cAMP promotes neurite outgrowth and extension through protein kinase A but independently of Erk activation in cultured rat motoneurons.

It is well established that cAMP counteracts myelin inhibition to permit axon regeneration in the central nervous system. On the other hand, the role of cAMP in axonal growth on permissive substrates remains controversial because the evidence available is contradictory. In view that elevation of cAMP represents an attractive therapeutic target to promote nerve regeneration in vivo, we investigated the effect of cAMP on neurite outgrowth and extension in motoneurons. We manipulated cAMP levels pharmacologically in cultured motoneurons and investigated targets downstream of cAMP of neurite outgrowth and extension on a permissive substrate. Reduction of cAMP by the adenylyl cyclase inhibitor SQ22536 inhibited, and elevation of cAMP by forskolin, dibutyryl cAMP, IBMX and rolipram increased outgrowth and extension of neurites. The cAMP-mediated effects occur via activation of protein kinase A (PKA) and were reduced by the inhibitors, H89 and Rp-cAMP. However, cAMP elevation did not lead to Erk activation that is an essential downstream component of neurotrophin signaling. These findings provide evidence for a key role of cAMP in promoting peripheral nerve regeneration after nerve injuries and indicate that this effect is unusual in not being mediated via Erk phosphorylation.

Uptake of lipoproteins for axonal growth of sympathetic neurons.

Lipoproteins originating from axon and myelin breakdown in injured peripheral nerves are believed to supply cholesterol to regenerating axons. We have used compartmented cultures of rat sympathetic neurons to investigate the utilization of lipids from lipoproteins for axon elongation. Lipids and proteins from human low density lipoproteins (LDL) and high density lipoproteins (HDL) were taken up by distal axons and transported to cell bodies, whereas cell bodies/proximal axons internalized these components from only LDL, not HDL. Consistent with these observations, the impairment of axonal growth, induced by inhibition of cholesterol synthesis, was reversed when LDL or HDL were added to distal axons or when LDL, but not HDL, were added to cell bodies. LDL receptors (LDLRs) and LR7/8B (apoER2) were present in cell bodies/proximal axons and distal axons, with LDLRs being more abundant in the former. Inhibition of cholesterol biosynthesis increased LDLR expression in cell bodies/proximal axons but not distal axons. LR11 (SorLA) was restricted to cell bodies/proximal axons and was undetectable in distal axons. Neither the LDL receptor-related protein nor the HDL receptor, SR-B1, was detected in sympathetic neurons. These studies demonstrate for the first time that lipids are taken up from lipoproteins by sympathetic neurons for use in axonal regeneration.