentally, others and we have found expression of Cyr61 involving protein kinase C (PKC) and the MEK/MAPK pathways (Chung and Ahn, 1998;Inuzuka et al, 1999;Albrecht et al, 2000;Tsai et al, 2001, manuscript submitted). Cyr61 is also vitamin D-responsive in human osteoblasts and osteosarcoma cell lines (Scheutze et al, 1998).…”

“…Other cytokines, such as tumor necrosis factor a (TNFa) and interleukin-1 (IL-1) also strongly elevate the expression of Cyr61 (Scheutze et al, 1998). Cyr61 is di erentially expressed by muscarinic acetylcholine receptors (mAChRs) in the brain (Albrecht et al, 2000), and by factor VIIa and thrombin in human ®broblasts (Pendurthi et al, 2000). Phorbol ester 12-O-tetradecanoyphorbol-13-acetate (TPA) mediates Cyr61 expression in the liver (Nathans et al, 1988).…”

Mechanisms involved in the regulation of mRNA for M2 muscarinic acetylcholine receptors and endothelial and neuronal NO synthases in rat atria

“…Within 1 h of M 1 receptor activation, the expression of immediate early genes is induced. These genes play important roles in coupling receptor stimulation to long-term neuronal responses (Albrecht et al, 2000). Three hours after M 1 receptor activation, a significantly higher number of genes, some expressing differential patterns, were observed.…”