Effects of amyloid peptides on cell viability and expression of neuropeptides in cultured rat dorsal root ganglion neurons: a role for free radicals and protein kinase C.

Abstract

Chronic pain caused by nerve injury and inflammation is more common in the elderly. However, mechanisms underlying this phenomenon are unclear. Higher sensitivity of sensory neurons to free radicals has been suggested as one possibility. The production of free radicals can be induced by various agents, including the highly toxic protein beta-amyloid (A beta), which is found in higher amounts in the brains of Alzheimer's Disease patients. In dorsal root ganglion (DRG) cultures exposed to A beta, we examined cellular toxicity and peptide expression, in particular calcitonin gene-related peptide (CGRP), a peptide which is abundantly expressed by nociceptive afferents and is known to be involved in pain processes. Exposure of cultured rat DRG neurons to A beta(25--35) or A beta(1--40) (10 or 20 microM for 24--96 h) increased trypan blue-stained cells in a concentration- and time-dependent manner, thus, indicating cellular toxicity. These treatments also increased the number of CGRP immunoreactive (IR) neurons while decreasing the number of neuropeptide Y- and galanin-IR neurons. The free radical scavenger, superoxide dismutase, attenuated both the toxicity and neuropeptide changes induced by A beta, thus, suggesting that oxidative stress probably contributes to these effects. Exposure of cultured DRG neurons to A beta also increased the number of protein kinase C alpha (PKC alpha)-IR neurons. The PKC inhibitors, chelerythrine chloride and Gö6976, significantly augmented A beta-induced cellular toxicity while attenuating the increases in CGRP-and PKC alpha-IR cells, supporting the notion of a protective role for PKC in A beta insults. These in vitro data suggest that A beta peptides may, in addition to causing neurotoxicity, regulate neuropeptide expression in primary afferents. This finding could be relevant to the higher incidence of neuropathic pain that occurs with ageing.