Muscarinic receptor-activated signal transduction pathways involved in the neuritogenic effect of astrocytes in hippocampal neurons

Astrocytes have been shown to release factors that affect various aspects of neuronal development. We have previously shown that the acetylcholine analog carbachol, by activating muscarinic M₃ receptors in rat astrocytes, increases their ability to promote neuritogenesis in hippocampal neurons. This effect was mediated by an increased expression and release by astrocytes of several permissive factors, a most relevant of which was fibronectin. In the present study we investigated the signal transduction pathways involved in these effects of carbachol in astrocytes. Results show that multiple pathways are involved in the effects of carbachol on astrocyte-mediated increases in fibronectin expression and neuritogenesis. These include the phospholipase D pathway, leading to sequential activation of protein kinase C (PKC) ζ, p70S6 kinase and nuclear factor-κB; the phosphoinositide-3 kinase pathway; and the PKC ε pathway leading to activation of mitogen activated protein kinase. These pathways were shown to mediate the effect of carbachol on neurite outgrowth as well as the increased expression of fibronectin, further substantiating the important role of the latter in astrocyte-mediated neuritogenesis. Interference with these signaling pathways would be expected to impair astrocyte-neurons communication leading to impaired neuronal development.