Regulation and function of cyclic GMP-mediated pathways in glial cells.

A large body of evidence supports a role for the NO-cGMP-protein kinase G pathway in the regulation of synaptic transmission and plasticity, brain development and neuroprotection. Circumstantial evidence implicates natriuretic peptide-stimulated cGMP formation in the same CNS functions. In addition to neurons, both cGMP-mediated pathways are functional in glial cells and an increasing number of reports indicate that they may control important aspects of glial cell physiology relevant to neuronal function. In this article we briefly review the regulation of cGMP formation in glial cells and summarize recent evidence indicating that cGMP-mediated pathways can play important roles in astroglial and microglial function in normal and diseased brain.

The ANP-cGMP-protein kinase G pathway induces a phagocytic phenotype but decreases inflammatory gene expression in microglial cells.

Reactive gliosis is a prominent feature of CNS injury that involves dramatic changes in glial cell morphology together with increased motility, phagocytic activity, and release of inflammatory mediators. We have recently demonstrated that stimulation of the cGMP-protein kinase G (PKG) pathway by NO or atrial natriuretic peptide (ANP) regulates cytoskeleton dynamics and motility in rat astrocytes in culture. In this work, we show that the cGMP-PKG pathway stimulated by ANP, but not by NO, regulates microglial cell morphology by inducing a dramatic reorganization in the actin cytoskeleton. Both ANP (0.01-1.0 microM) and the permeable cGMP analog, dibutyryl-cGMP (1-100 microM), promote a rapid (maximal at 30 min) and concentration-dependent increase in size, rounding, and lamellipodia and filopodia formation in rat brain cultured microglia. These morphological changes involve an augment and redistribution of F-actin and result in increased phagocytic activity. ANP-induced rearrangements in actin cytoskeleton and inert particle phagocytosis are prevented by the PKG inhibitor, Rp-8-Br-PET-cGMPS (0.5 microM), and involve inhibition of RhoA GTPase and activation of Rac1 and Cdc42. However, ANP does not induce NO synthase Type 2 (NOS-2) or tumor necrosis factor-alpha expression and is able to decrease lipopolysaccharide (LPS)-elicited induction of these inflammatory genes. The morphological changes and the decrease of LPS-induced NOS-2 expression produced by ANP in cultured microglia are also observed by immunostaining in organotypic cultures from rat hippocampus. These results suggest that stimulation of the ANP-cGMP-PKG pathway in microglia could play a beneficial role in the resolution of neuroinflammation by removing dead cells and decreasing levels of proinflammatory mediators.

The cyclic GMP-protein kinase G pathway regulates cytoskeleton dynamics and motility in astrocytes.

We have previously demonstrated that inflammatory compounds that increase nitric oxide (NO) synthase expression have a biphasic effect on the level of the NO messenger cGMP in astrocytes. In this work, we demonstrate that NO-dependent cGMP formation is involved in the morphological change induced by lipopolysaccharide (LPS) in cultured rat cerebellar astroglia. In agreement with this, dibutyryl-cGMP, a permeable cGMP analogue, and atrial natriuretic peptide, a ligand for particulate guanylyl cyclase, are both able to induce process elongation and branching in astrocytes resulting from a rapid, reversible and concentration-dependent redistribution of glial fibrillary acidic protein (GFAP) and actin filaments without significant change in protein levels. These effects are also observed in astrocytes co-cultured with neurons. The cytoskeleton rearrangement induced by cGMP is prevented by the specific protein kinase G inhibitor Rp-8Br-PET-cGMPS and involves downstream inhibition of RhoA GTPase since is not observed in cells transfected with constitutively active RhoA. Furthermore, dibutyryl-cGMP prevents RhoA-membrane association, a step necessary for its interaction with effectors. Stimulation of the cGMP-protein kinase G pathway also leads to increased astrocyte migration in an in vitro scratch-wound assay resulting in accelerated wound closure, as seen in reactive gliosis following brain injury. These results indicate that cGMP-mediated pathways may regulate physio-pathologically relevant responses in astroglial cells.